Bio

Academic Appointments


Honors & Awards


  • Member, National Academy of Sciences (2012)
  • Fellow, American Academy of Arts and Sciences (2012)
  • Fellow, American Association for the Advancement of Science (2011)
  • H.W.Mossman Award, American Association of Anatomists (2007)
  • Jacob Javits Award, National Institute of Neurological Disorders and Stroke (2005)
  • Investigator, Howard Hughes Medical Institute (2005)
  • Young Investigator Award, Society for Neuroscience (2002)
  • Technology Innovation Award in Neuroscience, McKnight Foundation (2002)

Professional Education


  • B.S., Univ. of Sci. & Tech. of China, Molecular Biology (1986)
  • Ph.D., Brandeis University, Biology (1992)

Research & Scholarship

Current Research and Scholarly Interests


1. Organization of the olfactory system
The olfactory systems from flies to mammals use a similar organizational principle. Olfactory receptor neurons (ORNs) expressing the same odorant receptor project their axons to the same glomerulus. Projection neurons (PNs) send dendrites to individual glomeruli, and relay olfactory information via their axons to high olfactory centers. Using MARCM (see below) to label individual fly PNs, we found that PN axon terminals exhibit striking stereotypy at the lateral horn according to the glomeruli they send dendrites to. Axon terminals of PNs representing food odors are spatially segregated from those that represent mating pheromones. By contrast, PN axon terminal arborizations in the mushroom body, the olfactory learning and memory center, exhibit much less stereotypy. We are currently using two-photon calcium imaging, optogenetics, and quantitative behavioral assays to identify principles of information processing at the antennal lobe and in higher olfactory centers suggested by previous anatomical studies. We are also investigating how the glomerular map in the mouse olfactory bulb is represented in olfactory cortex using virus-mediated trans-synaptic tracing.

2. Development of wiring specificity in the fly olfactory system
The assembly of the fly olfactory system requires precise glomerular targeting of axons from each of the 50 ORN classes, as well as dendrites of each of the 50 PN classes. We are using this neural circuit as a model to investigate the general principles by which precise wiring specificity arises during development. Our previous studies have shown that PN dendrite patterning precedes ORN axon targeting. PN dendrite targeting relies on global cues in the form of gradients, as well as local cues distributed in a “salt-and-pepper” fashion on dendrites projecting to different glomeruli. Targeting of ORN axons may use the same molecules as PN dendrite targeting, but via distinct mechanisms including axon-axon interactions and axon-target interactions. We are currently performing systematic genetic studies to identify the cell-surface code ORNs and PNs use to form specific connections at stereotypically organized glomeruli.

3. Developmental neurobiology
In addition to our focus on the olfactory system, we are investigating several other developmental neurobiological problems. These include mechanisms of axon pruning, the roles of neuronal activity in neuronal maturation and incorporation into functional circuits, and cell autonomous functions of genes that are implicated in human neurological disorders. We are using both fly and mouse systems to study these problems.

4. Creating genetic tools
In the process of dissecting the adult organization and developmental assembly of complex neural circuits, we have created several useful genetic tools. The MARCM method (Mosaic Analysis with a Repressible Cell Marker) enables the visualization and genetic manipulation of small populations of cells or single neurons in a mosaic fly. We have developed a new repressible binary expression system, the Q system, which has many applications and is helping us to study several problems described above.
We have also developed a mosaic method in the mouse called MADM (Mosaic Analysis with Double Markers) that allows sparse labeling and genetic manipulation of individual cells or cells that share the same lineage with distinct colors in mosaic animals. We have used MADM as trace lineages and study cell autonomous gene functions in neural developmental processes. We are currently expanding the MADM technique to other mouse chromosomes, and will use MADM to study several developmental neurobiological problems (see above). We have also developed other useful mice, such as a double fluorescent Cre reporter and synapse labeling tools in vivo.

Teaching

2013-14 Courses


Graduate and Fellowship Programs


Publications

Journal Articles


  • Permanent Genetic Access to Transiently Active Neurons via TRAP: Targeted Recombination in Active Populations. Neuron Guenthner, C. J., Miyamichi, K., Yang, H. H., Heller, H. C., Luo, L. 2013; 78 (5): 773-784

    Abstract

    Targeting genetically encoded tools for neural circuit dissection to relevant cellular populations is a major challenge in neurobiology. We developed an approach, targeted recombination in active populations (TRAP), to obtain genetic access to neurons that were activated by defined stimuli. This method utilizes mice in which the tamoxifen-dependent recombinase CreER(T2) is expressed in an activity-dependent manner from the loci of the immediate early genes Arc and Fos. Active cells that express CreER(T2) can only undergo recombination when tamoxifen is present, allowing genetic access to neurons that are active during a time window of less than 12 hr. We show that TRAP can provide selective access to neurons activated by specific somatosensory, visual, and auditory stimuli and by experience in a novel environment. When combined with tools for labeling, tracing, recording, and manipulating neurons, TRAP offers a powerful approach for understanding how the brain processes information and generates behavior.

    View details for DOI 10.1016/j.neuron.2013.03.025

    View details for PubMedID 23764283

  • Linking cell fate, trajectory choice, and target selection: genetic analysis of sema-2b in olfactory axon targeting. Neuron Joo, W. J., Sweeney, L. B., Liang, L., Luo, L. 2013; 78 (4): 673-686

    Abstract

    Neural circuit assembly requires selection of specific cell fates, axonal trajectories, and synaptic targets. By analyzing the function of a secreted semaphorin, Sema-2b, in Drosophila olfactory receptor neuron (ORN) development, we identified multiple molecular and cellular mechanisms that link these events. Notch signaling limits Sema-2b expression to ventromedial ORN classes, within which Sema-2b cell-autonomously sensitizes ORN axons to external semaphorins. Central-brain-derived Sema-2a and Sema-2b attract Sema-2b-expressing axons to the ventromedial trajectory. In addition, Sema-2b/PlexB-mediated axon-axon interactions consolidate this trajectory choice and promote ventromedial axon-bundle formation. Selecting the correct developmental trajectory is ultimately essential for proper target choice. These findings demonstrate that Sema-2b couples ORN axon guidance to postsynaptic target neuron dendrite patterning well before the final target selection phase, and exemplify how a single guidance molecule can drive consecutive stages of neural circuit assembly with the help of sophisticated spatial and temporal regulation.

    View details for DOI 10.1016/j.neuron.2013.03.022

    View details for PubMedID 23719164

  • Teneurins instruct synaptic partner matching in an olfactory map NATURE Hong, W., Mosca, T. J., Luo, L. 2012; 484 (7393): 201-U82

    Abstract

    Neurons are interconnected with extraordinary precision to assemble a functional nervous system. Compared to axon guidance, far less is understood about how individual pre- and postsynaptic partners are matched. To ensure the proper relay of olfactory information in the fruitfly Drosophila, axons of ?50 classes of olfactory receptor neurons (ORNs) form one-to-one connections with dendrites of ?50 classes of projection neurons (PNs). Here, using genetic screens, we identified two evolutionarily conserved, epidermal growth factor (EGF)-repeat containing transmembrane Teneurin proteins, Ten-m and Ten-a, as synaptic-partner-matching molecules between PN dendrites and ORN axons. Ten-m and Ten-a are highly expressed in select PN-ORN matching pairs. Teneurin loss- and gain-of-function cause specific mismatching of select ORNs and PNs. Finally, Teneurins promote homophilic interactions in vitro, and Ten-m co-expression in non-partner PNs and ORNs promotes their ectopic connections in vivo. We propose that Teneurins instruct matching specificity between synaptic partners through homophilic attraction.

    View details for DOI 10.1038/nature10926

    View details for Web of Science ID 000303149900027

    View details for PubMedID 22425994

  • Trans-synaptic Teneurin signalling in neuromuscular synapse organization and target choice NATURE Mosca, T. J., Hong, W., Dani, V. S., Favaloro, V., Luo, L. 2012; 484 (7393): 237-U122

    Abstract

    Synapse assembly requires trans-synaptic signals between the pre- and postsynapse, but our understanding of the essential organizational molecules involved in this process remains incomplete. Teneurin proteins are conserved, epidermal growth factor (EGF)-repeat-containing transmembrane proteins with large extracellular domains. Here we show that two Drosophila Teneurins, Ten-m and Ten-a, are required for neuromuscular synapse organization and target selection. Ten-a is presynaptic whereas Ten-m is mostly postsynaptic; neuronal Ten-a and muscle Ten-m form a complex in vivo. Pre- or postsynaptic Teneurin perturbations cause severe synapse loss and impair many facets of organization trans-synaptically and cell autonomously. These include defects in active zone apposition, release sites, membrane and vesicle organization, and synaptic transmission. Moreover, the presynaptic microtubule and postsynaptic spectrin cytoskeletons are severely disrupted, suggesting a mechanism whereby Teneurins organize the cytoskeleton, which in turn affects other aspects of synapse development. Supporting this, Ten-m physically interacts with ?-Spectrin. Genetic analyses of teneurin and neuroligin reveal that they have differential roles that synergize to promote synapse assembly. Finally, at elevated endogenous levels, Ten-m regulates target selection between specific motor neurons and muscles. Our study identifies the Teneurins as a key bi-directional trans-synaptic signal involved in general synapse organization, and demonstrates that proteins such as these can also regulate target selection.

    View details for DOI 10.1038/nature10923

    View details for Web of Science ID 000303149900034

    View details for PubMedID 22426000

  • Cortical representations of olfactory input by trans-synaptic tracing NATURE Miyamichi, K., Amat, F., Moussavi, F., Wang, C., Wickersham, I., Wall, N. R., Taniguchi, H., Tasic, B., Huang, Z. J., He, Z., Callaway, E. M., Horowitz, M. A., Luo, L. 2011; 472 (7342): 191-196

    Abstract

    In the mouse, each class of olfactory receptor neurons expressing a given odorant receptor has convergent axonal projections to two specific glomeruli in the olfactory bulb, thereby creating an odour map. However, it is unclear how this map is represented in the olfactory cortex. Here we combine rabies-virus-dependent retrograde mono-trans-synaptic labelling with genetics to control the location, number and type of 'starter' cortical neurons, from which we trace their presynaptic neurons. We find that individual cortical neurons receive input from multiple mitral cells representing broadly distributed glomeruli. Different cortical areas represent the olfactory bulb input differently. For example, the cortical amygdala preferentially receives dorsal olfactory bulb input, whereas the piriform cortex samples the whole olfactory bulb without obvious bias. These differences probably reflect different functions of these cortical areas in mediating innate odour preference or associative memory. The trans-synaptic labelling method described here should be widely applicable to mapping connections throughout the mouse nervous system.

    View details for DOI 10.1038/nature09714

    View details for Web of Science ID 000289469100036

    View details for PubMedID 21179085

  • Mosaic Analysis with Double Markers (MADM) in Mice. Cold Spring Harbor protocols Espinosa, J. S., Tea, J. S., Luo, L. 2014; 2014 (2)

    Abstract

    The human brain comprises more than 100 billion neurons, each of which has an elaborate shape and a complex pattern of connections. To untangle this complexity, it is often useful to visualize one neuron at a time. Mosaic analysis with double markers (MADM) is a genetic method for labeling and manipulating individual neurons. This method was developed in mice and it allows simultaneous labeling and gene knockout in clones of somatic cells or isolated single cells in vivo. In MADM, labeling is achieved by using site-specific recombinases to induce the reconstitution of chimeric fluorescent proteins. Here we provide the standard procedure for utilizing MADM to examine lineage analysis, neural circuit tracing, and gene function. ROSA26-MADM is used as an example because the reagents are published and available. As MADM cassettes are introduced onto more chromosomes, genes located on these other chromosomes can be subjected to mosaic analysis in an analogous manner to that described below. We present detailed protocols with troubleshooting guides, as well as applications of the technique in tracing neural circuits, live imaging of developing neurons, and studying mechanisms of neuronal morphogenesis.

    View details for DOI 10.1101/pdb.prot080366

    View details for PubMedID 24492775

  • Specific Kinematics and Motor-Related Neurons for Aversive Chemotaxis in Drosophila CURRENT BIOLOGY Gao, X. J., Potter, C. J., Gohl, D. M., Silies, M., Katsov, A. Y., Clandinin, T. R., Luo, L. 2013; 23 (13): 1163-1172

    Abstract

    Chemotaxis, the ability to direct movements according to chemical cues in the environment, is important for the survival of most organisms. The vinegar fly, Drosophila melanogaster, displays robust olfactory aversion and attraction, but how these behaviors are executed via changes in locomotion remains poorly understood. In particular, it is not clear whether aversion and attraction bidirectionally modulate a shared circuit or recruit distinct circuits for execution.Using a quantitative behavioral assay, we determined that both aversive and attractive odorants modulate the initiation and direction of turns but display distinct kinematics. Using genetic tools to perturb these behaviors, we identified specific populations of neurons required for aversion, but not for attraction. Inactivation of these populations of cells affected the completion of aversive turns, but not their initiation. Optogenetic activation of the same populations of cells triggered a locomotion pattern resembling aversive turns. Perturbations in both the ellipsoid body and the ventral nerve cord, two regions involved in motor control, resulted in defects in aversion.Aversive chemotaxis in vinegar flies triggers ethologically appropriate kinematics distinct from those of attractive chemotaxis and requires specific motor-related neurons.

