Bio

Academic Appointments


Professional Education


  • Postdoctoral, University of Pennsylvania, Biology, Physiology (1979)
  • PhD, Washington University School of Medicine, Yale University School of Medicine, Physiology and Biophysics (1978)
  • BSE, Princeton University, Electrical Engineering (1972)

Community and International Work


  • Squids4Kids, Pacific Grove, CA

    Topic

    Ocean health and marine ecology

    Partnering Organization(s)

    NOAA Southwest Fisheries Sciene Center, Monterey Bay National Marine Sancturary Foundation

    Location

    California

    Ongoing Project

    Yes

    Opportunities for Student Involvement

    Yes

  • Sustainable Utilization and Research of the Central Mar de Cortes (SURMAR), Santa Rosalia, BCS, Mexico

    Topic

    Marine ecology and resource develeopment

    Partnering Organization(s)

    Intsiuto Tecnologico Superior de Mulege, The Ocean Foundation

    Populations Served

    Mexican fishing communities and undergraduate students

    Location

    International

    Ongoing Project

    Yes

    Opportunities for Student Involvement

    Yes

Research & Scholarship

Current Research and Scholarly Interests


I've contributed to understanding electrical excitability of nerve & muscle in organisms ranging from brittle-stars to mammals, often using the squid giant axon as a model system. Currently working on behavior, physiology and ecology of Dosidicus gigas & Doryteuthis opalescens in the Gulf of CA & Monterey Bay. Electronic tagging & acoustic methods are used to track vertical & horizontal movements & to estimate biomass. Lab focuses on hypoxia tolerance, control of chromatophores & jet propulsion.

Teaching

2013-14 Courses


Graduate and Fellowship Programs


  • Biology (School of Humanities and Sciences) (Phd Program)

Publications

Journal Articles


  • Oceanographic and Biological Effects of Shoaling of the Oxygen Minimum Zone ANNUAL REVIEW OF MARINE SCIENCE, VOL 5 Gilly, W. F., Beman, J. M., Litvin, S. Y., Robison, B. H. 2013; 5: 393-420

    Abstract

    Long-term declines in oxygen concentrations are evident throughout much of the ocean interior and are particularly acute in midwater oxygen minimum zones (OMZs). These regions are defined by extremely low oxygen concentrations (<20-45 ?mol kg(-1)), cover wide expanses of the ocean, and are associated with productive oceanic and coastal regions. OMZs have expanded over the past 50 years, and this expansion is predicted to continue as the climate warms worldwide. Shoaling of the upper boundaries of the OMZs accompanies OMZ expansion, and decreased oxygen at shallower depths can affect all marine organisms through multiple direct and indirect mechanisms. Effects include altered microbial processes that produce and consume key nutrients and gases, changes in predator-prey dynamics, and shifts in the abundance and accessibility of commercially fished species. Although many species will be negatively affected by these effects, others may expand their range or exploit new niches. OMZ shoaling is thus likely to have major and far-reaching consequences.

    View details for DOI 10.1146/annurev-marine-120710-100849

    View details for Web of Science ID 000316390400018

    View details for PubMedID 22809177

  • Locomotion and behavior of Humboldt squid, Dosidicus gigas, in relation to natural hypoxia in the Gulf of California, Mexico JOURNAL OF EXPERIMENTAL BIOLOGY Gilly, W. F., Zeidberg, L. D., Booth, J. A., Stewart, J. S., Marshall, G., Abernathy, K., Bell, L. E. 2012; 215 (18): 3175-3190

    Abstract

    We studied the locomotion and behavior of Dosidicus gigas using pop-up archival transmitting (PAT) tags to record environmental parameters (depth, temperature and light) and an animal-borne video package (AVP) to log these parameters plus acceleration along three axes and record forward-directed video under natural lighting. A basic cycle of locomotor behavior in D. gigas involves an active climb of a few meters followed by a passive (with respect to jetting) downward glide carried out in a fins-first direction. Temporal summation of such climb-and-glide events underlies a rich assortment of vertical movements that can reach vertical velocities of 3 m s(-1). In contrast to such rapid movements, D. gigas spends more than 80% of total time gliding at a vertical velocity of essentially zero (53% at 0±0.05 m s(-1)) or sinking very slowly (28% at -0.05 to -0.15 m s(-1)). The vertical distribution of squid was compared with physical features of the local water column (temperature, oxygen and light). Oxygen concentrations of ?20 ?mol kg(-1), characteristic of the midwater oxygen minimum zone (OMZ), can influence the daytime depth of squid, but this depends on location and season, and squid can 'decouple' from this environmental feature. Light is also an important factor in determining daytime depth, and temperature can limit nighttime depth. Vertical velocities were compared over specific depth ranges characterized by large differences in dissolved oxygen. Velocities were generally reduced under OMZ conditions, with faster jetting being most strongly affected. These data are discussed in terms of increased efficiency of climb-and-glide swimming and the potential for foraging at hypoxic depths.

    View details for DOI 10.1242/jeb.072538

    View details for Web of Science ID 000308041400011

    View details for PubMedID 22915711

  • Diversity of conotoxin types from Conus californicus reflects a diversity of prey types and a novel evolutionary history TOXICON Elliger, C. A., Richmond, T. A., Lebaric, Z. N., PIERCE, N. T., Sweedler, J. V., Gilly, W. F. 2011; 57 (2): 311-322

    Abstract

    Most species within the genus Conus are considered to be specialists in their consumption of prey, typically feeding on molluscs, vermiform invertebrates or fish, and employ peptide toxins to immobilize prey. Conus californicus Hinds 1844 atypically utilizes a wide range of food sources from all three groups. Using DNA- and protein-based methods, we analyzed the molecular diversity of C. californicus toxins and detected a correspondingly large number of conotoxin types. We identified cDNAs corresponding to seven known cysteine-frameworks containing over 40 individual inferred peptides. Additionally, we found a new framework (22) with six predicted peptide examples, along with two forms of a new peptide type of unusual length. Analysis of leader sequences allowed assignment to known superfamilies in only half of the cases, and several of these showed a framework that was not in congruence with the identified superfamily. Mass spectrometric examination of chromatographic fractions from whole venom served to identify peptides corresponding to a number of cDNAs, in several cases differing in their degree of posttranslational modification. This suggests differential or incomplete biochemical processing of these peptides. In general, it is difficult to fit conotoxins from C. californicus into established toxin classification schemes. We hypothesize that the novel structural modifications of individual peptides and their encoding genes reflect evolutionary adaptation to prey species of an unusually wide range as well as the large phylogenetic distance between C. californicus and Indo-Pacific species.

