Bio

Honors & Awards


  • Rubicon grant - 24 months, Netherlands Organization for Scientific Research (2011)

Professional Education


  • Doctor of Philosophy, Universiteit Van Amsterdam (2010)

Stanford Advisors


Publications

Journal Articles


  • Glial fibrillary acidic protein isoform expression in plaque related astrogliosis in Alzheimer's disease NEUROBIOLOGY OF AGING Kamphuis, W., Middeldorp, J., Kooijman, L., Sluijs, J. A., Kooi, E., Moeton, M., Freriks, M., Mizee, M. R., Hol, E. M. 2014; 35 (3): 492-510

    Abstract

    In Alzheimer's disease (AD), amyloid plaques are surrounded by reactive astrocytes with an increased expression of intermediate filaments including glial fibrillary acidic protein (GFAP). Different GFAP isoforms have been identified that are differentially expressed by specific subpopulations of astrocytes and that impose different properties to the intermediate filament network. We studied transcript levels and protein expression patterns of all known GFAP isoforms in human hippocampal AD tissue at different stages of the disease. Ten different transcripts for GFAP isoforms were detected at different abundancies. Transcript levels of most isoforms increased with AD progression. GFAPδ-immunopositive astrocytes were observed in subgranular zone, hilus, and stratum-lacunosum-moleculare. GFAPδ-positive cells also stained for GFAPα. In AD donors, astrocytes near plaques displayed increased staining of both GFAPα and GFAPδ. The reading-frame-shifted isoform, GFAP(+1), staining was confined to a subset of astrocytes with long processes, and their number increased in the course of AD. In conclusion, the various GFAP isoforms show differential transcript levels and are upregulated in a concerted manner in AD. The GFAP(+1) isoform defines a unique subset of astrocytes, with numbers increasing with AD progression. These data indicate the need for future exploration of underlying mechanisms concerning the functions of GFAPδ and GFAP(+1) isoforms in astrocytes and their possible role in AD pathology.

    View details for DOI 10.1016/j.neurobiolaging.2013.09.035

    View details for Web of Science ID 000328655700007

    View details for PubMedID 24269023

  • GFAP delta Expression in Glia of the Developmental and Adolescent Mouse Brain PLOS ONE Mamber, C., Kamphuis, W., Haring, N. L., Peprah, N., Middeldorp, J., Hol, E. M. 2012; 7 (12)

    Abstract

    Glial fibrillary acidic protein (GFAP) is the major intermediate filament (IF) protein in astrocytes. In the human brain, GFAP isoforms have unique expression patterns, which indicate that they play distinct functional roles. One isoform, GFAP?, is expressed by proliferative radial glia in the developing human brain. In the adult human, GFAP? is a marker for neural stem cells. However, it is unknown whether GFAP? marks the same population of radial glia and astrocytes in the developing mouse brain as it does in the developing human brain. This study characterizes the expression pattern of GFAP? throughout mouse embryogenesis and into adolescence. Gfap? transcripts are expressed from E12, but immunohistochemistry shows GFAP? staining only from E18. This finding suggests a translational uncoupling. GFAP? expression increases from E18 to P5 and then decreases until its expression plateaus around P25. During development, GFAP? is expressed by radial glia, as denoted by the co-expression of markers like vimentin and nestin. GFAP? is also expressed in other astrocytic populations during development. A similar pattern is observed in the adolescent mouse, where GFAP? marks both neural stem cells and mature astrocytes. Interestingly, the Gfap?/Gfap? transcript ratio remains stable throughout development as well as in primary astrocyte and neurosphere cultures. These data suggest that all astroglia cells in the developing and adolescent mouse brain express GFAP?, regardless of their neurogenic capabilities. GFAP? may be an integral component of all mouse astrocytes, but it is not a specific neural stem cell marker in mice as it is in humans.