    View details for DOI 10.1016/j.cub.2013.05.008

    View details for Web of Science ID 000321605600017

    View details for PubMedID 23770185

  • Plum, an Immunoglobulin Superfamily Protein, Regulates Axon Pruning by Facilitating TGF-beta Signaling NEURON Yu, X. M., Gutman, I., Mosca, T. J., Iram, T., Oezkan, E., Garcia, K. C., Luo, L., Schuldiner, O. 2013; 78 (3): 456-468

    Abstract

    Axon pruning during development is essential for proper wiring of the mature nervous system, but its regulation remains poorly understood. We have identified an immunoglobulin superfamily (IgSF) transmembrane protein, Plum, that is cell autonomously required for axon pruning of mushroom body (MB) γ neurons and for ectopic synapse refinement at the developing neuromuscular junction in Drosophila. Plum promotes MB γ neuron axon pruning by regulating the expression of Ecdysone Receptor-B1, a key initiator of axon pruning. Genetic analyses indicate that Plum acts to facilitate signaling of Myoglianin, a glial-derived TGF-β, on MB γ neurons upstream of the type-I TGF-β receptor Baboon. Myoglianin, Baboon, and Ecdysone Receptor-B1 are also required for neuromuscular junction ectopic synapse refinement. Our study highlights both IgSF proteins and TGF-β facilitation as key promoters of developmental axon elimination and demonstrates a mechanistic conservation between MB axon pruning during metamorphosis and the refinement of ectopic larval neuromuscular connections.

    View details for DOI 10.1016/j.neuron.2013.03.004

    View details for Web of Science ID 000318961700008

  • Mosaic Analysis with Double Markers Reveals Cell-Type-Specific Paternal Growth Dominance CELL REPORTS Hippenmeyer, S., Johnson, R. L., Luo, L. 2013; 3 (3): 960-967

    Abstract

    Genomic imprinting leads to preferred expression of either the maternal or paternal alleles of a subset of genes. Imprinting is essential for mammalian development, and its deregulation causes many diseases. However, the functional relevance of imprinting at the cellular level is poorly understood for most imprinted genes. We used mosaic analysis with double markers (MADM) in mice to create uniparental disomies (UPDs) and to visualize imprinting effects with single-cell resolution. Although chromosome 12 UPD did not produce detectable phenotypes, chromosome 7 UPD caused highly significant paternal growth dominance in the liver and lung, but not in the brain or heart. A single gene on chromosome 7, encoding the secreted insulin-like growth factor 2 (IGF2), accounts for most of the paternal dominance effect. Mosaic analyses implied additional imprinted loci on chromosome 7 acting cell autonomously to transmit the IGF2 signal. Our study reveals chromosome- and cell-type specificity of genomic imprinting effects.

    View details for DOI 10.1016/j.celrep.2013.02.002

    View details for Web of Science ID 000321896000036

    View details for PubMedID 23453967

  • Neuroscience. dSarm-ing axon degeneration. Science Yu, X. M., Luo, L. 2012; 337 (6093): 418-419

    View details for DOI 10.1126/science.1226150

    View details for PubMedID 22837513

  • The SUMO Protease Verloren Regulates Dendrite and Axon Targeting in Olfactory Projection Neurons JOURNAL OF NEUROSCIENCE Berdnik, D., Favaloro, V., Luo, L. 2012; 32 (24): 8331-8340

    Abstract

    Sumoylation is a post-translational modification regulating numerous biological processes. Small ubiquitin-like modifier (SUMO) proteases are required for the maturation and deconjugation of SUMO proteins, thereby either promoting or reverting sumoylation to modify protein function. Here, we show a novel role for a predicted SUMO protease, Verloren (Velo), during projection neuron (PN) target selection in the Drosophila olfactory system. PNs target their dendrites to specific glomeruli within the antennal lobe (AL) and their axons stereotypically into higher brain centers. We uncovered mutations in velo that disrupt PN targeting specificity. PN dendrites that normally target to a particular dorsolateral glomerulus instead mistarget to incorrect glomeruli within the AL or to brain regions outside the AL. velo mutant axons also display defects in arborization. These phenotypes are rescued by postmitotic expression of Velo in PNs but not by a catalytic domain mutant of Velo. Two other SUMO proteases, DmUlp1 and CG12717, can partially compensate for the function of Velo in PN dendrite targeting. Additionally, mutations in SUMO and lesswright (which encodes a SUMO conjugating enzyme) similarly disrupt PN targeting, confirming that sumoylation is required for neuronal target selection. Finally, genetic interaction studies suggest that Velo acts in SUMO deconjugation rather than in maturation. Our study provides the first in vivo evidence for a specific role of a SUMO protease during neuronal target selection that can be dissociated from its functions in neuronal proliferation and survival.

    View details for DOI 10.1523/JNEUROSCI.6574-10.2012

    View details for Web of Science ID 000305295600024

    View details for PubMedID 22699913

  • Controlling gene expression with the Q repressible binary expression system in Caenorhabditis elegans NATURE METHODS Wei, X., Potter, C. J., Luo, L., Shen, K. 2012; 9 (4): 391-U105

    Abstract

    We established a transcription-based binary gene expression system in Caenorhabditis elegans using the recently developed Q system. This system, derived from genes in Neurospora crassa, uses the transcriptional activator QF to induce the expression of target genes. Activation can be efficiently suppressed by the transcriptional repressor QS, and suppression can be relieved by the nontoxic small molecule quinic acid. We used QF, QS and quinic acid to achieve temporal and spatial control of transgene expression in various tissues in C. elegans. We also developed a split Q system, in which we separated QF into two parts encoding its DNA-binding and transcription-activation domains. Each domain showed negligible transcriptional activity when expressed alone, but expression of both reconstituted QF activity, providing additional combinatorial power to control gene expression.

    View details for DOI 10.1038/NMETH.1929

    View details for Web of Science ID 000302218500024

    View details for PubMedID 22406855

  • Extensions of MADM (Mosaic Analysis with Double Markers) in Mice PLOS ONE Tasic, B., Miyamichi, K., Hippenmeyer, S., Dani, V. S., Zeng, H., Joo, W., Zong, H., Chen-Tsai, Y., Luo, L. 2012; 7 (3)

    Abstract

    Mosaic Analysis with Double Markers (MADM) is a method for generating genetically mosaic mice, in which sibling mutant and wild-type cells are labeled with different fluorescent markers. It is a powerful tool that enables analysis of gene function at the single cell level in vivo. It requires transgenic cassettes to be located between the centromere and the mutation in the gene of interest on the same chromosome. Here we compare procedures for introduction of MADM cassettes into new loci in the mouse genome, and describe new approaches for expanding the utility of MADM. We show that: 1) Targeted homologous recombination outperforms random transgenesis in generation of reliably expressed MADM cassettes, 2) MADM cassettes in new genomic loci need to be validated for biallelic and ubiquitous expression, 3) Recombination between MADM cassettes on different chromosomes can be used to study reciprocal chromosomal deletions/duplications, and 4) MADM can be modified to permit transgene expression by combining it with a binary expression system. The advances described in this study expand current, and enable new and more versatile applications of MADM.

    View details for DOI 10.1371/journal.pone.0033332

    View details for Web of Science ID 000303894900024

    View details for PubMedID 22479386

  • Secreted Semaphorins from Degenerating Larval ORN Axons Direct Adult Projection Neuron Dendrite Targeting NEURON Sweeney, L. B., Chou, Y., Wu, Z., Joo, W., Komiyama, T., Potter, C. J., Kolodkin, A. L., Garcia, K. C., Luo, L. 2011; 72 (5): 734-747

    Abstract

    During assembly of the Drosophila olfactory circuit, projection neuron (PN) dendrites prepattern the developing antennal lobe before the arrival of axons from their presynaptic partners, the adult olfactory receptor neurons (ORNs). We previously found that levels of transmembrane Semaphorin-1a, which acts as a receptor, instruct PN dendrite targeting along the dorsolateral-ventromedial axis. Here we show that two secreted semaphorins, Sema-2a and Sema-2b, provide spatial cues for PN dendrite targeting. Sema-2a and Sema-2b proteins are distributed in gradients opposing the Sema-1a protein gradient, and Sema-1a binds to Sema-2a-expressing cells. In Sema-2a and Sema-2b double mutants, PN dendrites that normally target dorsolaterally in the antennal lobe mistarget ventromedially, phenocopying cell-autonomous Sema-1a removal from these PNs. Cell ablation, cell-specific knockdown, and rescue experiments indicate that secreted semaphorins from degenerating larval ORN axons direct dendrite targeting. Thus, a degenerating brain structure instructs the wiring of a developing circuit through the repulsive action of secreted semaphorins.

    View details for DOI 10.1016/j.neuron.2011.09.026

    View details for Web of Science ID 000297971100009

    View details for PubMedID 22153371

  • Site-specific integrase-mediated transgenesis in mice via pronuclear injection PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA Tasic, B., Hippenmeyer, S., Wang, C., Gamboa, M., Zong, H., Chen-Tsai, Y., Luo, L. 2011; 108 (19): 7902-7907

    Abstract

    Microinjection of recombinant DNA into zygotic pronuclei has been widely used for producing transgenic mice. However, with this method, the insertion site, integrity, and copy number of the transgene cannot be controlled. Here, we present an integrase-based approach to produce transgenic mice via pronuclear injection, whereby an intact single-copy transgene can be inserted into predetermined chromosomal loci with high efficiency (up to 40%), and faithfully transmitted through generations. We show that neighboring transgenic elements and bacterial DNA within the transgene cause profound silencing and expression variability of the transgenic marker. Removal of these undesirable elements leads to global high-level marker expression from transgenes driven by a ubiquitous promoter. We also obtained faithful marker expression from a tissue-specific promoter. The technique presented here will greatly facilitate murine transgenesis and precise structure/function dissection of mammalian gene function and regulation in vivo.

    View details for DOI 10.1073/pnas.1019507108

    View details for Web of Science ID 000290439500052

    View details for PubMedID 21464299

  • The chromatin remodeling factor Bap55 functions through the TIP60 complex to regulate olfactory projection neuron dendrite targeting NEURAL DEVELOPMENT Tea, J. S., Luo, L. 2011; 6

    Abstract

    The Drosophila olfactory system exhibits very precise and stereotyped wiring that is specified predominantly by genetic programming. Dendrites of olfactory projection neurons (PNs) pattern the developing antennal lobe before olfactory receptor neuron axon arrival, indicating an intrinsic wiring mechanism for PN dendrites. These wiring decisions are likely determined through a transcriptional program.We find that loss of Brahma associated protein 55 kD (Bap55) results in a highly specific PN mistargeting phenotype. In Bap55 mutants, PNs that normally target to the DL1 glomerulus mistarget to the DA4l glomerulus with 100% penetrance. Loss of Bap55 also causes derepression of a GAL4 whose expression is normally restricted to a small subset of PNs. Bap55 is a member of both the Brahma (BRM) and the Tat interactive protein 60 kD (TIP60) ATP-dependent chromatin remodeling complexes. The Bap55 mutant phenotype is partially recapitulated by Domino and Enhancer of Polycomb mutants, members of the TIP60 complex. However, distinct phenotypes are seen in Brahma and Snf5-related 1 mutants, members of the BRM complex. The Bap55 mutant phenotype can be rescued by postmitotic expression of Bap55, or its human homologs BAF53a and BAF53b.Our results suggest that Bap55 functions through the TIP60 chromatin remodeling complex to regulate dendrite wiring specificity in PNs. The specificity of the mutant phenotypes suggests a position for the TIP60 complex at the top of a regulatory hierarchy that orchestrates dendrite targeting decisions.

    View details for DOI 10.1186/1749-8104-6-5

    View details for Web of Science ID 000290529200002

    View details for PubMedID 21284845

  • Genetic Mosaic Dissection of Lis1 and Ndel1 in Neuronal Migration NEURON Hippenmeyer, S., Youn, Y. H., Moon, H. M., Miyamichi, K., Zong, H., Wynshaw-Boris, A., Luo, L. 2010; 68 (4): 695-709

    Abstract

    Coordinated migration of newly born neurons to their prospective target laminae is a prerequisite for neural circuit assembly in the developing brain. The evolutionarily conserved LIS1/NDEL1 complex is essential for neuronal migration in the mammalian cerebral cortex. The cytoplasmic nature of LIS1 and NDEL1 proteins suggest that they regulate neuronal migration cell autonomously. Here, we extend mosaic analysis with double markers (MADM) to mouse chromosome 11 where Lis1, Ndel1, and 14-3-3? (encoding a LIS1/NDEL1 signaling partner) are located. Analyses of sparse and uniquely labeled mutant cells in mosaic animals reveal distinct cell-autonomous functions for these three genes. Lis1 regulates neuronal migration efficiency in a dose-dependent manner, while Ndel1 is essential for a specific, previously uncharacterized, late step of neuronal migration: entry into the target lamina. Comparisons with previous genetic perturbations of Lis1 and Ndel1 also suggest a surprising degree of cell-nonautonomous function for these proteins in regulating neuronal migration.

    View details for DOI 10.1016/j.neuron.2010.09.027

    View details for Web of Science ID 000285079500009

    View details for PubMedID 21092859

  • Ten years of Nature Reviews Neuroscience: insights from the highly cited NATURE REVIEWS NEUROSCIENCE Luo, L., Rodriguez, E., Jerbi, K., Lachaux, J., Martinerie, J., Corbetta, M., Shulman, G. L., Piomelli, D., Turrigiano, G. G., Nelson, S. B., Joels, M., de Kloet, E. R., Holsboer, F., Amodio, D. M., Frith, C. D., Block, M. L., Zecca, L., Hong, J., Dantzer, R., Kelley, K. W., Craig, A. D. 2010; 11 (10): 718-?

    Abstract

    To celebrate the first 10 years of Nature Reviews Neuroscience, we invited the authors of the most cited article of each year to look back on the state of their field of research at the time of publication and the impact their article has had, and to discuss the questions that might be answered in the next 10 years. This selection of highly cited articles provides interesting snapshots of the progress that has been made in diverse areas of neuroscience. They show the enormous influence of neuroimaging techniques and highlight concepts that have generated substantial interest in the past decade, such as neuroimmunology, social neuroscience and the 'network approach' to brain function. These advancements will pave the way for further exciting discoveries that lie ahead.