    View details for DOI 10.1016/j.toxicon.2010.12.008

    View details for Web of Science ID 000287629400015

    View details for PubMedID 21172372

  • A diverse family of novel peptide toxins from an unusual cone snail, Conus californicus JOURNAL OF EXPERIMENTAL BIOLOGY Gilly, W. F., Richmond, T. A., Duda, T. F., Elliger, C., Lebaric, Z., Schulz, J., Bingham, J. P., Sweedler, J. V. 2011; 214 (1): 147-161

    Abstract

    Diversity among Conus toxins mirrors the high species diversity in the Indo-Pacific region, and evolution of both is thought to stem from feeding-niche specialization derived from intra-generic competition. This study focuses on Conus californicus, a phylogenetic outlier endemic to the temperate northeast Pacific. Essentially free of congeneric competitors, it preys on a wider variety of organisms than any other cone snail. Using molecular cloning of cDNAs and mass spectrometry, we examined peptides isolated from venom ducts to elucidate the sequences and post-translational modifications of two eight-cysteine toxins (cal12a and cal12b of type 12 framework) that block voltage-gated Na(+) channels. Based on homology of leader sequence and mode of action, these toxins are related to the O-superfamily, but differ significantly from other members of that group. Six of the eight cysteine residues constitute the canonical framework of O-members, but two additional cysteine residues in the N-terminal region define an O+2 classification within the O-superfamily. Fifteen putative variants of Cal12.1 toxins have been identified by mRNAs that differ primarily in two short hypervariable regions and have been grouped into three subtypes (Cal12.1.1-3). This unique modular variation has not been described for other Conus toxins and suggests recombination as a diversity-generating mechanism. We propose that these toxin isoforms show specificity for similar molecular targets (Na(+) channels) in the many species preyed on by C. californicus and that individualistic utilization of specific toxin isoforms may involve control of gene expression.

    View details for DOI 10.1242/jeb.046086

    View details for Web of Science ID 000285090000024

    View details for PubMedID 21147978

  • Two toxins from Conus striatus that individually induce tetanic paralysis BIOCHEMISTRY Kelley, W. P., Schulz, J. R., Jakubowski, J. A., Gilly, W. F., Sweedler, J. V. 2006; 45 (47): 14212-14222

    Abstract

    We describe structural properties and biological activities of two related O-glycosylated peptide toxins isolated from injected (milked) venom of Conus striatus, a piscivorous snail that captures prey by injecting a venom that induces a violent, spastic paralysis. One 30 amino acid toxin is identified as kappaA-SIVA (termed s4a here), and another 37 amino acid toxin, s4b, corresponds to a putative peptide encoded by a previously reported cDNA. We confirm the amino acid sequences and carry out structural analyses of both mature toxins using multiple mass spectrometric techniques. These include electrospray ionization ion-trap mass spectrometry and nanoelectrospray techniques for small volume samples, as well as matrix-assisted laser desorption/ionization time of flight mass spectrometric analysis as a complementary method to assist in the determination of posttranslational modifications, including O-linked glycosylation. Physiological experiments indicate that both s4a and s4b induce intense repetitive firing of the frog neuromuscular junction, leading to a tetanic contracture in muscle fiber. These effects apparently involve modification of voltage-gated sodium channels in motor axons. Notably, application of either s4a or s4b alone mimics the biological effects of the whole injected venom on fish prey.

    View details for DOI 10.1021/bi061485s

    View details for Web of Science ID 000242179100029

    View details for PubMedID 17115716

  • Intraspecific variation of venom injected by fish-hunting Conus snails JOURNAL OF EXPERIMENTAL BIOLOGY Jakubowski, J. A., Kelley, W. P., Sweedler, J. V., Gilly, W. F., Schulz, J. R. 2005; 208 (15): 2873-2883

    Abstract

    Venom peptides from two species of fish-hunting cone snails (Conus striatus and Conus catus) were characterized using microbore liquid chromatography coupled with matrix-assisted laser desorption/ionization-time of flight-mass spectrometry and electrospray ionization-ion trap-mass spectrometry. Both crude venom isolated from the venom duct and injected venom obtained by milking were studied. Based on analysis of injected venom samples from individual snails, significant intraspecific variation (i.e. between individuals) in the peptide complement is observed. The mixture of peptides in injected venom is simpler than that in the crude duct venom from the same snail, and the composition of crude venom is more consistent from snail to snail. While there is animal-to-animal variation in the peptides present in the injected venom, the composition of any individual's injected venom remains relatively constant over time in captivity. Most of the Conus striatus individuals tested injected predominantly a combination of two neuroexcitatory peptides (s4a and s4b), while a few individuals had unique injected-venom profiles consisting of a combination of peptides, including several previously characterized from the venom duct of this species. Seven novel peptides were also putatively identified based on matches of their empirically derived masses to those predicted by published cDNA sequences. Profiling injected venom of Conus catus individuals using matrix-assisted laser desorption/ionization-time of flight-mass spectrometry demonstrates that intraspecific variation in the mixture of peptides extends to other species of piscivorous cone snails. The results of this study imply that novel regulatory mechanisms exist to select specific venom peptides for injection into prey.

    View details for DOI 10.1242/jeb.01713

    View details for Web of Science ID 000231575800016

    View details for PubMedID 16043592

  • Decrease in inflammatory hyperalgesia by herpes vector-mediated knockdown of Na(v)1.7 sodium channels in primary afferents HUMAN GENE THERAPY Yeomans, D. C., Levinson, S. R., Peters, M. C., Koszowski, A. G., Tzabazis, A. Z., Gilly, W. F., Wilson, S. P. 2005; 16 (2): 271-277

    Abstract

    Induction of peripheral inflammation increases the expression of the Nav1.7 sodium channel in sensory neurons, potentially increasing their excitability. Peripheral inflammation also produces hyperalgesia in humans and an increase in nociceptive responsiveness in animals. To test the relationship between these two phenomena we applied a recombinant herpes simplex-based vector to the hindpaw skin of mice, which encoded both green fluorescent protein (GFP) as well as an antisense sequence to the Nav1.7 gene. The hindpaw was subsequently injected with complete Freund's adjuvant to induce robust inflammation. Application of the vector, but not a control vector encoding only GFP, prevented an increase in Nav1.7 expression in GFP-positive neurons and prevented development of hyperalgesia in both C and Adelta thermonociceptive tests. These results provide clear evidence of the involvement of an increased expression of the Nav1.7 channel in nociceptive neurons in the development of inflammatory hyperalgesia.