    View details for DOI 10.1371/journal.pone.0052659

    View details for Web of Science ID 000313158800086

    View details for PubMedID 23285135

  • GFAP in health and disease PROGRESS IN NEUROBIOLOGY Middeldorp, J., Hol, E. M. 2011; 93 (3): 421-443

    Abstract

    Glial fibrillary acidic protein (GFAP) is the main intermediate filament protein in mature astrocytes, but also an important component of the cytoskeleton in astrocytes during development. Major recent developments in astrocyte biology and the discovery of novel intermediate filament functions enticed the interest in the function of GFAP. The discovery of various GFAP splice variants gave an additional boost to explore this protein in more detail. The structural role of GFAP in astrocytes has been widely accepted for a long time, but over the years, GFAP has been shown to be involved in astrocyte functions, which are important during regeneration, synaptic plasticity and reactive gliosis. Moreover, different subpopulations of astrocytes have been identified, which are likely to have distinctive tasks in brain physiology and pathology, and which are not only classified by their spatial and temporal appearance, but also by their specific expression of intermediate filaments, including distinct GFAP isoforms. The presence of these isoforms enhances the complexity of the astrocyte cytoskeleton and is likely to underlie subtype specific functions. In this review we discuss the versatility of the GFAP cytoskeletal network from gene to function with a focus on astrocytes during human brain development, aging and disease.

    View details for DOI 10.1016/j.pneurobio.2011.01.005

    View details for Web of Science ID 000289339300006

    View details for PubMedID 21219963

  • Longterm quiescent cells in the aged human subventricular neurogenic system specifically express GFAP-delta AGING CELL van den Berge, S. A., Middeldorp, J., Zhang, C. E., Curtis, M. A., Leonard, B. W., Mastroeni, D., Voorn, P., van de Berg, W. D., Huitinga, I., Hol, E. M. 2010; 9 (3): 313-326

    Abstract

    A main neurogenic niche in the adult human brain is the subventricular zone (SVZ). Recent data suggest that the progenitors that are born in the human SVZ migrate via the rostral migratory stream (RMS) towards the olfactory bulb (OB), similar to what has been observed in other mammals. A subpopulation of astrocytes in the SVZ specifically expresses an assembly-compromised isoform of the intermediate filament protein glial fibrillary acidic protein (GFAP-delta). To further define the phenotype of these GFAP-delta expressing cells and to determine whether these cells are present throughout the human subventricular neurogenic system, we analysed SVZ, RMS and OB sections of 14 aged brain donors (ages 74-93). GFAP-delta was expressed in the SVZ along the ventricle, in the RMS and in the OB. The GFAP-delta cells in the SVZ co-expressed the neural stem cell (NSC) marker nestin and the cell proliferation markers proliferating cell nuclear antigen (PCNA) and Mcm2. Furthermore, BrdU retention was found in GFAP-delta positive cells in the SVZ. In the RMS, GFAP-delta was expressed in the glial net surrounding the neuroblasts. In the OB, GFAP-delta positive cells co-expressed PCNA. We also showed that GFAP-delta cells are present in neurosphere cultures that were derived from SVZ precursors, isolated postmortem from four brain donors (ages 63-91). Taken together, our findings show that GFAP-delta is expressed in an astrocytic subpopulation in the SVZ, the RMS and the OB. Importantly, we provide the first evidence that GFAP-delta is specifically expressed in longterm quiescent cells in the human SVZ, which are reminiscent of NSCs.

    View details for DOI 10.1111/j.1474-9726.2010.00556.x

    View details for Web of Science ID 000277789800003

    View details for PubMedID 20121722

  • Immunohistochemical characterization of the out-of frame splice variants GFAP Delta 164/Delta exon 6 in focal lesions associated with chronic epilepsy EPILEPSY RESEARCH Boer, K., Middeldorp, J., Spliet, W. G., Razavi, F., van Rijen, P. C., Baayen, J. C., Hol, E. M., Aronica, E. 2010; 90 (1-2): 99-109