    View details for DOI 10.1038/nrn2912

    View details for Web of Science ID 000281928500005

    View details for PubMedID 20852655

  • Patterning Axon Targeting of Olfactory Receptor Neurons by Coupled Hedgehog Signaling at Two Distinct Steps CELL Chou, Y., Zheng, X., Beachy, P. A., Luo, L. 2010; 142 (6): 954-966

    Abstract

    We present evidence for a coupled two-step action of Hedgehog signaling in patterning axon targeting of Drosophila olfactory receptor neurons (ORNs). In the first step, differential Hedgehog pathway activity in peripheral sensory organ precursors creates ORN populations with different levels of the Patched receptor. Different Patched levels in ORNs then determine axonal responsiveness to target-derived Hedgehog in the brain: only ORN axons that do not express high levels of Patched are responsive to and require a second step of Hedgehog signaling for target selection. Hedgehog signaling in the imaginal sensory organ precursors thus confers differential ORN responsiveness to Hedgehog-mediated axon targeting in the brain. This mechanism contributes to the spatial coordination of ORN cell bodies in the periphery and their glomerular targets in the brain. Such coupled two-step signaling may be more generally used to coordinate other spatially and temporally segregated developmental events.

    View details for DOI 10.1016/j.cell.2010.08.015

    View details for Web of Science ID 000281855000017

    View details for PubMedID 20850015

  • 'Fore Brain: A Hint of the Ancestral Cortex CELL Sweeney, L. B., Luo, L. 2010; 142 (5): 679-681

    Abstract

    By combining gene expression profiling with image registration, Tomer et al. (2010) find that the mushroom body of the segmented worm Platynereis dumerilii shares many features with the mammalian cerebral cortex. The authors propose that the mushroom body and cortex evolved from the same structure in the common ancestor of vertebrates and invertebrates.

    View details for DOI 10.1016/j.cell.2010.08.024

    View details for Web of Science ID 000281523200011

    View details for PubMedID 20813256

  • Histone Deacetylase Rpd3 Regulates Olfactory Projection Neuron Dendrite Targeting via the Transcription Factor Prospero JOURNAL OF NEUROSCIENCE Tea, J. S., Chihara, T., Luo, L. 2010; 30 (29): 9939-9946

    Abstract

    Compared to the mechanisms of axon guidance, relatively little is known about the transcriptional control of dendrite guidance. The Drosophila olfactory system with its stereotyped organization provides an excellent model to study the transcriptional control of dendrite wiring specificity. Each projection neuron (PN) targets its dendrites to a specific glomerulus in the antennal lobe and its axon stereotypically to higher brain centers. Using a forward genetic screen, we identified a mutation in Rpd3 that disrupts PN targeting specificity. Rpd3 encodes a class I histone deacetylase (HDAC) homologous to mammalian HDAC1 and HDAC2. Rpd3(-/-) PN dendrites that normally target to a dorsolateral glomerulus mistarget to medial glomeruli in the antennal lobe, and axons exhibit a severe overbranching phenotype. These phenotypes can be rescued by postmitotic expression of Rpd3 but not HDAC3, the only other class I HDAC in Drosophila. Furthermore, disruption of the atypical homeodomain transcription factor Prospero (Pros) yields similar phenotypes, which can be rescued by Pros expression in postmitotic neurons. Strikingly, overexpression of Pros can suppress Rpd3(-/-) phenotypes. Our study suggests a specific function for the general chromatin remodeling factor Rpd3 in regulating dendrite targeting in neurons, largely through the postmitotic action of the Pros transcription factor.

    View details for DOI 10.1523/JNEUROSCI.1643-10.2010

    View details for Web of Science ID 000280206500030

    View details for PubMedID 20660276

  • Visualizing the Distribution of Synapses from Individual Neurons in the Mouse Brain PLOS ONE Li, L., Tasic, B., Micheva, K. D., Ivanov, V. M., Spletter, M. L., Smith, S. J., Luo, L. 2010; 5 (7)

    Abstract

    Proper function of the mammalian brain relies on the establishment of highly specific synaptic connections among billions of neurons. To understand how complex neural circuits function, it is crucial to precisely describe neuronal connectivity and the distributions of synapses to and from individual neurons.In this study, we present a new genetic synaptic labeling method that relies on expression of a presynaptic marker, synaptophysin-GFP (Syp-GFP) in individual neurons in vivo. We assess the reliability of this method and use it to analyze the spatial patterning of synapses in developing and mature cerebellar granule cells (GCs). In immature GCs, Syp-GFP is distributed in both axonal and dendritic regions. Upon maturation, it becomes strongly enriched in axons. In mature GCs, we analyzed synapses along their ascending segments and parallel fibers. We observe no differences in presynaptic distribution between GCs born at different developmental time points and thus having varied depths of projections in the molecular layer. We found that the mean densities of synapses along the parallel fiber and the ascending segment above the Purkinje cell (PC) layer are statistically indistinguishable, and higher than previous estimates. Interestingly, presynaptic terminals were also found in the ascending segments of GCs below and within the PC layer, with the mean densities two-fold lower than that above the PC layer. The difference in the density of synapses in these parts of the ascending segment likely reflects the regional differences in postsynaptic target cells of GCs.The ability to visualize synapses of single neurons in vivo is valuable for studying synaptogenesis and synaptic plasticity within individual neurons as well as information flow in neural circuits.

    View details for DOI 10.1371/journal.pone.0011503

    View details for Web of Science ID 000279715300014

    View details for PubMedID 20634890

  • The Q System: A Repressible Binary System for Transgene Expression, Lineage Tracing, and Mosaic Analysis CELL Potter, C. J., Tasic, B., Russler, E. V., Liang, L., Luo, L. 2010; 141 (3): 536-548

    Abstract

    We describe a new repressible binary expression system based on the regulatory genes from the Neurospora qa gene cluster. This "Q system" offers attractive features for transgene expression in Drosophila and mammalian cells: low basal expression in the absence of the transcriptional activator QF, high QF-induced expression, and QF repression by its repressor QS. Additionally, feeding flies quinic acid can relieve QS repression. The Q system offers many applications, including (1) intersectional "logic gates" with the GAL4 system for manipulating transgene expression patterns, (2) GAL4-independent MARCM analysis, and (3) coupled MARCM analysis to independently visualize and genetically manipulate siblings from any cell division. We demonstrate the utility of the Q system in determining cell division patterns of a neuronal lineage and gene function in cell growth and proliferation, and in dissecting neurons responsible for olfactory attraction. The Q system can be expanded to other uses in Drosophila and to any organism conducive to transgenesis.

    View details for DOI 10.1016/j.cell.2010.02.025

    View details for Web of Science ID 000277180800023

    View details for PubMedID 20434990

  • The olfactory circuit of the fruit fly Drosophila melanogaster SCIENCE CHINA-LIFE SCIENCES Liang, L., Luo Liqun, L. Q. 2010; 53 (4): 472-484

    Abstract

    The olfactory circuit of the fruit fly Drosophila melanogaster has emerged in recent years as an excellent paradigm for studying the principles and mechanisms of information processing in neuronal circuits. We discuss here the organizational principles of the olfactory circuit that make it an attractive model for experimental manipulations, the lessons that have been learned, and future challenges.

    View details for DOI 10.1007/s11427-010-0099-z

    View details for Web of Science ID 000277417800011

  • Diversity and wiring variability of olfactory local interneurons in the Drosophila antennal lobe NATURE NEUROSCIENCE Chou, Y., Spletter, M. L., Yaksi, E., Leong, J. C., Wilson, R. I., Luo, L. 2010; 13 (4): 439-U60

    Abstract

    Local interneurons are essential in information processing by neural circuits. Here we present a comprehensive genetic, anatomical and electrophysiological analysis of local interneurons (LNs) in the Drosophila melanogaster antennal lobe, the first olfactory processing center in the brain. We found LNs to be diverse in their neurotransmitter profiles, connectivity and physiological properties. Analysis of >1,500 individual LNs revealed principal morphological classes characterized by coarsely stereotyped glomerular innervation patterns. Some of these morphological classes showed distinct physiological properties. However, the finer-scale connectivity of an individual LN varied considerably across brains, and there was notable physiological variability within each morphological or genetic class. Finally, LN innervation required interaction with olfactory receptor neurons during development, and some individual variability also likely reflected LN-LN interactions. Our results reveal an unexpected degree of complexity and individual variation in an invertebrate neural circuit, a result that creates challenges for solving the Drosophila connectome.

    View details for DOI 10.1038/nn.2489

    View details for Web of Science ID 000276073500013

    View details for PubMedID 20139975

  • Dendritic tiling through TOR signalling EMBO JOURNAL Hong, W., Luo, L. 2009; 28 (24): 3783-3784

    View details for DOI 10.1038/emboj.2009.353

    View details for Web of Science ID 000272833700001

    View details for PubMedID 20010972

  • Leucine-rich repeat transmembrane proteins instruct discrete dendrite targeting in an olfactory map NATURE NEUROSCIENCE Hong, W., Zhu, H., Potter, C. J., Barsh, G., Kurusu, M., Zinn, K., Luo, L. 2009; 12 (12): 1542-U89

    Abstract

    Olfactory systems utilize discrete neural pathways to process and integrate odorant information. In Drosophila, axons of first-order olfactory receptor neurons (ORNs) and dendrites of second-order projection neurons (PNs) form class-specific synaptic connections at approximately 50 glomeruli. The mechanisms underlying PN dendrite targeting to distinct glomeruli in a three-dimensional discrete neural map are unclear. We found that the leucine-rich repeat (LRR) transmembrane protein Capricious (Caps) was differentially expressed in different classes of PNs. Loss-of-function and gain-of-function studies indicated that Caps instructs the segregation of Caps-positive and Caps-negative PN dendrites to discrete glomerular targets. Moreover, Caps-mediated PN dendrite targeting was independent of presynaptic ORNs and did not involve homophilic interactions. The closely related protein Tartan was partially redundant with Caps. These LRR proteins are probably part of a combinatorial cell-surface code that instructs discrete olfactory map formation.

    View details for DOI 10.1038/nn.2442

    View details for Web of Science ID 000272065600014

    View details for PubMedID 19915565

  • Neuroscience. Brain wiring by presorting axons. Science Miyamichi, K., Luo, L. 2009; 325 (5940): 544-545

    View details for DOI 10.1126/science.1178117

    View details for PubMedID 19644096

  • Uncoupling Dendrite Growth and Patterning: Single-Cell Knockout Analysis of NMDA Receptor 2B NEURON Espinosa, J. S., Wheeler, D. G., Tsien, R. W., Luo, L. 2009; 62 (2): 205-217

    Abstract

    N-methyl-D-aspartate receptors (NMDARs) play important functions in neural development. NR2B is the predominant NR2 subunit of NMDAR in the developing brain. Here we use mosaic analysis with double markers (MADM) to knock out NR2B in isolated single cells and analyze its cell-autonomous function in dendrite development. NR2B mutant dentate gyrus granule cells (dGCs) and barrel cortex layer 4 spiny stellate cells (bSCs) have similar dendritic growth rates, total length, and branch number as control cells. However, mutant dGCs maintain supernumerary primary dendrites resulting from a pruning defect. Furthermore, while control bSCs restrict dendritic growth to a single barrel, mutant bSCs maintain dendritic growth in multiple barrels. Thus, NR2B functions cell autonomously to regulate dendrite patterning to ensure that sensory information is properly represented in the cortex. Our study also indicates that molecular mechanisms that regulate activity-dependent dendrite patterning can be separated from those that control general dendrite growth and branching.

    View details for DOI 10.1016/j.neuron.2009.03.006

    View details for Web of Science ID 000265774100009

    View details for PubMedID 19409266

  • A New Family of Odorant Receptors in Drosophila CELL Spletter, M. L., Luo, L. 2009; 136 (1): 23-25

    Abstract

    In the fruit fly Drosophila, not all olfactory sensory neurons express a seven transmembrane odorant receptor, suggesting that other types of odorant receptors might exist. Benton et al. (2009) now present evidence that a family of proteins related to ionotropic glutamate receptors is a previously unrecognized class of odorant receptors.

    View details for DOI 10.1016/j.cell.2008.12.031

    View details for Web of Science ID 000262318400011

    View details for PubMedID 19135885

  • MicroRNA Processing Pathway Regulates Olfactory Neuron Morphogenesis CURRENT BIOLOGY Berdnik, D., Fan, A. P., Potter, C. J., Luo, L. 2008; 18 (22): 1754-1759

    Abstract

    The microRNA (miRNA) processing pathway produces miRNAs as posttranscriptional regulators of gene expression. The nuclear RNase III Drosha catalyzes the first processing step together with the dsRNA binding protein DGCR8/Pasha generating pre-miRNAs [1, 2]. The next cleavage employs the cytoplasmic RNase III Dicer producing miRNA duplexes [3, 4]. Finally, Argonautes are recruited with miRNAs into an RNA-induced silencing complex for mRNA recognition (Figure 1A). Here, we identify two members of the miRNA pathway, Pasha and Dicer-1, in a forward genetic screen for mutations that disrupt wiring specificity of Drosophila olfactory projection neurons (PNs). The olfactory system is built as discrete map of highly stereotyped neuronal connections [5, 6]. Each PN targets dendrites to a specific glomerulus in the antennal lobe and projects axons stereotypically into higher brain centers [7-9]. In selected PN classes, pasha and Dicer-1 mutants cause specific PN dendrite mistargeting in the antennal lobe and altered axonal terminations in higher brain centers. Furthermore, Pasha and Dicer-1 act cell autonomously in postmitotic neurons to regulate dendrite and axon targeting during development. However, Argonaute-1 and Argonaute-2 are dispensable for PN morphogenesis. Our findings suggest a role for the miRNA processing pathway in establishing wiring specificity in the nervous system.

    View details for DOI 10.1016/j.cub.2008.09.045

    View details for Web of Science ID 000261244800025

    View details for PubMedID 19013069

  • Octopamine fuels fighting flies NATURE NEUROSCIENCE Potter, C. J., Luo, L. 2008; 11 (9): 989-990

    Abstract

    The neural basis of aggression is poorly understood. A study in this issue used genetic scalpels to dissect the circuitry of the fly brain and identified a small cluster of octopaminergic neurons that can make a fly fighting mad.