    View details for Web of Science ID 000227543900012

    View details for PubMedID 15761266

  • The projectile tooth of a fish-hunting cone snail: Conus catus injects venom into fish prey using a high-speed ballistic mechanism BIOLOGICAL BULLETIN Schulz, J. R., Norton, A. G., Gilly, W. F. 2004; 207 (2): 77-79

    View details for Web of Science ID 000224912700001

    View details for PubMedID 15501848

  • A gastropod toxin selectively slows early transitions in the Shaker K channel's activation pathway JOURNAL OF GENERAL PHYSIOLOGY Sack, J. T., Aldrich, R. W., Gilly, W. F. 2004; 123 (6): 685-696

    Abstract

    A toxin from a marine gastropod's defensive mucus, a disulfide-linked dimer of 6-bromo-2-mercaptotryptamine (BrMT), was found to inhibit voltage-gated potassium channels by a novel mechanism. Voltage-clamp experiments with Shaker K channels reveal that externally applied BrMT slows channel opening but not closing. BrMT slows K channel activation in a graded fashion: channels activate progressively slower as the concentration of BrMT is increased. Analysis of single-channel activity indicates that once a channel opens, the unitary conductance and bursting behavior are essentially normal in BrMT. Paralleling its effects against channel opening, BrMT greatly slows the kinetics of ON, but not OFF, gating currents. BrMT was found to slow early activation transitions but not the final opening transition of the Shaker ILT mutant, and can be used to pharmacologically distinguish early from late gating steps. This novel toxin thus inhibits activation of Shaker K channels by specifically slowing early movement of their voltage sensors, thereby hindering channel opening. A model of BrMT action is developed that suggests BrMT rapidly binds to and stabilizes resting channel conformations.

    View details for DOI 10.1085/jgp.200409047

    View details for Web of Science ID 000221988000006

    View details for PubMedID 15148327

  • Anatomical correlates of venom production in Conus californicus BIOLOGICAL BULLETIN Marshall, J., Kelley, W. P., Rubakhin, S. S., Bingham, J. P., Sweedler, J. V., Gilly, W. F. 2002; 203 (1): 27-41

    Abstract

    Like all members of the genus, Conus californicus has a specialized venom apparatus, including a modified radular tooth, with which it injects paralyzing venom into its prey. In this paper the venom duct and its connection to the pharynx, along with the radular sac and teeth, were examined using light and transmission electron microscopy. The general anatomy of the venom apparatus resembles that in other members of the genus, but several features are described that have not been previously reported for other species. The proximal (posterior) quarter of the venom duct is composed of a complex epithelium that may be specialized for active transport rather than secretion. The distal portion of the duct is composed of a different type of epithelium, suggestive of holocrine secretion, and the cells display prominent intracellular granules of at least two types. Similar granules fill the lumen of the duct. The passageway between the lumen of the venom duct and pharynx is a flattened branching channel that narrows to a width of 10 micro m and is lined by a unique cell type of unknown function. Granular material similar to that in the venom duct was also found in the lumen of individual teeth within the radular sac. Mass spectrometry (MALDI-TOF) demonstrated the presence of putative peptides in material derived from the tooth lumen, and all of the more prominent species were also evident in the anterior venom duct. Radular teeth thus appear to be loaded with peptide toxins while they are still in the radular sac.

    View details for Web of Science ID 000177717100003

    View details for PubMedID 12200253

  • Selective open-channel block of Shaker (Kv1) potassium channels by S-nitrosodithiothreitol (SNDTT) JOURNAL OF GENERAL PHYSIOLOGY Brock, M. W., Mathes, C., Gilly, W. F. 2001; 118 (1): 113-133

    Abstract

    Large quaternary ammonium (QA) ions block voltage-gated K(+) (Kv) channels by binding with a 1:1 stoichiometry in an aqueous cavity that is exposed to the cytoplasm only when channels are open. S-nitrosodithiothreitol (SNDTT; ONSCH(2)CH(OH)CH(OH)CH(2)SNO) produces qualitatively similar "open-channel block" in Kv channels despite a radically different structure. SNDTT is small, electrically neutral, and not very hydrophobic. In whole-cell voltage-clamped squid giant fiber lobe neurons, bath-applied SNDTT causes reversible time-dependent block of Kv channels, but not Na(+) or Ca(2)+ channels. Inactivation-removed ShakerB (ShBDelta) Kv1 channels expressed in HEK 293 cells are similarly blocked and were used to study further the action of SNDTT. Dose-response data are consistent with a scheme in which two SNDTT molecules bind sequentially to a single channel, with binding of the first being sufficient to produce block. The dissociation constant for the binding of the second SNDTT molecule (K(d2) = 0.14 mM) is lower than that of the first molecule (K(d1) = 0.67 mM), indicating cooperativity. The half-blocking concentration (K(1/2)) is approximately 0.2 mM. Steady-state block by this electrically neutral compound has a voltage dependence (about -0.3 e(0)) similar in magnitude but opposite in directionality to that reported for QA ions. Both nitrosyl groups on SNDTT (one on each sulfur atom) are required for block, but transfer of these reactive groups to channel cysteine residues is not involved. SNDTT undergoes a slow intramolecular reaction (tau approximately 770 s) in which these NO groups are liberated, leading to spontaneous reversal of the SNDTT effect. Competition with internal tetraethylammonium indicates that bath-applied SNDTT crosses the cell membrane to act at an internal site, most likely within the channel cavity. Finally, SNDTT is remarkably selective for Kv1 channels. When individually expressed in HEK 293 cells, rat Kv1.1-1.6 display profound time-dependent block by SNDTT, an effect not seen for Kv2.1, 3.1b, or 4.2.