    Abstract

    GFAP Delta164/Deltaexon 6 are two out-of frame splice variants of GFAP. The aim of this study was to investigate the distribution of GFAP Delta164/Deltaexon 6 expressing cells, in focal lesions associated with chronic intractable epilepsy, in light of the increasing interest in the role of specific astrocyte subtypes in epilepsy. Immunocytochemical analysis, using an antibody against Delta164 and Deltaexon6 (GFAP+1), was performed in surgical specimens of patients with hippocampal sclerosis (HS), focal cortical dysplasia type IIB (FCD), cortical tubers of tuberous sclerosis complex (TSC), glioneuronal and glial tumors. Expression of GFAP+1 was also evaluated in developing and adult human control cortex and hippocampus. GFAP+1 immunoreactivity was undetectable in developing human brain. In control human hippocampus and cortex (from young controls) only occasional GFAP+1 positive cells were observed. In contrast, GFAP+1 immunoreactivity was consistently detected in the glial component of the epileptogenic lesions. Balloon cells in FCD and giant cells in TSC, only rarely express GFAP+1. GFAP+1 co-localized with GFAPalpha, but not with GFAPdelta. Co-localization with aquaporin 4 was observed around blood vessels. GFAP+1 immunoreactivity in epilepsy-associated pathologies reveals a specific subpopulation of astrocytes in regions of astrogliosis. Further studies on GFAP+1 positive astrocytes are important to understand whether the expression of this isoform may affect the cytoskeletal integrity and the shape and function of glial cells under pathological conditions. However, while the staining is increased in epilepsy-associated pathologies, GFAP+1 is expressed in a small percentage of astrocytes. Thus, the possible role of this subpopulation of astrocytes in epilepsy is likely minor, compared to astrocytes expressing other GFAP isoforms.

    View details for DOI 10.1016/j.eplepsyres.2010.03.014

    View details for Web of Science ID 000279560700014

    View details for PubMedID 20430588

  • GFAP delta in radial glia and subventricular zone progenitors in the developing human cortex DEVELOPMENT Middeldorp, J., Boer, K., Sluijs, J. A., De Filippis, L., Encha-Razavi, F., Vescovi, A. L., Swaab, D. F., Aronica, E., Hol, E. M. 2010; 137 (2): 313-321

    Abstract

    A subpopulation of glial fibrillary acidic protein (GFAP)-expressing cells located along the length of the lateral ventricles in the subventricular zone (SVZ) have been identified as the multipotent neural stem cells of the adult mammalian brain. We have previously found that, in the adult human brain, a splice variant of GFAP, termed GFAPdelta, was expressed specifically in these cells. To investigate whether GFAPdelta is also present in the precursors of SVZ astrocytes during development and whether GFAPdelta could play a role in the developmental process, we analyzed GFAPdelta expression in the normal developing human cortex and in the cortex of foetuses with the migration disorder lissencephaly type II. We demonstrated for the first time that GFAPdelta is specifically expressed in radial glia and SVZ neural progenitors during human brain development. Expression of GFAPdelta in radial glia starts at around 13 weeks of pregnancy and disappears before birth. GFAPdelta is continuously expressed in the SVZ progenitors at later gestational ages and in the postnatal brain. Co-localization with Ki67 proved that these GFAPdelta-expressing cells are able to proliferate. Furthermore, we showed that the expression pattern of GFAPdelta was disturbed in lissencephaly type II. Overall, these results suggest that the adult SVZ is indeed a remnant of the foetal SVZ, which develops from radial glia. Furthermore, we provide evidence that GFAPdelta can distinguish resting astrocytes from proliferating SVZ progenitors.

    View details for DOI 10.1242/dev.041632

    View details for Web of Science ID 000273091500013

    View details for PubMedID 20040497

  • Specific Human Astrocyte Subtype Revealed by Affinity Purified GFAP(+1) Antibody; Unpurified Serum CrossReacts with Neurofilament-L in Alzheimer PLOS ONE Middeldorp, J., Van den Berge, S. A., Aronica, E., Speijer, D., Hol, E. M. 2009; 4 (11)

    Abstract

    The human GFAP splice variants GFAPDelta164 and GFAPDeltaexon6 both result in a GFAP protein isoform with a unique out-of-frame carboxy-terminus that can be detected by the GFAP+1 antibody. We previously reported that GFAP+1 was expressed in astrocytes and in degenerating neurons in Alzheimer's disease brains. In this study we aimed at further investigating the neuronal GFAP+1 expression and we started by affinity purifying the GFAP+1 antibody. The purified antibody resulted in a loss of neuronal GFAP+1 signal, although other antibodies directed against the amino- and carboxy-terminus of GFAPalpha still revealed GFAP-immunopositive neurons, as described before. With an in-depth analysis of a western blot, followed by mass spectrometry we discovered that the previously detected neuronal GFAP+1 expression was due to cross-reactivity of the antibody with neurofilament-L (NF-L). This was confirmed by double-label fluorescent immunohistochemistry and western blotting with the unpurified GFAP+1 antibody and an antibody against NF-L. Our data imply that NF-L can accumulate in some tangle-like structures in Alzheimer brains. More importantly, the purified GFAP+1 antibody clearly revealed a specific subtype of astrocytes in the adult human brain. These large astrocytes are present throughout the brain, e.g., along the subventricular zone, in the hippocampus, in the striatum and in the spinal cord of controls, Alzheimer, and Parkinson patients. The presence of a specific GFAP-isoform suggests a specialized function of these astrocytes.