    View details for DOI 10.1038/nn0908-989

    View details for Web of Science ID 000258720000003

    View details for PubMedID 18725900

  • Genomic analysis of Drosophila neuronal remodeling: A role for the RNA-binding protein boule as a negative regulator of axon pruning JOURNAL OF NEUROSCIENCE Hoopfer, E. D., Penton, A., Watts, R. J., Luo, L. 2008; 28 (24): 6092-6103

    Abstract

    Drosophila mushroom body (MB) gamma neurons undergo axon pruning during metamorphosis through a process of localized degeneration of specific axon branches. Developmental axon degeneration is initiated by the steroid hormone ecdysone, acting through a nuclear receptor complex composed of USP (ultraspiracle) and EcRB1 (ecdysone receptor B1) to regulate gene expression in MB gamma neurons. To identify ecdysone-dependent gene expression changes in MB gamma neurons at the onset of axon pruning, we use laser capture microdissection to isolate wild-type and mutant MB neurons in which EcR (ecdysone receptor) activity is genetically blocked, and analyze expression changes by microarray. We identify several molecular pathways that are regulated in MB neurons by ecdysone. The most striking observation is the upregulation of genes involved in the UPS (ubiquitin-proteasome system), which is cell autonomously required for gamma neuron pruning. In addition, we characterize the function of Boule, an evolutionarily conserved RNA-binding protein previously implicated in spermatogenesis in flies and vertebrates. boule expression is downregulated by ecdysone in MB neurons at the onset of pruning, and forced expression of Boule in MB gamma neurons is sufficient to inhibit axon pruning. This activity is dependent on the RNA-binding domain of Boule and a conserved DAZ (deleted in azoospermia) domain implicated in interactions with other RNA-binding proteins. However, loss of Boule does not result in obvious defects in axon pruning or morphogenesis of MB neurons, suggesting that it acts redundantly with other ecdyonse-regulated genes. We propose a novel function for Boule in the CNS as a negative regulator of developmental axon pruning.

    View details for DOI 10.1523/JNEUROSCI.0677-08.2008

    View details for Web of Science ID 000256668500004

    View details for PubMedID 18550751

  • Genetic dissection of neural circuits NEURON Luo, L., Callaway, E. M., Svoboda, K. 2008; 57 (5): 634-660

    Abstract

    Understanding the principles of information processing in neural circuits requires systematic characterization of the participating cell types and their connections, and the ability to measure and perturb their activity. Genetic approaches promise to bring experimental access to complex neural systems, including genetic stalwarts such as the fly and mouse, but also to nongenetic systems such as primates. Together with anatomical and physiological methods, cell-type-specific expression of protein markers and sensors and transducers will be critical to construct circuit diagrams and to measure the activity of genetically defined neurons. Inactivation and activation of genetically defined cell types will establish causal relationships between activity in specific groups of neurons, circuit function, and animal behavior. Genetic analysis thus promises to reveal the logic of the neural circuits in complex brains that guide behaviors. Here we review progress in the genetic analysis of neural circuits and discuss directions for future research and development.

    View details for DOI 10.1016/j.neuron.2008.01.002

    View details for Web of Science ID 000254077300005

    View details for PubMedID 18341986

  • Timing neurogenesis and differentiation: Insights from quantitative clonal analyses of cerebellar granule cells JOURNAL OF NEUROSCIENCE Espinosa, J. S., Luo, L. 2008; 28 (10): 2301-2312

    Abstract

    The cerebellum is an excellent model system to study how developmental programs give rise to exquisite neuronal circuits in the adult brain. Here, we describe our findings regarding granule cell neurogenesis and differentiation using the MADM method (mosaic analysis with double markers) in mice. By following the development of individual granule cell clones, we show that (1) granule cell precursors (GCPs) undergo predominantly symmetric division during postnatal development; (2) clonally related granule cells (GCs) exit the cell cycle within a narrow time window and stack their axons in the molecular layer in chronological order from deep to superficial sublayers; and (3) whereas the average GCP proliferation in the external granular layer is progressively slower as development proceeds, there is a rapid expansion of GCPs shortly before clonally related GCs exit the cell cycle. These properties produce GC clones that are distinct, each having a restricted axonal projection, but that are on average similar in cell number. We discuss possible developmental mechanisms and functional implications of these findings.

    View details for DOI 10.1523/JNEUROSCI.5157-07.2008

    View details for Web of Science ID 000253818800002

    View details for PubMedID 18322077

  • piggyBac-based mosaic screen identifies a postmitotic function for cohesin in regulating developmental axon pruning DEVELOPMENTAL CELL Schuldiner, O., Berdnik, D., Levy, J. M., Wu, J. S., Luginbuhl, D., Camille Gontang, A., Luo, L. 2008; 14 (2): 227-238

    Abstract

    Developmental axon pruning is widely used to refine neural circuits. We performed a mosaic screen to identify mutations affecting axon pruning of Drosophila mushroom body gamma neurons. We constructed a modified piggyBac vector with improved mutagenicity and generated insertions in >2000 genes. We identified two cohesin subunits (SMC1 and SA) as being essential for axon pruning. The cohesin complex maintains sister-chromatid cohesion during cell division in eukaryotes. However, we show that the pruning phenotype in SMC1(-/-) clones is rescued by expressing SMC1 in neurons, revealing a postmitotic function. SMC1(-/-) clones exhibit reduced levels of the ecdysone receptor EcR-B1, a key regulator of axon pruning. The pruning phenotype is significantly suppressed by overexpressing EcR-B1 and is enhanced by a reduced dose of EcR, supporting a causal relationship. We also demonstrate a postmitotic role for SMC1 in dendrite targeting of olfactory projection neurons. We suggest that cohesin regulates diverse aspects of neuronal morphogenesis.

    View details for DOI 10.1016/j.devce1.2007.11.001

    View details for Web of Science ID 000253241400012

    View details for PubMedID 18267091

  • Development of continuous and discrete neural maps NEURON Luo, L., Flanagan, J. G. 2007; 56 (2): 284-300

    Abstract

    Two qualitatively different kinds of neural map have been described: continuous maps exemplified by the visual retinotopic map, and discrete maps exemplified by the olfactory glomerular map. Here, we review developmental mechanisms of retinotopic and olfactory glomerular mapping and discuss underlying commonalities that have emerged from recent studies. These include the use of molecular gradients, axon-axon interactions, and the interplay between labeling molecules and neuronal activity in establishing these maps. Since visual retinotopic and olfactory glomerular maps represent two ends of a continuum that includes many other types of neural map in between, these emerging general principles may be widely applicable to map formation throughout the nervous system.

    View details for DOI 10.1016/j.neuron.2007.10.014

    View details for Web of Science ID 000250740700007

    View details for PubMedID 17964246

  • Fly MARCM and mouse MADM: Genetic methods of labeling and manipulating single neurons BRAIN RESEARCH REVIEWS Luo, L. 2007; 55 (2): 220-227

    Abstract

    The Golgi staining method has served neuroscience well for more than a century. In this assay I review recent progresses using genetic methods to recapitulate and extend the Golgi staining method. These methods enable new discoveries on organization and development of neuronal circuits in the fly and mouse brains.

    View details for Web of Science ID 000250952300004

    View details for PubMedID 17408568

  • A global double-fluorescent cre reporter mouse GENESIS Muzumdar, M. D., Tasic, B., Miyamichi, K., Li, L., Luo, L. 2007; 45 (9): 593-605

    Abstract

    The Cre/loxP system has been used extensively for conditional mutagenesis in mice. Reporters of Cre activity are important for defining the spatial and temporal extent of Cre-mediated recombination. Here we describe mT/mG, a double-fluorescent Cre reporter mouse that expresses membrane-targeted tandem dimer Tomato (mT) prior to Cre-mediated excision and membrane-targeted green fluorescent protein (mG) after excision. We show that reporter expression is nearly ubiquitous, allowing visualization of fluorescent markers in live and fixed samples of all tissues examined. We further demonstrate that mG labeling is Cre-dependent, complementary to mT at single cell resolution, and distinguishable by fluorescence-activated cell sorting. Both membrane-targeted markers outline cell morphology, highlight membrane structures, and permit visualization of fine cellular processes. In addition to serving as a global Cre reporter, the mT/mG mouse may also be used as a tool for lineage tracing, transplantation studies, and analysis of cell morphology in vivo.

    View details for DOI 10.1002/dvg.20335

    View details for Web of Science ID 000250365600008

    View details for PubMedID 17868096

  • Lola regulates Drosophila olfactory projection neuron identity and targeting specificity NEURAL DEVELOPMENT Spletter, M. L., Liu, J., Liu, J., Su, H., Giniger, E., Komiyama, T., Quake, S., Luo, L. 2007; 2

    Abstract

    Precise connections of neural circuits can be specified by genetic programming. In the Drosophila olfactory system, projection neurons (PNs) send dendrites to single glomeruli in the antenna lobe (AL) based upon lineage and birth order and send axons with stereotyped terminations to higher olfactory centers. These decisions are likely specified by a PN-intrinsic transcriptional code that regulates the expression of cell-surface molecules to instruct wiring specificity.We find that the loss of longitudinals lacking (lola), which encodes a BTB-Zn-finger transcription factor with 20 predicted splice isoforms, results in wiring defects in both axons and dendrites of all lineages of PNs. RNA in situ hybridization and quantitative RT-PCR suggest that most if not all lola isoforms are expressed in all PNs, but different isoforms are expressed at widely varying levels. Overexpression of individual lola isoforms fails to rescue the lola null phenotypes and causes additional phenotypes. Loss of lola also results in ectopic expression of Gal4 drivers in multiple cell types and in the loss of transcription factor gene lim1 expression in ventral PNs.Our results indicate that lola is required for wiring of axons and dendrites of most PN classes, and suggest a need for its molecular diversity. Expression pattern changes of Gal4 drivers in lola-/- clones imply that lola normally represses the expression of these regulatory elements in a subset of the cells surrounding the AL. We propose that Lola functions as a general transcription factor that regulates the expression of multiple genes ultimately controlling PN identity and wiring specificity.

    View details for DOI 10.1186/1749-8104-2-14

    View details for Web of Science ID 000258981200001

    View details for PubMedID 17634136

  • Cytoplasmic and mitochondrial protein translation in axonal and dendritic terminal arborization NATURE NEUROSCIENCE Chihara, T., Luginbuhl, D., Luo, L. 2007; 10 (7): 828-837

    Abstract

    We identified a mutation in Aats-gly (also known as gars or glycyl-tRNA synthetase), the Drosophila melanogaster ortholog of the human GARS gene that is associated with Charcot-Marie-Tooth neuropathy type 2D (CMT2D), from a mosaic genetic screen. Loss of gars in Drosophila neurons preferentially affects the elaboration and stability of terminal arborization of axons and dendrites. The human and Drosophila genes each encode both a cytoplasmic and a mitochondrial isoform. Using additional mutants that selectively disrupt cytoplasmic or mitochondrial protein translation, we found that cytoplasmic protein translation is required for terminal arborization of both dendrites and axons during development. In contrast, disruption of mitochondrial protein translation preferentially affects the maintenance of dendritic arborization in adults. We also provide evidence that human GARS shows equivalent functions in Drosophila, and that CMT2D causal mutations show loss-of-function properties. Our study highlights different demands of protein translation for the development and maintenance of axons and dendrites.

    View details for DOI 10.1038/nn1910

    View details for Web of Science ID 000247560200012

    View details for PubMedID 17529987

  • Comprehensive maps of Drosophila higher offactory centers: Spatially segregated fruit and pheromone representation CELL Jefferis, G. S., Potter, C. J., Chan, A. I., Marin, E. C., Rohlfing, T., Maurer, C. R., Luo, L. 2007; 128 (6): 1187-1203

    Abstract

    In Drosophila, approximately 50 classes of olfactory receptor neurons (ORNs) send axons to 50 corresponding glomeruli in the antennal lobe. Uniglomerular projection neurons (PNs) relay olfactory information to the mushroom body (MB) and lateral horn (LH). Here, we combine single-cell labeling and image registration to create high-resolution, quantitative maps of the MB and LH for 35 input PN channels and several groups of LH neurons. We find (1) PN inputs to the MB are stereotyped as previously shown for the LH; (2) PN partners of ORNs from different sensillar groups are clustered in the LH; (3) fruit odors are represented mostly in the posterior-dorsal LH, whereas candidate pheromone-responsive PNs project to the anterior-ventral LH; (4) dendrites of single LH neurons each overlap with specific subsets of PN axons. Our results suggest that the LH is organized according to biological values of olfactory input.

    View details for DOI 10.1016/j.cell.2007.01.040

    View details for Web of Science ID 000245396200023

    View details for PubMedID 17382886

  • Modeling sporadic loss of heterozygosity in mice by using mosaic analysis with double markers (MADM) PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA Muzumdar, M. D., Luo, L., Zong, H. 2007; 104 (11): 4495-4500

    Abstract

    The initiation and progression of many human cancers involve either somatic activation of protooncogenes or inactivation of tumor-suppressor genes (TSGs) in sporadic cells. Although sporadic gain-of-function of protooncogenes has been successfully modeled in mice [e.g., Johnson L, Mercer K, Greenbaum D, Bronson RT, Crowley D, Tuveson DA, Jacks T (2001) Nature 410:1111-1116], generating a similar degree of sparseness of TSG loss-of-function remains a challenge. Here, we use mosaic analysis with double markers (MADM) to achieve TSG inactivation and concurrent labeling in sporadic somatic cells of mice, closely mimicking loss of heterozygosity as occurs in human cancers. As proof of principle, we studied the consequence of sporadic loss of p27kip1, a cyclin-dependent kinase inhibitor. MADM-mediated loss of p27kip1 results in mutant cell expansion markedly greater than that observed in conventional p27kip1 knockouts. Moreover, the direct comparison of WT and mutant cells at single-cell resolution afforded by MADM reveals that p27kip1 regulates organ size in vivo by cell-autonomous control of cell cycle exit timing. These studies establish MADM as a high-resolution method for modeling sporadic loss of heterozygosity in mice, providing insights into TSG function.