    View details for Web of Science ID 000169782800010

    View details for PubMedID 11429448

  • Temperature-dependent expression of a squid Kv1 channel in Sf9 cells and functional comparison with the native delayed rectifier JOURNAL OF MEMBRANE BIOLOGY Brock, M. W., Lebaric, Z. N., Neumeister, H., Detomaso, A., Gilly, W. F. 2001; 180 (2): 147-161

    Abstract

    SqKv1A is a cDNA that encodes a Kv1 (Shaker-type) alpha-subunit expressed only in the giant axon and the parental giant fiber lobe (GFL) neurons of the squid stellate ganglion. We incorporated SqKv1A into a recombinant baculovirus for expression in the insect Sf9 cell line. Whole-cell patch-clamp recordings reveal that very few cells display functional potassium current (IK) if cultured at the standard postinfection temperature of 27 degrees C. At 18 degrees C, less SqKv1A protein is produced than at 27 degrees C, but cells with IK currents are much more numerous and can survive for at least 20 days postinfection (vs. approximately 5 days at 27 degrees C). Activation and deactivation kinetics of SqKv1A in Sf9 cells are slower (approximately 3- and 10-fold, respectively) than those of native channels in GFL neurons, but have similar voltage dependencies. The two cell types show only subtle differences in steady-state voltage-dependence of conductance and inactivation. Rates of IK inactivation in 20 mM external K are identical in the two cell types, but the sensitivity of inactivation to external tetraethylammonium (TEA) and K ions differ: inactivation of SqKv1A in Sf9 cells is slowed by external TEA and K ions, whereas inactivation of GFL IK is largely insensitive. Functional differences are discussed in terms of factors that may be specific to cell-type, including the presence of presently unidentified Kv1 subunits in GFL neurons that might form heteromultimers with SqKv1A.

    View details for Web of Science ID 000167640100005

    View details for PubMedID 11318098

  • Natural substitutions at highly conserved T1-domain residues perturb processing and functional expression of squid Kv1 channels JOURNAL OF NEUROPHYSIOLOGY Liu, T. I., Lebaric, Z. N., Rosenthal, J. J., Gilly, W. F. 2001; 85 (1): 61-71

    Abstract

    Shaker-type K-channel alpha-subunits (SqKv1A, B, D) expressed in neurons of the squid stellate ganglion differ in the length of their N-termini and in the species of amino acid present at several points in the T1 domain, an intracellular region involved in the tetramerization process during channel assembly. Heterologous expression of wild-type SqKv1A, B, and D in Xenopus oocytes reveals large differences in the level of both functional channels (assayed by whole-oocyte voltage clamp) and total channel protein (assayed by immunoblotting). Functional expression is poorest with SqKv1A and by far the best with SqKv1D. Biophysical properties of the three SqKv1 channels are essentially identical (assayed by cell-attached patch clamp). Site-directed mutagenesis was used to determine whether the observed differences in expression level are impacted by two residues in the T1 domain at which SqKv1A and B (but not D) differ from the consensus sequences found in many other taxa. In SqKv1A, glycine is substituted for arginine in an otherwise universally conserved sequence (FFDR in the T1(B) subdomain). In SqKv1B, glycine replaces serine in a sequence that is conserved within the Kv1 subfamily (SGLR in the T1(A) subdomain). Restoration of the consensus amino acid at these positions largely accounts for the observed differences in expression level. Analysis of the glycosylation state of aberrant versus restored alpha-subunits suggests that the anomalous amino acids in SqKv1A and B exert their influence during early steps in channel processing and assembly which take place in the endoplasmic reticulum (ER).

    View details for Web of Science ID 000166319300007

    View details for PubMedID 11152706

  • Effects of temperature on escape jetting in the squid Loligo opalescens JOURNAL OF EXPERIMENTAL BIOLOGY Neumeister, H., Ripley, B., Preuss, T., Gilly, W. F. 2000; 203 (3): 547-557

    Abstract

    In Loligo opalescens, a sudden visual stimulus (flash) elicits a stereotyped, short-latency escape response that is controlled primarily by the giant axon system at 15 C. We used this startle response as an assay to examine the effects of acute temperature changes down to 6 C on behavioral and physiological aspects of escape jetting. In free-swimming squid, latency, distance traveled and peak velocity for single escape jets all increased as temperature decreased. In restrained squid, intra-mantle pressure transients during escape jets increased in latency, duration and amplitude at low temperature. Recordings of stellar nerve activity revealed repetitive firing of the giant motor axon accompanied by increased activity in the non-giant motor axons that run in parallel. Selective stimulation of giant and non-giant motor axons in isolated nerve-muscle preparations failed to show the effects seen in vivo, i.e. increased peak force and increased neural activity at low temperature. Taken together, these results suggest that L. opalescens is able to compensate escape jetting performance for the effects of acute temperature reduction. A major portion of this compensation appears to occur in the central nervous system and involves alterations in the recruitment pattern of both the giant and non-giant axon systems.

    View details for Web of Science ID 000085498100012

    View details for PubMedID 10637183

  • Role of prey-capture experience in the development of the escape response in the squid Loligo opalescens: A physiological correlate in an identified neuron JOURNAL OF EXPERIMENTAL BIOLOGY Preuss, T., Gilly, W. F. 2000; 203 (3): 559-565

    Abstract

    Although extensively used for biophysical studies, the squid giant axon system remains largely unexplored in regard to in vivo function and modulation in any biologically relevant context. Here we show that successful establishment of the recruitment pattern for the giant axon in the escape response elicited by a brief electrical stimulus depends on prey-capture experience early in life. Juvenile squid fed only slow-moving, easy-to-capture prey items (Artemia salina) develop deficits in coordinating activity in the giant axon system with that of a parallel set of non-giant motor axons during escape responses. These deficits are absent in cohorts fed fast-moving, challenging prey items (copepods). These results suggest that the acquisition of inhibitory control over the giant axon system is experience-dependent and that both prey-capture and escape behavior depend on this control.