    View details for DOI 10.1371/journal.pone.0007663

    View details for Web of Science ID 000271414600004

    View details for PubMedID 19888461

  • Intermediate filament transcription in astrocytes is repressed by proteasome inhibition FASEB JOURNAL Middeldorp, J., Kamphuis, W., Sluijs, J. A., Achoui, D., Leenaars, C. H., Feenstra, M. G., van Tijn, P., Fischer, D. F., Berkers, C., Ovaa, H., Quinlan, R. A., Hol, E. M. 2009; 23 (8): 2710-2726

    Abstract

    Increased expression of the astrocytic intermediate filament protein glial fibrillary acidic protein (GFAP) is a characteristic of astrogliosis. This process occurs in the brain during aging and neurodegeneration and coincides with impairment of the ubiquitin proteasome system. Inhibition of the proteasome impairs protein degradation; therefore, we hypothesized that the increase in GFAP may be the result of impaired proteasomal activity in astrocytes. We investigated the effect of proteasome inhibitors on GFAP expression and other intermediate filament proteins in human astrocytoma cells and in a rat brain model for astrogliosis. Extensive quantitative RT-PCR, immunocytochemistry, and Western blot analysis resulted unexpectedly in a strong decrease of GFAP mRNA to <4% of control levels [Control (DMSO) 100+/-19.2%; proteasome inhibitor (epoxomicin) 3.5+/-1.3%, n=8; P < or = 0.001] and a loss of GFAP protein in astrocytes in vitro. We show that the proteasome alters GFAP promoter activity, possibly mediated by transcription factors as demonstrated by a GFAP promoter-luciferase assay and RT(2) Profiler PCR array for human transcription factors. Most important, we demonstrate that proteasome inhibitors also reduce GFAP and vimentin expression in a rat model for induced astrogliosis in vivo. Therefore, proteasome inhibitors could serve as a potential therapy to modulate astrogliosis associated with CNS injuries and disease.

    View details for DOI 10.1096/fj.08-127696

    View details for Web of Science ID 000268836700039

    View details for PubMedID 19332645

  • Expression patterns of glial fibrillary acidic protein (GFAP)-delta in epilepsy-associated lesional pathologies NEUROPATHOLOGY AND APPLIED NEUROBIOLOGY Martinian, L., Boer, K., Middeldorp, J., Hol, E. M., Sisodiya, S. M., Squier, W., Aronica, E., Thom, M. 2009; 35 (4): 394-405

    Abstract

    Glial fibrillary acidic protein (GFAP)-delta is a novel isoform that differs in its C-terminal sequence from other GFAP isoforms. Previous studies suggest restriction of expression to the subpial layer, subventricular zone and the subgranular zone astrocytes, with an absence in pathological conditions causing reactive gliosis. GFAP-delta is speculated to have roles in regulation of astrocyte size and motility and a subpopulation of GFAP-delta-positive glia may be multipotent stem cells. The aim of this study was to investigate its expression in common causes of lesion-related refractory epilepsy.Hippocampal sclerosis (HS), focal cortical dysplasia (FCD) type IIB, cortical tuberous sclerosis (TSC) lesions, gangliogliomas, grey matter heterotopias and hemimegalencephaly from a wide age range of patients using both surgical and post mortem tissue specimens were studied.GFAP-delta expression was observed in CA4 and CA1 astrocytes in HS with less frequent labelling in the granule cell layer, even where granule cell dispersion was present. No significant labelling was noted in the subiculum in HS cases or in any subfields in non-HS epilepsy cases. Balloon cells in FCDIIB and hemimegalencephaly, giant cells in TSC and the astrocytic component of gangliogliomas showed immunoreactivity, colocalizing with conventional GFAP. No neuronal expression for GFAP-delta was seen in any of the pathologies. Quantitative analysis in 10 FCDIIB and five TSC cases revealed greater numbers of GFAP-delta-positive balloon cells than conventional GFAP. There was no GFAP-delta expression within nodular heterotopia.GFAP-delta expression patterns in HS overall appears to mirror regional reactive gliosis. It is a useful marker for the demonstration of balloon cells in FCD and TSC, which may be relevant to their abnormal size and localization. The lack of GFAP-delta within heterotopia supports their composition from cells destined for deeper cortical layers.