    View details for DOI 10.1073/pnas.0606491104

    View details for Web of Science ID 000244972700046

    View details for PubMedID 17360552

  • Intrinsic control of precise dendritic targeting by an ensemble of transcription factors CURRENT BIOLOGY Komiyama, T., Luo, L. 2007; 17 (3): 278-285

    Abstract

    Proper information processing in neural circuits requires establishment of specific connections between pre- and postsynaptic neurons. Targeting specificity of neurons is instructed by cell-surface receptors on the growth cones of axons and dendrites, which confer responses to external guidance cues. Expression of cell-surface receptors is in turn regulated by neuron-intrinsic transcriptional programs. In the Drosophila olfactory system, each projection neuron (PN) achieves precise dendritic targeting to one of 50 glomeruli in the antennal lobe. PN dendritic targeting is specified by lineage and birth order , and their initial targeting occurs prior to contact with axons of their presynaptic partners, olfactory receptor neurons. We search for transcription factors (TFs) that control PN-intrinsic mechanisms of dendritic targeting. We previously identified two POU-domain TFs, acj6 and drifter, as essential players. After testing 13 additional candidates, we identified four TFs (LIM-homeodomain TFs islet and lim1, the homeodomain TF cut, and the zinc-finger TF squeeze) and the LIM cofactor Chip that are required for PN dendritic targeting. These results begin to provide insights into the global strategy of how an ensemble of TFs regulates wiring specificity of a large number of neurons constituting a neural circuit.

    View details for Web of Science ID 000244164700029

    View details for PubMedID 17276922

  • Graded expression of Semaphorin-1a cell-autonomously directs dendritic targeting of olfactory projection neurons CELL Komiyama, T., Sweeney, L. B., Schuldiner, O., Garcia, K. C., Luo, L. 2007; 128 (2): 399-410

    Abstract

    Gradients of axon guidance molecules instruct the formation of continuous neural maps, such as the retinotopic map in the vertebrate visual system. Here we show that molecular gradients can also instruct the formation of a discrete neural map. In the fly olfactory system, axons of 50 classes of olfactory receptor neurons (ORNs) and dendrites of 50 classes of projection neurons (PNs) form one-to-one connections at discrete units called glomeruli. We provide expression, loss- and gain-of-function data to demonstrate that the levels of transmembrane Semaphorin-1a (Sema-1a), acting cell-autonomously as a receptor or part of a receptor complex, direct the dendritic targeting of PNs along the dorsolateral to ventromedial axis of the antennal lobe. Sema-1a also regulates PN axon targeting in higher olfactory centers. Thus, graded expression of Sema-1a contributes to connection specificity from ORNs to PNs and then to higher brain centers, ensuring proper representation of olfactory information in the brain.

    View details for DOI 10.1016/j.cell.2006.12.028

    View details for Web of Science ID 000244420500022

    View details for PubMedID 17254975

  • Temporal target restriction of olfactory receptor neurons by Semaphorin-1a/PlexinA-mediated axon-axon interactions NEURON Sweeney, L. B., Couto, A., Chou, Y., Berdnik, D., Dickson, B. J., Luo, L., Komiyama, T. 2007; 53 (2): 185-200

    Abstract

    Axon-axon interactions have been implicated in neural circuit assembly, but the underlying mechanisms are poorly understood. Here, we show that in the Drosophila antennal lobe, early-arriving axons of olfactory receptor neurons (ORNs) from the antenna are required for the proper targeting of late-arriving ORN axons from the maxillary palp (MP). Semaphorin-1a is required for targeting of all MP but only half of the antennal ORN classes examined. Sema-1a acts nonautonomously to control ORN axon-axon interactions, in contrast to its cell-autonomous function in olfactory projection neurons. Phenotypic and genetic interaction analyses implicate PlexinA as the Sema-1a receptor in ORN targeting. Sema-1a on antennal ORN axons is required for correct targeting of MP axons within the antennal lobe, while interactions amongst MP axons facilitate their entry into the antennal lobe. We propose that Sema-1a/PlexinA-mediated repulsion provides a mechanism by which early-arriving ORN axons constrain the target choices of late-arriving axons.

    View details for DOI 10.1016/j.neuron.2006.12.022

    View details for Web of Science ID 000245126600006

    View details for PubMedID 17224402

  • Wld(S) protection distinguishes axon degeneration following injury from naturally occurring developmental pruning NEURON Hoopfer, E. D., McLaughlin, T., Watts, R. J., Schuldiner, O., O'Leary, D. D., Luo, L. 2006; 50 (6): 883-895

    Abstract

    Axon pruning by degeneration remodels exuberant axonal connections and is widely required for the development of proper circuitry in the nervous system from insects to mammals. Developmental axon degeneration morphologically resembles injury-induced Wallerian degeneration, suggesting similar underlying mechanisms. As previously reported for mice, we show that Wlds protein substantially delays Wallerian degeneration in flies. Surprisingly, Wlds has no effect on naturally occurring developmental axon degeneration in flies or mice, although it protects against injury-induced degeneration of the same axons at the same developmental age. By contrast, the ubiquitin-proteasome system is intrinsically required for both developmental and injury-induced axon degeneration. We also show that the glial cell surface receptor Draper is required for efficient clearance of axon fragments during developmental axon degeneration, similar to its function in injury-induced degeneration. Thus, mechanistically, naturally occurring developmental axon pruning by degeneration and injury-induced axon degeneration differ significantly in early steps, but may converge onto a common execution pathway.

    View details for DOI 10.1016/j.neuron.2006.05.013

    View details for Web of Science ID 000238589500009

    View details for PubMedID 16772170

  • Wiring stability of the adult Drosophila olfactory circuit after lesion JOURNAL OF NEUROSCIENCE Berdnik, D., Chihara, T., Couto, A., Luo, L. Q. 2006; 26 (13): 3367-3376

    Abstract

    Neuronal wiring plasticity in response to experience or injury has been reported in many parts of the adult nervous system. For instance, visual or somatosensory cortical maps can reorganize significantly in response to peripheral lesions, yet a certain degree of stability is essential for neuronal circuits to perform their dedicated functions. Previous studies on lesion-induced neuronal reorganization have primarily focused on systems that use continuous neural maps. Here, we assess wiring plasticity in a discrete neural map represented by the adult Drosophila olfactory circuit. Using conditional expression of toxins, we genetically ablated specific classes of neurons and examined the consequences on their synaptic partners or neighboring classes in the adult antennal lobe. We find no alteration of connection specificity between olfactory receptor neurons (ORNs) and their postsynaptic targets, the projection neurons (PNs). Ablating an ORN class maintains PN dendrites within their glomerular borders, and ORN axons normally innervating an adjacent target do not expand. Likewise, ablating PN classes does not alter their partner ORN axon connectivity. Interestingly, an increase in the contralateral ORN axon terminal density occurs in response to the removal of competing ipsilateral ORNs. Therefore, plasticity in this circuit can occur but is confined within a glomerulus, thereby retaining the wiring specificity of ORNs and PNs. We conclude that, although adult olfactory neurons can undergo plastic changes in response to the loss of competition, the olfactory circuit overall is extremely stable in preserving segregated information channels in this discrete map.

    View details for DOI 10.1523/JNEUROSCI.4941-05.2006

    View details for Web of Science ID 000236363400001

    View details for PubMedID 16571743

  • Dendritic patterning by Dscam and synaptic partner matching in the Drosophila antennal lobe NATURE NEUROSCIENCE Zhu, H. T., Hummel, T., Clemens, J. C., Berdnik, D., Zipursky, S. L., Luo, L. Q. 2006; 9 (3): 349-355

    Abstract

    In the olfactory system of Drosophila melanogaster, axons of olfactory receptor neurons (ORNs) and dendrites of second-order projection neurons typically target 1 of approximately 50 glomeruli. Dscam, an immunoglobulin superfamily protein, acts in ORNs to regulate axon targeting. Here we show that Dscam acts in projection neurons and local interneurons to control the elaboration of dendritic fields. The removal of Dscam selectively from projection neurons or local interneurons led to clumped dendrites and marked reduction in their dendritic field size. Overexpression of Dscam in projection neurons caused dendrites to be more diffuse during development and shifted their relative position in adulthood. Notably, the positional shift of projection neuron dendrites caused a corresponding shift of its partner ORN axons, thus maintaining the connection specificity. This observation provides evidence for a pre- and postsynaptic matching mechanism independent of precise glomerular positioning.

    View details for Web of Science ID 000235645600014

    View details for PubMedID 16474389

  • Development of wiring specificity in the olfactory system CURRENT OPINION IN NEUROBIOLOGY Komiyama, T., Luo, L. Q. 2006; 16 (1): 67-73

    Abstract

    The olfactory system discriminates a large number of odorants using precisely wired neural circuits. It offers an excellent opportunity to study mechanisms of neuronal wiring specificity at the single synapse level. Each olfactory receptor neuron typically expresses only one olfactory receptor from many receptor genes (1000 in mice). In mice, this striking singularity appears to be ensured by a negative feedback mechanism. Olfactory receptor neurons expressing the same receptor converge their axons to stereotypical positions with high precision, a feature that is conserved from insects to mammals. Several molecules have recently been identified that control this process, including olfactory receptors themselves in mice. The second order neurons, mitral cells in mammals and projection neurons in insects, have a similar degree of wiring specificity: studies in Drosophila suggest that projection neuron-intrinsic mechanisms regulate their precise dendritic targeting. Finally, recent studies have revealed interactions of different cell types during circuit assembly, including axon-axon interactions among olfactory receptor neurons and dendro-dendritic interactions of projection neurons, that are essential in establishing wiring specificity of the olfactory circuit.

    View details for DOI 10.1016/j.conb.2005.12.002

    View details for Web of Science ID 000236136200010

    View details for PubMedID 16377177

  • A protocol for dissecting Drosophila melanogaster brains for live imaging or immunostaining NATURE PROTOCOLS Wu, J. S., Luo, L. 2006; 1 (4): 2110-2115

    Abstract

    This protocol describes a basic method for dissection and immunofluorescence staining of the Drosophila brain at various developmental stages. The Drosophila brain has become increasingly useful for studies of neuronal wiring and morphogenesis in combination with techniques such as the 'mosaic analysis with a repressible cell marker' (MARCM) system, where single neurons can be followed in live and fixed tissues for high-resolution analysis of wild-type or genetically manipulated cells. Such high-resolution anatomical study of the brain is also important in characterizing the organization of neural circuits using genetic tools such as GAL4 enhancer trap lines, as Drosophila has been intensively used for studying the neural basis of behavior. Advantages of fluorescence immunostaining include compatibility with multicolor labeling and confocal or multiphoton imaging. This brain dissection and immunofluorescence staining protocol requires approximately 2 to 6 d to complete.

    View details for DOI 10.1038/nprot.2006.336

    View details for Web of Science ID 000251155500055

    View details for PubMedID 17487202

  • A protocol for mosaic analysis with a repressible cell marker (MARCM) in Drosophila NATURE PROTOCOLS Wu, J. S., Luo, L. 2006; 1 (6): 2583-2589

    Abstract

    Mosaic analysis with a repressible cell marker (MARCM) is a genetic technique used in Drosophila to label single cells or multiple cells sharing a single progenitor. Labeled homozygous mutant cells can be generated in an otherwise unlabeled heterozygous animal. Mutant or wild-type labeled cells can also be made to express one or more transgenes. Major applications of MARCM include (i) lineage analysis, (ii) investigating gene function in single or small populations of cells and (iii) neuronal circuit tracing. Our laboratory uses MARCM primarily to label and genetically manipulate neurons; however, this protocol can be adapted to any cell of interest. The protocol involves generating two fly stocks with the necessary genetic elements for MARCM analysis and subsequently generating MARCM clones. Labeled clones can be followed in live and fixed tissues for high-resolution analysis of wild-type or genetically manipulated cells.NOTE: In the PDF version of this article initially published online, the first "FRT" and the "Mutation" labels in Figure 1b were transposed. In both the PDF and HTML versions, "mutant" was omitted from the label on the right, which should read "Labeled homozygous mutant daughter cell". The figure has been corrected in all versions of the article.

    View details for DOI 10.1038/nprot.2006.320

    View details for Web of Science ID 000251155700008

    View details for PubMedID 17406512

  • Glomerular maps without cellular redundancy at successive levels of the Drosophila larval olfactory circuit CURRENT BIOLOGY Ramaekers, A., Magnenat, E., Marin, E. C., Gendre, N., Jefferis, G. S., Luo, L. Q., Stocker, R. F. 2005; 15 (11): 982-992

    Abstract

    Drosophila larvae possess only 21 odorant-receptor neurons (ORNs), whereas adults have 1,300. Does this suggest that the larval olfactory system is built according to a different design than its adult counterpart, or is it just a miniature version thereof?By genetically labeling single neurons with FLP-out and MARCM techniques, we analyze the connectivity of the larval olfactory circuit. We show that each of the 21 ORNs is unique and projects to one of 21 morphologically identifiable antennal-lobe glomeruli. Each glomerulus seems to be innervated by a single projection neuron. Each projection neuron sends its axon to one or two of about 28 glomeruli in the mushroom-body calyx. We have discovered at least seven types of projection neurons that stereotypically link an identified antennal-lobe glomerulus with an identified calycal glomerulus and thus create an olfactory map in a higher brain center.The basic design of the larval olfactory system is similar to the adult one. However, ORNs and projection neurons lack cellular redundancy and do not exhibit any convergent or divergent connectivity; 21 ORNs confront essentially similar numbers of antennal-lobe glomeruli, projection neurons, and calycal glomeruli. Hence, we propose the Drosophila larva as an "elementary" olfactory model system.