    View details for Web of Science ID 000085498100013

    View details for PubMedID 10637184

  • State-dependent nickel block of a high-voltage-activated neuronal calcium channel JOURNAL OF NEUROPHYSIOLOGY McFarlane, M. B., Gilly, W. F. 1998; 80 (4): 1678-1685

    Abstract

    Effect of nickel ions (Ni2+) on noninactivating calcium channels in squid giant fiber lobe (GFL) neurons were investigated with whole cell voltage clamp. Three different effects of Ni2+ were observed to be associated with distinct Ca2+ channel activation states. 1) Nickel ions appear to stabilize closed channel states and, as a result, slow activation kinetics. 2) Nickel ions block open channels with little voltage dependence over a wide range of potentials. 3) Block of open channels by Ni2+ becomes more effective during an extended strong depolarization, and this effect is voltage dependent. Recovery from this additional inhibition occurs at intermediate voltages, consistent with the presence of two distinct types of Ni2+ block that we propose correspond to two previously identified open states of the calcium channel. These results, taken together with earlier evidence of state-dependent block by omega-agatoxin IVA, suggest that Ni2+ generates these unique effects in part by interacting differently with the external surface of the GFL calcium channel complex in ways that depend on channel activation state.

    View details for Web of Science ID 000076487400007

    View details for PubMedID 9772231

  • A family of delayed rectifier Kv1 cDNAs showing cell type-specific expression in the squid stellate ganglion giant fiber lobe complex JOURNAL OF NEUROSCIENCE Rosenthal, J. J., Liu, T. I., Gilly, W. F. 1997; 17 (13): 5070-5079

    Abstract

    Squid giant axons are formed by giant fiber lobe (GFL) neurons of the stellate ganglion (SG). Other large motoneurons in the SG form a parallel system. A small family of cDNAs (SqKv1A-D) encoding Kv1 alpha-subunits was identified in a squid (Loligo opalescens) SG/GFL library. Members have distinct 5' untranslated regions (UTRs) and initial coding regions, but beyond a certain point (nucleotide 34 of SqKv1A) only nine differences exist. 3' UTRs are identical. Predicted alpha-subunits are nearly identical, and only the N termini differ significantly, primarily in length. RNase protection assays that use RNA isolated from specific SG regions show that SqKv1A mRNA is expressed prominently in the GFL but not in the SG proper. SqKv1B yields the opposite pattern. SqKv1D also is expressed only in the SG. SqKv1C expression was not detectable. In situ hybridizations confirm these results and reveal that SqKv1B mRNA is abundant in many large neurons of the SG, whereas SqKv1D expression is limited to small isolated clusters of neurons. SqKv1A and B are thus the predominant Kv1 mRNAs in the SG/GFL complex. Activation properties of SqKv1A and B channels expressed in oocytes are very similar to one another and compare favorably with properties of native delayed rectifier channels in GFL neurons and large SG neurons. The Kv1 complement in these squid neurons thus seems to be relatively simple. Several differences exist between cloned and native channels, however, and may reflect differences in the cellular environments of oocytes and neurons.

    View details for Web of Science ID A1997XE95200016

    View details for PubMedID 9185544

  • Post-hatching development of circular mantle muscles in the squid Loligo opalescens BIOLOGICAL BULLETIN Preuss, T., Lebaric, Z. N., Gilly, W. F. 1997; 192 (3): 375-387

    Abstract

    Post-hatching development of the circular muscles in the mantle of squid was studied morphometrically to identify structural changes and to quantify hyperplasia and hypertrophy of the muscle fibers. Superficial, mitochondria-rich (SMR) fibers and central, mitochondria-poor (CMP) fibers are present at hatching. Although both fiber types increase in size and, even more so, in number during post-hatching development, CMP fibers increase at a much higher rate than do SMR fibers. As a result, the relative proportion of SMR to CMP fibers shifts from about 1:1 in a hatchling to about 1:6 in an 8-week-old animal; it then apparently remains constant to adulthood. These structural changes are consistent with developmental changes in muscular activity. During slow, jet-propelled swimming, 1-week-old animals show mantle contractions that have twice the relative amplitude and frequency of those in adults. The presence of Na-channel protein in mantle muscle was detected bio-chemically by using site-directed antibodies; the protein was found to be preferentially expressed in CMP fibers. These results suggest that SMR fibers are an important source of locomotory power at hatching, but become progressively less important during the first 8 weeks of development as CMP fibers assume the dominant role in jet locomotion.

    View details for Web of Science ID A1997XH86200004

    View details for PubMedID 9212445

  • Fast and slow activation kinetics of voltage-gated sodium channels in molluscan neurons JOURNAL OF NEUROPHYSIOLOGY Gilly, W. F., Gillette, R., McFarlane, M. 1997; 77 (5): 2373-2384

    Abstract

    Whole cell patch-clamp recordings of Na current (I(Na)) were made under identical experimental conditions from isolated neurons from cephalopod (Loligo, Octopus) and gastropod (Aplysia, Pleurobranchaea, Doriopsilla) species to compare properties of activation gating. Voltage dependence of peak Na conductance (gNa) is very similar in all cases, but activation kinetics in the gastropod neurons studied are markedly slower. Kinetic differences are very pronounced only over the voltage range spanned by the gNa-voltage relation. At positive and negative extremes of voltage, activation and deactivation kinetics of I(Na) are practically indistinguishable in all species studied. Voltage-dependent rate constants underlying activation of the slow type of Na channel found in gastropods thus appear to be much more voltage dependent than are the equivalent rates in the universally fast type of channel that predominates in cephalopods. Voltage dependence of inactivation kinetics shows a similar pattern and is representative of activation kinetics for the two types of Na channels. Neurons with fast Na channels can thus make much more rapid adjustments in the number of open Na channels at physiologically relevant voltages than would be possible with only slow Na channels. This capability appears to be an adaptation that is highly evolved in cephalopods, which are well known for their high-speed swimming behaviors. Similarities in slow and fast Na channel subtypes in molluscan and mammalian neurons are discussed.