    View details for DOI 10.1016/j.cub.2005.04.032

    View details for Web of Science ID 000229984100019

    View details for PubMedID 15936268

  • Mosaic analysis with double markers in mice CELL Zong, H., Espinosa, S., Su, H. H., Muzumdar, M. D., Luo, L. Q. 2005; 121 (3): 479-492

    Abstract

    We describe a method termed MADM (mosaic analysis with double markers) in mice that allows simultaneous labeling and gene knockout in clones of somatic cells or isolated single cells in vivo. Two reciprocally chimeric genes, each containing the N terminus of one marker and the C terminus of the other marker interrupted by a loxP-containing intron, are knocked in at identical locations on homologous chromosomes. Functional expression of markers requires Cre-mediated interchromosomal recombination. MADM reveals that interchromosomal recombination can be induced efficiently in vivo in both mitotic and postmitotic cells in all tissues examined. It can be used to create conditional knockouts in small populations of labeled cells, to determine cell lineage, and to trace neuronal connections. To illustrate the utility of MADM, we show that cerebellar granule cell progenitors are fated at an early stage to produce granule cells with axonal projections limited to specific sublayers of the cerebellar cortex.

    View details for DOI 10.1016/j.cell.2005.02.012

    View details for Web of Science ID 000229031600016

    View details for PubMedID 15882628

  • Function and regulation of Tumbleweed (RacGAP50C) in neuroblast proliferation and neuronal morphogenesis PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA Goldstein, A. Y., Jan, Y. N., Luo, L. Q. 2005; 102 (10): 3834-3839

    Abstract

    Drosophila RacGAP50C and its homologues act as part of a complex with a kinesin-like protein (Pavarotti/Zen-4) that is essential for the formation of the central spindle and completion of cytokinesis [Mishima, M., Kaitna, S. & Glotzer, M. (2002) Dev. Cell 2, 41-54; Somers, W. G. & Saint, R. (2003) Dev. Cell 4, 29-39; Jantsch-Plunger et al. (2000) J. Cell Biol. 149, 1391-1404]. We report here that RacGAP50C corresponds to the tumbleweed (tum) gene previously identified based on its defects in dendrite development of sensory neurons [Gao, F. B., Brenman, J. E., Jan, L. Y. & Jan, Y. N. (1999) Genes Dev. 13, 2549-2561]. Using mushroom body neurogenesis and morphogenesis as a model, we show that Tumbleweed (Tum), Pavarotti, and their association are required for neuroblast proliferation. Tum with a mutation predicted to disrupt the GTPase-activating protein (GAP) activity still largely retains its activity in regulating cell division but is impaired in its activity to limit axon growth. We also provide evidence that Tum and Pavarotti regulate the subcellular localization of each other in postmitotic neurons and that cytoplasmic accumulation of both proteins disrupts axon development in a GAP-dependent manner. Taken together with previous studies of RacGAP50C in regulating cytokinesis, we propose that Tum serves as a scaffolding protein in regulating cell division but acts as a GAP to limit axon growth in postmitotic neurons.

    View details for DOI 10.1073/pnas.0500748102

    View details for Web of Science ID 000227533100057

    View details for PubMedID 15738386

  • Developmentally programmed remodeling of the Drosophila olfactory circuit DEVELOPMENT Marin, E. C., Watts, R. J., Tanaka, N. K., Ito, K., Luo, L. Q. 2005; 132 (4): 725-737

    Abstract

    Neural circuits are often remodeled after initial connections are established. The mechanisms by which remodeling occurs, in particular whether and how synaptically connected neurons coordinate their reorganization, are poorly understood. In Drosophila, olfactory projection neurons (PNs) receive input by synapsing with olfactory receptor neurons in the antennal lobe and relay information to the mushroom body (MB) calyx and lateral horn. Here we show that embryonic-born PNs participate in both the larval and adult olfactory circuits. In the larva, these neurons generally innervate a single glomerulus in the antennal lobe and one or two glomerulus-like substructures in the MB calyx. They persist in the adult olfactory circuit and are prespecified by birth order to innervate a subset of glomeruli distinct from larval-born PNs. Developmental studies indicate that these neurons undergo stereotyped pruning of their dendrites and axon terminal branches locally during early metamorphosis. Electron microscopy analysis reveals that these PNs synapse with MB gamma neurons in the larval calyx and that these synaptic profiles are engulfed by glia during early metamorphosis. As with MB gamma neurons, PN pruning requires cell-autonomous reception of the nuclear hormone ecdysone. Thus, these synaptic partners are independently programmed to prune their dendrites and axons.

    View details for DOI 10.1242/dev.01614

    View details for Web of Science ID 000227427100010

    View details for PubMedID 15659487

  • Development of wiring specificity of the Drosophila olfactory system CHEMICAL SENSES Jefferis, G. S., Marin, E. C., Komiyama, T., Zhu, H. T., Chihara, T., Berdnik, D., Luo, L. Q. 2005; 30: I94-I94

    View details for DOI 10.1093/chemse/bjh130

    View details for Web of Science ID 000227615800047

    View details for PubMedID 15738213

  • Axon retraction and degeneration in development and disease ANNUAL REVIEW OF NEUROSCIENCE Luo, L. Q., O'Leary, D. D. 2005; 28: 127-156

    Abstract

    The selective elimination of axons, dendrites, axon and dendrite branches, and synapses, without loss of the parent neurons, occurs during normal development of the nervous system as well as in response to injury or disease in the adult. The widespread developmental phenomena of exuberant axonal projections and synaptic connections require both small-scale and large-scale axon pruning to generate precise adult connectivity, and they provide a mechanism for neural plasticity in the developing and adult nervous system, as well as a mechanism to evolve differences between species in a projection system. Such pruning is also required to remove axonal connections damaged in the adult, to stabilize the affected neural circuits, and to initiate their repair. Pruning occurs through either retraction or degeneration. Here we review examples of these phenomena and consider potential cellular and molecular mechanisms that underlie axon retraction and degeneration and how they might relate to each other in development and disease.

    View details for DOI 10.1146/annurev.neuro.28.061604.135632

    View details for Web of Science ID 000231235700006

    View details for PubMedID 16022592

  • Rho GTPases regulate axon growth through convergent and divergent signaling pathways NEURON Ng, J. L., Luo, L. Q. 2004; 44 (5): 779-793

    Abstract

    Rho GTPases are essential regulators of cytoskeletal reorganization, but how they do so during neuronal morphogenesis in vivo is poorly understood. Here we show that the actin depolymerization factor cofilin is essential for axon growth in Drosophila neurons. Cofilin function in axon growth is inhibited by LIM kinase and activated by Slingshot phosphatase. Dephosphorylating cofilin appears to be the major function of Slingshot in regulating axon growth in vivo. Genetic data provide evidence that Rho or Rac/Cdc42, via effector kinases Rok or Pak, respectively, activate LIM kinase to inhibit axon growth. Importantly, Rac also activates a Pak-independent pathway that promotes axon growth, and different RacGEFs regulate these distinct pathways. These genetic analyses reveal convergent and divergent pathways from Rho GTPases to the cytoskeleton during axon growth in vivo and suggest that different developmental outcomes could be achieved by biases in pathway selection.

    View details for Web of Science ID 000225548400007

    View details for PubMedID 15572110

  • Like poles repel: Molecular mechanisms of dendritic tiling CELL Chihara, T., Luo, L. Q. 2004; 119 (2): 148-149

    Abstract

    To cover the entire sensory field once and only once, dendrites of some sensory system neurons avoid crossing other dendrites from the same type of neurons. In this issue of Cell, provide first insight into the molecular mechanisms of dendritic tiling.

    View details for Web of Science ID 000224577200002

    View details for PubMedID 15479631

  • Olfactory receptor neuron axon targeting: intrinsic transcriptional control and hierarchical interactions NATURE NEUROSCIENCE Komiyama, T., Carlson, J. R., Luo, L. Q. 2004; 7 (8): 819-825

    Abstract

    From insects to mammals, olfactory receptor neurons (ORNs) expressing a common olfactory receptor target their axons to specific glomeruli with high precision. Here we show in Drosophila that the POU transcription factor Acj6 controls the axon targeting specificity of a subset of ORN classes, as defined by the olfactory receptors that they express. Of these classes, some require Acj6 cell-autonomously, whereas others require Acj6 cell-nonautonomously. Mosaic analyses show that cooperative targeting occurs between axon terminals of the same ORN classes and that there are hierarchical interactions among different ORN classes. We propose that the precision of ORN axon targeting derives from both intrinsic transcriptional control and extensive axon-axon interactions.

    View details for DOI 10.1038/nn1284

    View details for Web of Science ID 000222930800011

    View details for PubMedID 15247920

  • Glia engulf degenerating axons during developmental axon pruning CURRENT BIOLOGY Watts, R. J., Schuldiner, O., Perrino, J., Larsen, C., Luo, L. Q. 2004; 14 (8): 678-684

    Abstract

    Developmental axon pruning is widely used in constructing the nervous system. Accordingly, diverse mechanisms are likely employed for various forms of axon pruning. In the Drosophila mushroom bodies (MB), gamma neurons initially extend axon branches into both the dorsal and medial MB axon lobes in larvae. Through a well-orchestrated set of developmental events during metamorphosis, axon branches to both lobes degenerate prior to the formation of adult connections. Here, we analyze ultrastructural changes underlying axon pruning by using a genetically encoded electron microscopic (EM) marker to selectively label gamma neurons. By inhibiting axon pruning in combination with the use of this EM marker, we demonstrate a causal link between observed cellular events and axon pruning. These events include changes in axon ultrastructure, synaptic degeneration, and engulfment of degenerating axon fragments by glia for their subsequent breakdown via the endosomal-lysosomal pathway. Interestingly, glia selectively invade MB axon lobes at the onset of metamorphosis; this increase in cell number is independent of axon fragmentation. Our study reveals a key role for glia in the removal of axon fragments during developmental axon pruning.

    View details for DOI 10.1016/j.cub.2004.03.035

    View details for Web of Science ID 000220989700019

    View details for PubMedID 15084282

  • Diverse functions of N-cadherin in dendritic and axonal terminal arborization of olfactory projection neurons NEURON Zhu, H. T., Luo, L. Q. 2004; 42 (1): 63-75

    Abstract

    The cadherin superfamily of cell adhesion molecules have been proposed to play important roles in determining synaptic specificity in developing nervous systems. We examine the function of N-cadherin in Drosophila second order olfactory projection neurons (PNs), each of which must selectively target their dendrites to one of approximately 50 glomeruli. Our results do not support an instructive role for N-cadherin in selecting dendritic targets; rather, N-cadherin is essential for PNs to restrict their dendrites to single glomeruli. Mosaic analyses suggest that N-cadherin mediates dendro-dendritic interactions between PNs and thus contributes to refinement of PN dendrites to single glomeruli. N-cadherin is also essential for the development of PN axon terminal arbors in two distinct central targets: regulating branch stability in the lateral horn and restricting high-order branching in the mushroom body. Although the N-cadherin locus potentially encodes eight alternatively spliced isoforms, transgenic expression of one isoform is sufficient to rescue all phenotypes.

    View details for Web of Science ID 000221458400008

    View details for PubMedID 15066265

  • Neuroscience. Calcium and CREST for healthy dendrites. Science Jefferis, G. S., Komiyama, T., Luo, L. 2004; 303 (5655): 179-181

    View details for PubMedID 14715999

  • Developmental origin of wiring specificity in the olfactory system of Drosophila DEVELOPMENT Jefferis, G. S., Vyas, R. M., Berdnik, D., Ramaekers, A., Stocker, R. F., Tanaka, N. K., Ito, K., Luo, L. Q. 2004; 131 (1): 117-130

    Abstract

    In both insects and mammals, olfactory receptor neurons (ORNs) expressing specific olfactory receptors converge their axons onto specific glomeruli, creating a spatial map in the brain. We have previously shown that second order projection neurons (PNs) in Drosophila are prespecified by lineage and birth order to send their dendrites to one of approximately 50 glomeruli in the antennal lobe. How can a given class of ORN axons match up with a given class of PN dendrites? Here, we examine the cellular and developmental events that lead to this wiring specificity. We find that, before ORN axon arrival, PN dendrites have already created a prototypic map that resembles the adult glomerular map, by virtue of their selective dendritic localization. Positional cues that create this prototypic dendritic map do not appear to be either from the residual larval olfactory system or from glial processes within the antennal lobe. We propose instead that this prototypic map might originate from both patterning information external to the developing antennal lobe and interactions among PN dendrites.

    View details for DOI 10.1242/dev.00896

    View details for Web of Science ID 000188553900012

    View details for PubMedID 14645123

  • Dendritic development of Drosophila high order visual system neurons is independent of sensory experience BMC NEUROSCIENCE Scott, E. K., Reuter, J. E., Luo, L. Q. 2003; 4

    Abstract

    The complex and characteristic structures of dendrites are a crucial part of the neuronal architecture that underlies brain function, and as such, their development has been a focal point of recent research. It is generally believed that dendritic development is controlled by a combination of endogenous genetic mechanisms and activity-dependent mechanisms. Therefore, it is of interest to test the relative contributions of these two types of mechanisms towards the construction of specific dendritic trees. In this study, we make use of the highly complex Vertical System (VS) of motion sensing neurons in the lobula plate of the Drosophila visual system to gauge the importance of visual input and synaptic activity to dendritic development.We find that the dendrites of VS1 neurons are unchanged in dark-reared flies as compared to control flies raised on a 12 hour light, 12 hour dark cycle. The dendrites of these flies show no differences from control in dendrite complexity, spine number, spine density, or axon complexity. Flies with genetically ablated eyes show a slight but significant reduction in the complexity and overall length of VS1 dendrites, although this effect may be due to a reduction in the overall size of the dendritic field in these flies.Overall, our results indicate no role for visual experience in the development of VS dendrites, while spontaneous activity from photoreceptors may play at most a subtle role in the formation of fully complex dendrites in these high-order visual processing neurons.

    View details for Web of Science ID 000185762300001

    View details for PubMedID 12834538

  • Axon pruning during Drosphila metamorphosis: Evidence for local degeneration and requirement of the ubiquitin-proteasome system NEURON Watts, R. J., Hoopfer, E. D., Luo, L. Q. 2003; 38 (6): 871-885

    Abstract

    Axon pruning is widely used for the refinement of neural circuits in both vertebrates and invertebrates, and may also contribute to the pathogenesis of neurodegenerative diseases. However, little is known about the cellular and molecular mechanisms of axon pruning. We use the stereotyped pruning of gamma neurons of the Drosophila mushroom bodies (MB) during metamorphosis to investigate these mechanisms. Detailed time course analyses indicate that MB axon pruning is mediated by local degeneration rather than retraction and that the disruption of the microtubule cytoskeleton precedes axon pruning. In addition, multiple lines of genetic evidence demonstrate an intrinsic role of the ubiquitin-proteasome system in axon pruning; for example, loss-of-function mutations of the ubiquitin activating enzyme (E1) or proteasome subunits in MB neurons block axon pruning. Our findings suggest that some forms of axon pruning during development may share similarities with degeneration of axons in response to injury.

    View details for Web of Science ID 000183691400008

    View details for PubMedID 12818174

  • Small GTPase Cdc42 is required for multiple aspects of dendritic morphogenesis JOURNAL OF NEUROSCIENCE Scott, E. K., Reuter, J. E., Luo, L. Q. 2003; 23 (8): 3118-3123

    Abstract

    The study of dendritic development in CNS neurons has been hampered by a lack of complex dendritic structures that can be studied in a tractable genetic system. In an effort to develop such a system, we recently characterized the highly complex dendrites of the vertical system (VS) neurons in the Drosophila visual system. Using VS neurons as a model system, we show here using loss-of-function mutations that endogenous Cdc42, a member of Rho family of small GTPases, is required for multiple aspects of dendritic morphogenesis. Cdc42-mutant VS neurons display normal complexity but increased dendritic length compared with wild type and have defects in dendrite caliber and stereotyped dendritic branch positions. Remarkably, Cdc42 mutant neurons also show a 50% reduction in dendritic spine density. These results demonstrate that Cdc42 is a regulator for multiple aspects of dendritic development.

    View details for Web of Science ID 000182475200005

    View details for PubMedID 12716918

  • A mosaic genetic screen for genes necessary for Drosophila mushroom body neuronal morphogenesis DEVELOPMENT Reuter, J. E., Nardine, T. M., Penton, A., Billuart, P., Scott, E. K., Usui, T., Uemura, T., Luo, L. Q. 2003; 130 (6): 1203-1213

    Abstract

    Neurons undergo extensive morphogenesis during development. To systematically identify genes important for different aspects of neuronal morphogenesis, we performed a genetic screen using the MARCM system in the mushroom body (MB) neurons of the Drosophila brain. Mutations on the right arm of chromosome 2 (which contains approximately 20% of the Drosophila genome) were made homozygous in a small subset of uniquely labeled MB neurons. Independently mutagenized chromosomes (4600) were screened, yielding defects in neuroblast proliferation, cell size, membrane trafficking, and axon and dendrite morphogenesis. We report mutations that affect these different aspects of morphogenesis and phenotypically characterize a subset. We found that roadblock, which encodes a dynein light chain, exhibits reduced cell number in neuroblast clones, reduced dendritic complexity and defective axonal transport. These phenotypes are nearly identical to mutations in dynein heavy chain Dhc64 and in Lis1, the Drosophila homolog of human lissencephaly 1, reinforcing the role of the dynein complex in cell proliferation, dendritic morphogenesis and axonal transport. Phenotypic analysis of short stop/kakapo, which encodes a large cytoskeletal linker protein, reveals a novel function in regulating microtubule polarity in neurons. MB neurons mutant for flamingo, which encodes a seven transmembrane cadherin, extend processes beyond their wild-type dendritic territories. Overexpression of Flamingo results in axon retraction. Our results suggest that most genes involved in neuronal morphogenesis play multiple roles in different aspects of neural development, rather than performing a dedicated function limited to a specific process.

    View details for DOI 10.1242/dev.00319

    View details for Web of Science ID 000181751300016

    View details for PubMedID 12571111

  • From lineage to wiring specificity: POU domain transcription factors control precise connections of Drosophila olfactory projection neurons CELL Komiyama, T., JOHNSON, W. A., Luo, L. Q., Jefferis, G. S. 2003; 112 (2): 157-167

    Abstract

    Axonal selection of synaptic partners is generally believed to determine wiring specificity in the nervous system. However, we have recently found evidence for specific dendritic targeting in the olfactory system of Drosophila: second order olfactory neurons (Projection Neurons) from the anterodorsal (adPN) and lateral (lPN) lineages send their dendrites to stereotypical, intercalating but non-overlapping glomeruli. Here we show that POU domain transcription factors, Acj6 and Drifter, are expressed in adPNs and lPNs respectively, and are required for their dendritic targeting. Moreover, misexpression of Acj6 in lPNs, or Drifter in adPNs, results in dendritic targeting to glomeruli normally reserved for the other PN lineage. Thus, Acj6 and Drifter translate PN lineage information into distinct dendritic targeting specificity. Acj6 also controls stereotypical axon terminal arborization of PNs in a central target, suggesting that the connectivity of PN axons and dendrites in different brain centers is coordinately regulated.

    View details for Web of Science ID 000181191600006

    View details for PubMedID 12553905

  • Structure of the vertical and horizontal system neurons of the lobula plate in Drosophila JOURNAL OF COMPARATIVE NEUROLOGY Scott, E. K., Raabe, T., Luo, L. Q. 2002; 454 (4): 470-481

    Abstract

    The lobula plate in the optic lobe of the fly brain is a high-order processing center for visual information. Within the lobula plate lie a small number of giant neurons that are responsible for the detection of wide field visual motion. Although the structure and motion sensitivity of these cells have been extensively described in large flies, the system has not been described systematically in Drosophila. Here, we use the mosaic analysis with a repressible cell marker (MARCM) system to analyze a subset of these cells, the horizontal and vertical systems. Our results suggest that the Drosophila horizontal system is similar to those described in larger flies, with three neurons fanning their dendrites over the lobula plate. We found that there are six neurons in the Drosophila vertical system, a figure that compares with 9-11 neurons in large flies. Even so, the Drosophila vertical system closely resembles the systems of larger flies, with each neuron in Drosophila having an approximate counterpart in large flies. This anatomical similarity implies that the inputs to the vertical system are similarly organized in these various fly species, and that it is likely that the Drosophila neurons respond to motions similar to those sensed by their specific structural counterparts in large flies. Additionally, the similar appearance of vertical system cells in multiple cell clones demonstrates that they share a common developmental lineage. Access to these cells in Drosophila should allow for the use of genetic tools in future studies of horizontal and vertical system function.

    View details for DOI 10.1002/cne.10467

    View details for Web of Science ID 000179558900008

    View details for PubMedID 12455010

  • Representation of the glomerular olfactory map in the Drosophila brain CELL Marin, E. C., Jefferis, G. S., Komiyama, T., Zhu, H. T., Luo, L. Q. 2002; 109 (2): 243-255

    Abstract

    We explored how the odor map in the Drosophila antennal lobe is represented in higher olfactory centers, the mushroom body and lateral horn. Systematic single-cell tracing of projection neurons (PNs) that send dendrites to specific glomeruli in the antennal lobe revealed their stereotypical axon branching patterns and terminal fields in the lateral horn. PNs with similar axon terminal fields tend to receive input from neighboring glomeruli. The glomerular classes of individual PNs could be accurately predicted based solely on their axon projection patterns. The sum of these patterns defines an "axon map" in higher olfactory centers reflecting which olfactory receptors provide input. This map is characterized by spatial convergence and divergence of PN axons, allowing integration of olfactory information.

    View details for Web of Science ID 000175082600013

    View details for PubMedID 12007410

  • Rac GTPases control axon growth, guidance and branching NATURE Ng, J., Nardine, T., Harms, M., Tzu, J., Goldstein, A., Sun, Y., Dietzl, G., Dickson, B. J., Luo, L. Q. 2002; 416 (6879): 442-447

    Abstract

    Growth, guidance and branching of axons are all essential processes for the precise wiring of the nervous system. Rho family GTPases transduce extracellular signals to regulate the actin cytoskeleton. In particular, Rac has been implicated in axon growth and guidance. Here we analyse the loss-of-function phenotypes of three Rac GTPases in Drosophila mushroom body neurons. We show that progressive loss of combined Rac1, Rac2 and Mtl activity leads first to defects in axon branching, then guidance, and finally growth. Expression of a Rac1 effector domain mutant that does not bind Pak rescues growth, partially rescues guidance, but does not rescue branching defects of Rac mutant neurons. Mosaic analysis reveals both cell autonomous and non-autonomous functions for Rac GTPases, the latter manifesting itself as a strong community effect in axon guidance and branching. These results demonstrate the central role of Rac GTPases in multiple aspects of axon development in vivo, and suggest that axon growth, guidance and branching could be controlled by differential activation of Rac signalling pathways.

    View details for Web of Science ID 000174607800050

    View details for PubMedID 11919635

  • Rac function and regulation during Drosophila development NATURE Hakeda-Suzuki, S., Ng, J., Tzu, J., Dietzl, G., Sun, Y., Harms, M., Nardine, T., Luo, L. Q., Dickson, B. J. 2002; 416 (6879): 438-442

    Abstract

    Rac GTPases regulate the actin cytoskeleton to control changes in cell shape. To date, the analysis of Rac function during development has relied heavily on the use of dominant mutant isoforms. Here, we use loss-of-function mutations to show that the three Drosophila Rac genes, Rac1, Rac2 and Mtl, have overlapping functions in the control of epithelial morphogenesis, myoblast fusion, and axon growth and guidance. They are not required for the establishment of planar cell polarity, as had been suggested on the basis of studies using dominant mutant isoforms. The guanine nucleotide exchange factor, Trio, is essential for Rac function in axon growth and guidance, but not for epithelial morphogenesis or myoblast fusion. Different Rac activators thus act in different developmental processes. The specific cellular response to Rac activation may be determined more by the upstream activator than the specific Rac protein involved.

    View details for Web of Science ID 000174607800049

    View details for PubMedID 11919634

  • Development of neuronal connectivity in Drosophila antennal lobes and mushroom bodies CURRENT OPINION IN NEUROBIOLOGY Jefferis, G. S., Marin, E. C., Watts, R. J., Luo, L. Q. 2002; 12 (1): 80-86

    Abstract

    Recent advances in the study of the connectivity of Drosophila olfactory system include the demonstration that olfactory receptor neurons project to specific glomeruli according to the receptor type they express, and that their projection neuron partners are prespecified to innervate particular glomeruli by birth order or time. This same theme of sequential generation has been observed in the generation of the three major types of mushroom body neurons.

    View details for Web of Science ID 000173813000010

    View details for PubMedID 11861168

  • Actin cytoskeleton regulation in neuronal morphogenesis and structural plasticity ANNUAL REVIEW OF CELL AND DEVELOPMENTAL BIOLOGY Luo, L. Q. 2002; 18: 601-635

    Abstract

    The actin cytoskeleton plays a major role in morphological development of neurons and in structural changes of adult neurons. This article reviews the myriad functions of actin and myosin in axon initiation, growth, guidance and branching, in morphogenesis of dendrites and dendritic spines, in synapse formation and stability, and in axon and dendrite retraction. Evidence is presented that signaling pathways involving the Rho family of small GTPases are key regulators of actin polymerization and myosin function in the context of different aspects of neuronal morphogenesis. These studies support an emerging theme: Different aspects of neuronal morphogenesis may involve regulation of common core signaling pathways, in particular the Rho GTPases.

    View details for DOI 10.1146/annurev.cellbio.18.031802.150501

    View details for Web of Science ID 000179413400022

    View details for PubMedID 12142283

  • Target neuron prespecification in the olfactory map of Drosophila NATURE Jefferis, G. S., Marin, E. C., Stocker, R. F., Luo, L. Q. 2001; 414 (6860): 204-208

    Abstract

    In Drosophila and mice, olfactory receptor neurons (ORNs) expressing the same receptors have convergent axonal projections to specific glomerular targets in the antennal lobe/olfactory bulb, creating an odour map in this first olfactory structure of the central nervous system. Projection neurons of the Drosophila antennal lobe send dendrites into glomeruli and axons to higher brain centres, thereby transferring this odour map further into the brain. Here we use the MARCM method to perform a systematic clonal analysis of projection neurons, allowing us to correlate lineage and birth time of projection neurons with their glomerular choice. We demonstrate that projection neurons are prespecified by lineage and birth order to form synapses with specific incoming ORN axons, and therefore to carry specific olfactory information. This prespecification could be used to hardwire the fly's olfactory system, enabling stereotyped behavioural responses to odorants. Developmental studies lead us to hypothesize that recognition molecules ensure reciprocally specific connections of ORNs and projection neurons. These studies also imply a previously unanticipated role for precise dendritic targeting by postsynaptic neurons in determining connection specificity.

    View details for Web of Science ID 000172029100047

    View details for PubMedID 11719930

  • Single neuron labeling and genetic manipulation NATURE NEUROSCIENCE Luo, L. Q., Zong, H. 2001; 4: 1158-1159

    View details for Web of Science ID 000172041500005

    View details for PubMedID 11687823

  • Regulating axon branch stability: The role of p190 RhoGAP in repressing a retraction signaling pathway CELL Billuart, P., Winter, C. G., Maresh, A., Zhao, X. S., Luo, L. Q. 2001; 107 (2): 195-207

    Abstract

    Mechanisms that regulate axon branch stability are largely unknown. Genome-wide analyses of Rho GTPase activating protein (RhoGAP) function in Drosophila using RNA interference identified p190 RhoGAP as essential for axon stability in mushroom body neurons, the olfactory learning and memory center. p190 inactivation leads to axon branch retraction, a phenotype mimicked by activation of GTPase RhoA and its effector kinase Drok and modulated by the level and phosphorylation of myosin regulatory light chain. Thus, there exists a retraction pathway from RhoA to myosin in maturing neurons, which is normally repressed by p190. Local regulation of p190 could control the structural plasticity of neurons. Indeed, genetic evidence supports negative regulation of p190 by integrin and Src, both implicated in neural plasticity.