    View details for Web of Science ID A1997WZ56300011

    View details for PubMedID 9163364

  • Fast inactivation of delayed rectifier K conductance in squid giant axon and its cell bodies JOURNAL OF GENERAL PHYSIOLOGY Mathes, C., Rosenthal, J. J., Armstrong, C. M., Gilly, W. F. 1997; 109 (4): 435-448

    Abstract

    Inactivation of delayed rectifier K conductance (gk) was studied in squid giant axons and in the somata of giant fiber lobe (GFL) neurons. Axon measurements were made with an axial wire voltage clamp by pulsing to VK (approximately -10 mV in 50-70 mM external K) for a variable time and then assaying available gK with a strong, brief test pulse. GFL cells were studied with whole-cell patch clamp using the same prepulse procedure as well as with long depolarizations. Under our experimental conditions (12-18 degrees C, 4 mM internal MgATP) a large fraction of gK inactivates within 250 ms at -10 mV in both cell bodies and axons, although inactivation tends to be more complete in cell bodies. Inactivation in both preparations shows two kinetic components. The faster component is more temperature-sensitive and becomes very prominent above 12 degrees C. Contribution of the fast component to inactivation shows a similar voltage dependence to that of gK, suggesting a strong coupling of this inactivation path to the open state. Omission of internal MgATP or application of internal protease reduces the amount of fast inactivation. High external K decreases the amount of rapidly inactivating IK but does not greatly alter inactivation kinetics. Neither external nor internal tetraethylammonium has a marked effect on inactivation kinetics. Squid delayed rectifier K channels in GFL cell bodies and giant axons thus share complex fast inactivation properties that do not closely resemble those associated with either C-type or N-type inactivation of cloned Kvl channels studied in heterologous expression systems.

    View details for Web of Science ID A1997WT51900004

    View details for PubMedID 9101403

  • All-or-none contraction and sodium channels in a subset of circular muscle fibers of squid mantle BIOLOGICAL BULLETIN Gilly, W. F., Preuss, T., McFarlane, M. B. 1996; 191 (3): 337-340

    Abstract

    Motor function in squid (Loligo) mantle reflects the highly coordinated activity of two motor pathways associated with giant and non-giant motor axons that respectively produce all-or-none and graded contractions in mantle muscle. Whereas both types of axons innervate circular mantle muscle fibers, precise nerve-muscle relationships remain unclear. Are squid like most invertebrates, in which single muscle fibers receive dual innervation from giant and non-giant motor axons, or is squid mantle configured more like vertebrates, in which parallel motor axon systems innervate distinct fast and slow muscle fibers? In this report, we describe giant and nongiant motor pathways that appear to control different pools of circular muscle fibers in squid. A subset of circular muscle fibers possesses large Na currents, and these fibers are proposed to employ Na-dependent action potentials to produce fast, all-or-none muscle twitches associated with giant axon stimulation.

    View details for Web of Science ID A1996WA83400001

    View details for PubMedID 8976593

  • Molecular identification of SqKv1A - A candidate for the delayed rectifier K channel in squid giant axon JOURNAL OF GENERAL PHYSIOLOGY ROSENTHAL, J. C., Vickery, R. G., Gilly, W. F. 1996; 108 (3): 207-219

    Abstract

    We have cloned the cDNA for a squid Kvl potassium channel (SqKv1A). SqKv1A mRNA is selectively expressed in giant fiber lobe (GFL) neurons, the somata of the giant axons. Western blots detect two forms of SqKv1A in both GFL neuron and giant axon samples. Functional properties of SqKv1A currents expressed in Xenopus oocytes are very similar to macroscopic currents in GFL neurons and giant axons. Macroscopic K currents in GFL neuron cell bodies, giant axons, and in Xenopus oocytes expressing SqKv1A, activate rapidly and inactivate incompletely over a time course of several hundred ms. Oocytes injected with SqKv1A cRNA express channels of two conductance classes, estimated to be 13 and 20 pS in an internal solution containing 470 mM K. SqKv1A is thus a good candidate for the "20 pS" K channel that accounts for the majority of rapidly activating K conductance in both GFL neuron cell bodies and the giant axon.

    View details for Web of Science ID A1996VF64200008

    View details for PubMedID 8882864

  • Spatial localization of calcium channels in giant fiber lobe neurons of the squid (Loligo opalescens) PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA McFarlane, M. B., Gilly, W. F. 1996; 93 (10): 5067-5071

    Abstract

    Whole-cell voltage clamp was used to investigate the properties and spatial distribution of fast-deactivating (FD) Ca channels in squid giant fiber lobe (GFL) neurons. Squid FD Ca channels are reversibly blocked by the spider toxin omega-Agatoxin IVA with an IC50 of 240-420 nM with no effect on the kinetics of Ca channel gating. Channels with very similar properties are expressed in both somatic and axonal domains of cultured GFL neurons, but FD Ca channel conductance density is higher in axonal bulbs than in cell bodies at all times in culture. Channels presumably synthesized during culture are preferentially expressed in the growing bulbs, but bulbar Ca conductance density remains constant while Na conductance density increases, suggesting that processes determining the densities of Ca and Na channels in this extrasomatic domain are largely independent. These observations suggest that growing axonal bulbs in cultured GFL neurons are not composed entirely of "axonal" membranes because FD Ca channels are absent from the giant axon in situ but, rather, suggest a potential role for FD Ca channels in mediating neurotransmitter release at the motor terminals of the giant axon.

    View details for Web of Science ID A1996UL25500100

    View details for PubMedID 8643530

  • Methadone block of K+ current in squid giant fiber lobe neurons JOURNAL OF GENERAL PHYSIOLOGY Horrigan, F. T., Gilly, W. F. 1996; 107 (2): 243-260

    Abstract

    Voltage-dependent ionic currents were recorded from squid giant fiber lobe neurons using the whole-cell patch-clamp technique. When applied to the bathing solution, methadone was found to block IK, I Na and I Ca. Both I Na and I Ca were reduced without apparent change in kinetics and exhibited IC(50)'s of 50-100 and 250-500 mu M, respectively, at +10 mV. In contrast, IK was reduced in a time-dependent manner that is well fit by a simple model of open channel block (K(D)= 32+/- or 2 mu M, +60 mV, 10 degrees Celsius). The mechanism of I(K) block was examined in detail and involves a direct action of methadone, a tertiary amine, on K channels rather than an opioid receptor-mediated pathway. The kinetics of I(K) block resemble those reported for internally applied long chain quaternary ammonium (QA) compounds; and recovery from I(K) block is QA-like in its slow time course and strong dependence on holding potential. A quaternary derivative of methadone (N-methyl-methadone) only reproduced the effects of methadone on I(K) when included in the pipette solution; this compound was without effect when applied externally. I(K) block thus appears to involve diffusion of methadone into the cytoplasm and occlusion of the open K channel at the internal QA blocking site by the protonated form of the drug. This proposed mode of action is supported by the pH and voltage dependence of block as well as by the observation that high external K+ speeds the rate of drug dissociation. In addition, the effect of methadone on I(K) evoked during prolonged (300 ms) depolarizations suggests that methadone block may interfere with endogenous K+ channel inactivation. The effects of temperature, methadone stereoisomers, and the methadone-like drugs propoxyphene and nor-propoxyphene on IK block were examined. Methadone was also found to block I(K) in GH3 cells and in chick myoblasts.

    View details for Web of Science ID A1996TY12500007

    View details for PubMedID 8833344

  • Ontogeny of copepod predation in juvenile squid (Loligo opalescens) BIOLOGICAL BULLETIN Chen, D. S., VanDykhuizen, G., Hodge, J., Gilly, W. F. 1996; 190 (1): 69-81

    Abstract

    Copepods are the major prey of juvenile squid, and small species of squid such as Loligo opalescens face a great challenge in catching these erratically moving crustaceans. We studied the ontogeny of copepod predation in laboratory-reared animals and found that mastery of copepod capture develops progressively, starting shortly after hatch with strong attacks of a simple type. Modifications of the initial basic attack lead to more specialized strategies that effectively extend the range of capture to both longer and shorter distances. This progression culminates, by approximately 40 days post-hatching, in adult-like prey capture behavior involving tentacle extension and retraction. Squid raised exclusively on easily captured Artemia nauplii and introduced to a copepod diet 40 days after hatching displayed only basic attack behavior, characteristic of very young squid. All of these attacks were unsuccessful, and very few of these animals survived the transition. Copepod capture thus appears to be a skill that must be acquired in an experience-dependent manner early in post-hatching life.

    View details for Web of Science ID A1996TY80400008

    View details for PubMedID 8852631

  • Tissue distribution and subcellular localization of Na+ channel mRNA in the nervous system of the squid, Loligo opalescens RECEPTORS & CHANNELS Liu, T. I., Gilly, W. F. 1995; 3 (4): 243-?

    Abstract

    Recent cloning of a putative Na+ channel alpha subunit cDNA, GFLN1, from the squid stellate ganglion has allowed us to study the expression of this ion channel at a cellular level. In situ hybridizations with a probe derived from and specific to 3' untranslated and coding sequence of GFLN1 were used to determine its tissue distribution as well as its subcellular localization. In sections of the stellate ganglion, the probe labeled all of the cells in the giant fiber lobe (GFL) and most cells in the cellular layer of the main ganglion. In these non-GFL portions of the stellate ganglion, labeling was particularly intense in the ventral large cells and weak or absent in the dorsal small cells. In the optic lobe, only a select group of cells, the second-order visual giant neurons, were intensely labeled. These results are consistent with electrophysiological data that show GFL-like Na+ currents in rare large cells dissociated from the optic lobe and in most but not all cells from the non-GFL part of the stellate ganglion. In sections of the subesophageal mass of the central nervous system, strong labeling for GFLN1 mRNA occurred in the fin lobe, posterior chromatophore lobe, central and latero-ventral palliovisceral lobes, and posterior pedal lobe. In all cases, labeling was detected only in the cellular layer of these tissues and never in nerves or neuropil. In situ hybridization with dissociated GFL neurons maintained in primary culture verified that Na+ channel mRNA is confined to the cell body. These results indicate that GFLN1 is expressed predominately in large cells with large or long axons, and that this mRNA is restricted to the cell bodies of these neurons.

    View details for Web of Science ID A1995UG60900001

    View details for PubMedID 8833997

  • AMINO-ACID-SEQUENCE OF A PUTATIVE SODIUM-CHANNEL EXPRESSED IN THE GIANT-AXON OF THE SQUID LOLIGO-OPALESCENS PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA Rosenthal, J. J., Gilly, W. F. 1993; 90 (21): 10026-10030

    Abstract

    A full-length cDNA encoding a putative Na+ channel (GFLN1) has been cloned from a library prepared from the stellate ganglion of Loligo opalescens. The cDNA encodes a predicted protein of 1784 amino acids. Regions of the GFLN1 protein with defined functional importance (membrane span S4, the SS1 and SS2 segments, and interdomain III-IV) are highly conserved among all vertebrate Na+ channel alpha-subunit structures. Northern blot hybridization and RNase protection assays verify that mRNA corresponding to GFLN1 is expressed in neurons of the giant fiber lobe that form the giant axon. We propose that GFLN1 encodes the Na+ channel that has been extensively studied in the squid axon.

    View details for Web of Science ID A1993MF29600059

    View details for PubMedID 8234251

  • VOLTAGE-DEPENDENT CALCIUM AND POTASSIUM CONDUCTANCES IN STRIATED-MUSCLE FIBERS FROM THE SCORPION, CENTRUROIDES-SCULPTURATUS JOURNAL OF MEMBRANE BIOLOGY Gilly, W. F., Scheuer, T. 1993; 134 (2): 155-167

    Abstract

    Ionic currents responsible for the action potential in scorpion muscle fibers were characterized using a three-intracellular microelectrode voltage clamp applied at the fiber ends (8-12 degrees C). Large calcium currents (ICa) trigger contractile activation in physiological saline (5 mM Ca) but can be studied in the absence of contractile activation in a low Ca saline (< or = 2.5 mM). Barium (Ba) ions (1.5-3 mM) support inward current but not contractile activation. Ca conductance kinetics are fast (time constant of 3 msec at 0 mV) and very voltage dependent, with steady-state conductance increasing e-fold in approximately 4 mV. Half-activation occurs at -25 mV. Neither ICa nor IBa show rapid inactivation, but a slow, voltage-dependent inactivation eliminates ICa at voltages positive to -40 mV. Kinetically, scorpion channels are more similar to L-type Ca channels in vertebrate cardiac muscle than to those in skeletal muscle. Outward K currents turn on more slowly and with a longer delay than do Ca currents, and K conductance rises less steeply with voltage (e-fold change in 10 mV; half-maximal level at 0 mV). K channels are blocked by externally applied tetraethylammonium and 3,4 diaminopyridine.