    View details for Web of Science ID 000171694800009

    View details for PubMedID 11672527

  • Drosophila Rho-associated kinase (Drok) links frizzled-mediated planar cell polarity signaling to the actin cytoskeleton CELL Winter, C. G., Wang, B., Ballew, A., Royou, A., Karess, R., Axelrod, J. D., Luo, L. Q. 2001; 105 (1): 81-91

    Abstract

    Frizzled (Fz) and Dishevelled (Dsh) are components of an evolutionarily conserved signaling pathway that regulates planar cell polarity. How this signaling pathway directs asymmetric cytoskeletal reorganization and polarized cell morphology remains unknown. Here, we show that Drosophila Rho-associated kinase (Drok) works downstream of Fz/Dsh to mediate a branch of the planar polarity pathway involved in ommatidial rotation in the eye and in restricting actin bundle formation to a single site in developing wing cells. The primary output of Drok signaling is regulating the phosphorylation of nonmuscle myosin regulatory light chain, and hence the activity of myosin II. Drosophila myosin VIIA, the homolog of the human Usher Syndrome 1B gene, also functions in conjunction with this newly defined portion of the Fz/Dsh signaling pathway to regulate the actin cytoskeleton.

    View details for Web of Science ID 000168063300009

    View details for PubMedID 11301004

  • How do dendrites take their shape? NATURE NEUROSCIENCE Scott, E. K., Luo, L. Q. 2001; 4 (4): 359-365

    Abstract

    Recent technical advances have made possible the visualization and genetic manipulation of individual dendritic trees. These studies have led to the identification and characterization of molecules that are important for different aspects of dendritic development. Although much remains to be learned, the existing knowledge has allowed us to take initial steps toward a comprehensive understanding of how complex dendritic trees are built. In this review, we describe recent advances in our understanding of the molecular mechanisms underlying dendritic morphogenesis, and discuss their cell-biological implications.

    View details for Web of Science ID 000168762200015

    View details for PubMedID 11276225

  • enok encodes a Drosophila putative histone acetyltransferase required for mushroom body neuroblast proliferation CURRENT BIOLOGY Scott, E. K., Lee, T., Luo, L. Q. 2001; 11 (2): 99-104

    Abstract

    Mushroom bodies in the Drosophila brain are centers for olfactory learning and memory. We have previously shown that the mushroom bodies comprise three types of neurons with distinct axonal projections. These three types of neurons are generated sequentially from common neuroblasts. We report here the identification of a gene that we have named enoki mushroom (enok), which when it is mutated gives rise to mushroom bodies with reduced axonal structures. enok encodes a putative histone acetyltransferase (HAT) of the MYST family, members of which have been implicated as important modulators of transcriptional activity. A single amino acid change in the zinc finger motif of the putative catalytic HAT domain gives the same phenotype as a null allele, and this finding indicates the importance of HAT activity to Enok's function. Further phenotypic analysis demonstrates that the mushroom body defect is due to an arrest in neuroblast proliferation rather than a failure of either cell fate switching or axon branching. Clonal analyses in the wing discs and the ovaries suggest that enok is essential for normal cell proliferation in some, but not all, tissues. Our results provide in vivo evidence for essential functions of a histone acetyltransferase in the construction of the Drosophila brain.

    View details for Web of Science ID 000169076200018

    View details for PubMedID 11231125

  • Rho GTPases in neuronal morphogenesis NATURE REVIEWS NEUROSCIENCE Luo, L. Q. 2000; 1 (3): 173-180

    Abstract

    The Rho family of small GTPases act as intracellular molecular switches that transduce signals from extracellular stimuli to the actin cytoskeleton and the nucleus. Recent evidence implicates Rho GTPases in the regulation of neuronal morphogenesis, including migration, polarity, axon growth and guidance, dendrite elaboration and plasticity, and synapse formation. Signalling pathways from membrane receptors to Rho GTPases and from Rho GTPases to the actin cytoskeleton are beginning to be discovered. Mutations in these signalling pathways have been reported in human neurological diseases, which underscores their importance in the development and function of the nervous system.

    View details for Web of Science ID 000165764700015

    View details for PubMedID 11257905

  • Cell-autonomous requirement of the USP/EcR-B ecdysone receptor for mushroom body neuronal remodeling in Drosophila NEURON Lee, T., Marticke, S., Sung, C., Robinow, S., Luo, L. Q. 2000; 28 (3): 807-818

    Abstract

    Neuronal process remodeling occurs widely in the construction of both invertebrate and vertebrate nervous systems. During Drosophila metamorphosis, gamma neurons of the mushroom bodies (MBs), the center for olfactory learning in insects, undergo pruning of larval-specific dendrites and axons followed by outgrowth of adult-specific processes. To elucidate the underlying molecular mechanisms, we conducted a genetic mosaic screen and identified one ultraspiracle (usp) allele defective in larval process pruning. Consistent with the notion that USP forms a heterodimer with the ecdysone receptor (EcR), we found that the EcR-B1 isoform is specifically expressed in the MB gamma neurons, and is required for the pruning of larval processes. Surprisingly, most identified primary EcR/USP targets are dispensable for MB neuronal remodeling. Our study demonstrates cell-autonomous roles for EcR/USP in controlling neuronal remodeling, potentially through novel downstream targets.

    View details for Web of Science ID 000166057500018

    View details for PubMedID 11163268

  • Drosophila Lis1 is required for neuroblast proliferation, dendritic elaboration and axonal transport NATURE CELL BIOLOGY Liu, Z., Steward, R., Lu, L. Q. 2000; 2 (11): 776-783

    Abstract

    Haplo-insufficiency of human Lis1 causes lissencephaly. Reduced Lis1 activity in both humans and mice results in a neuronal migration defect. Here we show that Drosophila Lis1 is highly expressed in the nervous system. Lis1 is essential for neuroblast proliferation and axonal transport, as shown by a mosaic analysis using a Lis1 null mutation. Moreover, it is cell-autonomously required for dendritic growth, branching and maturation. Analogous mosaic analysis shows that neurons containing a mutated cytoplasmic-dynein heavy chain (Dhc64C) exhibit phenotypes similar to Lis1 mutants. These results implicate Lis1 as a regulator of the microtubule cytoskeleton and show that it is important for diverse physiological functions in the nervous system.

    View details for Web of Science ID 000165207500010

    View details for PubMedID 11056531

  • Small GTPases Rac and Rho in the maintenance of dendritic spines and branches in hippocampal pyramidal neurons JOURNAL OF NEUROSCIENCE Nakayama, A. Y., Harms, M. B., Luo, L. Q. 2000; 20 (14): 5329-5338

    Abstract

    The shape of dendritic trees and the density of dendritic spines can undergo significant changes during the life of a neuron. We report here the function of the small GTPases Rac and Rho in the maintenance of dendritic structures. Maturing pyramidal neurons in rat hippocampal slice culture were biolistically transfected with dominant GTPase mutants. We found that expression of dominant-negative Rac1 results in a progressive elimination of dendritic spines, whereas hyperactivation of RhoA causes a drastic simplification of dendritic branch patterns that is dependent on the activity of a downstream kinase ROCK. Our results suggest that Rac and Rho play distinct functions in regulating dendritic spines and branches and are vital for the maintenance and reorganization of dendritic structures in maturing neurons.

    View details for Web of Science ID 000087995300019

    View details for PubMedID 10884317

  • Trio quartet in D. (melanogaster) NEURON Luo, L. Q. 2000; 26 (1): 1-2

    View details for Web of Science ID 000086770500001

    View details for PubMedID 10798384

  • Essential roles of Drosophila RhoA in the regulation of neuroblast proliferation and dendritic but not axonal morphogenesis NEURON Lee, T. M., Winter, C., Marticke, S. S., Lee, A., Luo, L. Q. 2000; 25 (2): 307-316

    Abstract

    The pleiotropic functions of small GTPase Rho present a challenge to its genetic analysis in multicellular organisms. We report here the use of the MARCM (mosaic analysis with a repressible cell marker) system to analyze the function of RhoA in the developing Drosophila brain. Clones of cells homozygous for null RhoA mutations were specifically labeled in the mushroom body (MB) neurons of mosaic brains. We found that RhoA is required for neuroblast (Nb) proliferation but not for neuronal survival. Surprisingly, RhoA is not required for MB neurons to establish normal axon projections. However, neurons lacking RhoA overextend their dendrites, and expression of activated RhoA causes a reduction of dendritic complexity. Thus, RhoA is an important regulator of dendritic morphogenesis, while distinct mechanisms are used for axonal morphogenesis.

    View details for Web of Science ID 000085549100010

    View details for PubMedID 10719887

  • Intracellular signaling pathways that regulate dendritic spine morphogenesis HIPPOCAMPUS Nakayama, A. Y., Luo, L. Q. 2000; 10 (5): 582-586

    Abstract

    Rac is a member of the Rho family of small GTPases and acts as a molecular switch. When GTP-bound, Rac binds specific effectors to induce downstream signaling events, including actin cytoskeletal rearrangements (Hall, Science 1998;279:509-514). Herein we review the recent evidence suggesting that Rac is involved in the morphogenesis of dendritic spines (Luo et al., Nature 1996;379:837-840; Nakayama et al., J Neurosci 2000; 20:5329-5338). In addition, we discuss how Rac activity is regulated by guanine nucleotide exchange factors, which may be further regulated by extracellular factors. Thus, the Rac signal transduction pathway may provide links between extracellular ligands or synaptic activity and the regulation of the actin cytoskeleton in spine morphogenesis.

    View details for Web of Science ID 000165297700008

    View details for PubMedID 11075828

  • Development of the Drosophila mushroom bodies: sequential generation of three distinct types of neurons from a neuroblast DEVELOPMENT Lee, T., Lee, A., Luo, L. Q. 1999; 126 (18): 4065-4076

    Abstract

    The mushroom bodies (MBs) are prominent structures in the Drosophila brain that are essential for olfactory learning and memory. Characterization of the development and projection patterns of individual MB neurons will be important for elucidating their functions. Using mosaic analysis with a repressible cell marker (Lee, T. and Luo, L. (1999) Neuron 22, 451-461), we have positively marked the axons and dendrites of multicellular and single-cell mushroom body clones at specific developmental stages. Systematic clonal analysis demonstrates that a single mushroom body neuroblast sequentially generates at least three types of morphologically distinct neurons. Neurons projecting into the (gamma) lobe of the adult MB are born first, prior to the mid-3rd instar larval stage. Neurons projecting into the alpha' and beta' lobes are born between the mid-3rd instar larval stage and puparium formation. Finally, neurons projecting into the alpha and beta lobes are born after puparium formation. Visualization of individual MB neurons has also revealed how different neurons acquire their characteristic axon projections. During the larval stage, axons of all MB neurons bifurcate into both the dorsal and medial lobes. Shortly after puparium formation, larval MB neurons are selectively pruned according to birthdays. Degeneration of axon branches makes early-born gamma neurons retain only their main processes in the peduncle, which then project into the adult gamma lobe without bifurcation. In contrast, the basic axon projections of the later-born (alpha'/beta') larval neurons are preserved during metamorphosis. This study illustrates the cellular organization of mushroom bodies and the development of different MB neurons at the single cell level. It allows for future studies on the molecular mechanisms of mushroom body development.

    View details for Web of Science ID 000082965700009

    View details for PubMedID 10457015

  • Mosaic analysis with a repressible cell marker for studies of gene function in neuronal morphogenesis NEURON Lee, T., Luo, L. Q. 1999; 22 (3): 451-461

    Abstract

    We describe a genetic mosaic system in Drosophila, in which a dominant repressor of a cell marker is placed in trans to a mutant gene of interest. Mitotic recombination events between homologous chromosomes generate homozygous mutant cells, which are exclusively labeled due to loss of the repressor. Using this system, we are able to visualize axonal projections and dendritic elaboration in large neuroblast clones and single neuron clones with a membrane-targeted GFP marker. This new method allows for the study of gene functions in neuroblast proliferation, axon guidance, and dendritic elaboration in the complex central nervous system. As an example, we show that the short stop gene is required in mushroom body neurons for the extension and guidance of their axons.

    View details for Web of Science ID 000079483900010

    View details for PubMedID 10197526

  • Genghis Khan (Gek) as a putative effector for Drosophila Cdc42 and regulator of actin polymerization PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA Luo, L. Q., Lee, T., Tsai, L., Tang, G., Jan, L. Y., Jan, Y. N. 1997; 94 (24): 12963-12968

    Abstract

    The small GTPases Cdc42 and Rac regulate a variety of biological processes, including actin polymerization, cell proliferation, and JNK/mitogen-activated protein kinase activation, conceivably via distinct effectors. Whereas the effector for mitogen-activated protein kinase activation appears to be p65PAK, the identity of effector(s) for actin polymerization remains unclear. We have found a putative effector for Drosophila Cdc42, Genghis Khan (Gek), which binds to Dcdc42 in a GTP-dependent and effector domain-dependent manner. Gek contains a predicted serine/threonine kinase catalytic domain that is 63% identical to human myotonic dystrophy protein kinase and has protein kinase activities. It also possesses a large coiled-coil domain, a putative phorbol ester binding domain, a pleckstrin homology domain, and a Cdc42 binding consensus sequence that is required for its binding to Dcdc42. To study the in vivo function of gek, we generated mutations in the Drosophila gek locus. Egg chambers homozygous for gek mutations exhibit abnormal accumulation of F-actin and are defective in producing fertilized eggs. These phenotypes can be rescued by a wild-type gek transgene. Our results suggest that this multidomain protein kinase is an effector for the regulation of actin polymerization by Cdc42.

    View details for Web of Science ID A1997YJ45600050

    View details for PubMedID 9371783

Conference Proceedings


  • Cellular origins of wiring specificity in the olfactory system of Drosophila. Vyas, R. M., Jefferis, G., Berdnik, D., Ito, K., Luo, L. LIPPINCOTT WILLIAMS & WILKINS. 2004: S154-S154

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