    View details for Web of Science ID A1993LH16000007

    View details for PubMedID 8411118

  • ELECTRICAL RESPONSES TO CHEMICAL-STIMULATION OF SQUID OLFACTORY RECEPTOR-CELLS JOURNAL OF EXPERIMENTAL BIOLOGY Lucero, M. T., Horrigan, F. T., Gilly, W. F. 1992; 162: 231-249
  • CONTROL OF THE SPATIAL-DISTRIBUTION OF SODIUM-CHANNELS IN GIANT FIBER LOBE NEURONS OF THE SQUID NEURON Gilly, W. F., Lucero, M. T., Horrigan, F. T. 1990; 5 (5): 663-674

    Abstract

    Na+ channels are present at high density in squid giant axon but are absent from its somata in the giant fiber lobe (GFL) of the stellate ganglion. GFL cells dispersed in vitro maintain growing axons and develop a Na+ channel distribution similar to that in vivo. Tunicamycin, a glycosylation inhibitor, selectively disrupts the spatially appropriate, high level expression of Na+ channels in axonal membrane but has no effect on expression in cell bodies, which show low level, inappropriate expression in vitro. This effect does not appear to involve alteration in Na+ channel turnover or axon viability. K+ channel distribution is unaffected. Thus, glycosylation appears to be involved in controlling Na+ channel localization in squid neurons.

    View details for Web of Science ID A1990EJ86800010

    View details for PubMedID 2171590

  • JET-PROPELLED ESCAPE IN THE SQUID LOLIGO-OPALESCENS - CONCERTED CONTROL BY GIANT AND NON-GIANT MOTOR AXON PATHWAYS PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA Otis, T. S., Gilly, W. F. 1990; 87 (8): 2911-2915

    Abstract

    Recordings of stellar nerve activity were made during escape responses in living squid. Short-latency activation of the giant axons is triggered by light-flash stimulation that elicits a stereotyped startle-escape response and powerful jet. Many other types of stimuli produce a highly variable, delayed-escape response with strong jetting primarily controlled by a small axon motor pathway. In such cases, activation of the giant axons is not necessary for a vigorous escape jet. When they are utilized, the giant axons are not activated until well after the non-giant system initiates the escape response, and excitation is critically timed to boost the rise in intramantle pressure. Squid thus show at least two escape modes in which the giant axons can contribute in different ways to the control of a highly flexible behavior.

    View details for Web of Science ID A1990CZ29900010

    View details for PubMedID 2326255

  • PROPERTIES OF APPROPRIATELY AND INAPPROPRIATELY EXPRESSED SODIUM-CHANNELS IN SQUID GIANT-AXON AND ITS SOMATA JOURNAL OF NEUROSCIENCE Gilly, W. F., Brismar, T. 1989; 9 (4): 1362-1374

    Abstract

    Neurons that form the giant axons in squid by axonal fusion in the stellate ganglion are inexcitable and do not express functional voltage-controlled sodium (Na) channels in their somata in vivo. These cells do express Na channels in the soma membrane in vitro, however, provided they have been axotomized. We describe here voltage-clamp experiments on the isolated cell bodies maintained in primary culture and on acutely isolated giant axons designed to compare the functional properties of the Na channels expressed inappropriately in the soma with those of channels expressed normally in the axon. Approximately 85% of Na channels in the soma are essentially indistinguishable from those in the giant axon with regard to gating properties and sensitivity to tetrodotoxin or saxitoxin. Thus, the isolated soma is capable of processing Na channels to a state of apparent functional perfection. In addition to these normal Na channels, another type is regularly expressed in the cultured somata. This second type lacks inactivation and is preferentially sensitive to block by cadmium ions, but is otherwise indistinguishable from the more prevalent normal type of channels.

    View details for Web of Science ID A1989U297500024

    View details for PubMedID 2539444

  • PHYSIOLOGICAL-PROPERTIES OF 3 MUSCLE-FIBER TYPES CONTROLLING DORSAL FIN MOVEMENTS IN A FLATFISH, CITHARICHTHYS-SORDIDUS JOURNAL OF MUSCLE RESEARCH AND CELL MOTILITY Gilly, W. F., ALADJEM, E. 1987; 8 (5): 407-417

    Abstract

    Pacific sand dabs utilize their dorsal and anal fins in different behaviours which are characterized by extremely rapid fin movements, on one hand, and essentially isometric force generation on the other. Muscle fibres controlling fin movements were examined physiologically. Direct electrical shocks to localized regions of fin muscles reveal three fibre types. The longest fibres in the muscle are very fast and functionally analogous to frog twitch fibres. The shortest fibres are extremely, slow and show properties much like frog tonic fibres. The mid-length fibres produce contractile responses which are intermediate in time course. Even the fastest muscle fibres do not generate action potentials, but instead rely on summating junction potentials to drive membrane voltage to a stable level just beyond contraction threshold (-35 to -40 mV). Twitch amplitude can be finely graded by the time that membrane depolarization exceeds this threshold level.

    View details for Web of Science ID A1987L073000004

    View details for PubMedID 3429642

  • SYNTHESIS OF SODIUM-CHANNELS IN THE CELL-BODIES OF SQUID GIANT-AXONS PROCEEDINGS OF THE NATIONAL ACADEMY OF SCIENCES OF THE UNITED STATES OF AMERICA Brismar, T., Gilly, W. F. 1987; 84 (5): 1459-1463

    Abstract

    Giant axons in squid are formed by fusion of axons from many small cell bodies in the giant fiber lobe (GFL) of the stellate ganglion. Somata of GFL cells in vivo are inexcitable and do not have measurable sodium current (INa) when studied with microelectrode or patch-electrode voltage-clamp techniques. If GFL cells are separated from the giant axons and maintained in primary culture, axon-like INa can be recorded from the somata after several days. Incorporation of Na channels into GFL cell bodies requires protein synthesis, intracellular microtubule-based transport, and the lack of a morphologically defined axon to serve as a sink for channels synthesized in culture.

    View details for Web of Science ID A1987G304200072

    View details for PubMedID 3469679

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