    View details for DOI 10.1111/j.1365-2990.2008.00996.x

    View details for Web of Science ID 000267711000005

    View details for PubMedID 19508443

  • Glial Fibrillary Acidic Protein Filaments Can Tolerate the Incorporation of Assembly-compromised GFAP-delta, but with Consequences for Filament Organization and alpha B-Crystallin Association MOLECULAR BIOLOGY OF THE CELL Perng, M., Wen, S., Gibbon, T., Middeldorp, J., Sluijs, J., Hol, E. M., Quinlan, R. A. 2008; 19 (10): 4521-4533

    Abstract

    The glial fibrillary acidic protein (GFAP) gene is alternatively spliced to give GFAP-alpha, the most abundant isoform, and seven other differentially expressed transcripts including GFAP-delta. GFAP-delta has an altered C-terminal domain that renders it incapable of self-assembly in vitro. When titrated with GFAP-alpha, assembly was restored providing GFAP-delta levels were kept low (approximately 10%). In a range of immortalized and transformed astrocyte derived cell lines and human spinal cord, we show that GFAP-delta is naturally part of the endogenous intermediate filaments, although levels were low (approximately 10%). This suggests that GFAP filaments can naturally accommodate a small proportion of assembly-compromised partners. Indeed, two other assembly-compromised GFAP constructs, namely enhanced green fluorescent protein (eGFP)-tagged GFAP and the Alexander disease-causing GFAP mutant, R416W GFAP both showed similar in vitro assembly characteristics to GFAP-delta and could also be incorporated into endogenous filament networks in transfected cells, providing expression levels were kept low. Another common feature was the increased association of alphaB-crystallin with the intermediate filament fraction of transfected cells. These studies suggest that the major physiological role of the assembly-compromised GFAP-delta splice variant is as a modulator of the GFAP filament surface, effecting changes in both protein- and filament-filament associations as well as Jnk phosphorylation.

    View details for DOI 10.1091/mbc.E08-03-0284

    View details for Web of Science ID 000260472200044

    View details for PubMedID 18685083

  • Ontogeny of hippocampal corticosteroid receptors: Effects of antenatal glucocorticoids in human and mouse JOURNAL OF COMPARATIVE NEUROLOGY Noorlander, C. W., de Graan, P. N., Middeldorp, J., van Beers, J. J., Visser, G. H. 2006; 499 (6): 924-932

    Abstract

    Women at risk for preterm delivery are treated with synthetic glucocorticoids (GCs) to enhance fetal lung maturation. GCs can bind to two intracellular receptors, the glucocorticoid receptor (GR) and the mineralocorticoid receptor (MR), which function as transcription factors. Both are highly expressed in the hippocampus. Several studies have focused on adverse side effects of antenatal GC treatment. However, relatively little is known about the ontogeny of GR and MR, especially in human. Therefore, we studied the ontogeny of both receptors in the human and mouse hippocampus and investigated the effects of antenatal dexamethasone (dex) treatment, a synthetic glucocorticoid, on MR and GR mRNA levels during hippocampal development. The results demonstrate that MR mRNA was first expressed in mouse hippocampus at embryonic day (E)15.5, at the timepoint when dex was administered. In contrast, GR mRNA expression was first observed after birth at postnatal day (P)5. However, in the human hippocampus both receptors are expressed at 24 weeks of gestation, when antenatal GCs are administered in clinical practice. Quantitative in situ hybridization demonstrated that MR mRNA levels were reduced only shortly after dex treatment at E16, but were unaffected from E18 onwards. These findings indicate that a single antenatal dex administration at E15.5 transiently affects MR mRNA levels in the mouse hippocampus. No effect of antenatal dex treatment was found on the human hippocampus at the third trimester of pregnancy. These data on the prenatal ontogeny of both corticosteroid receptors in the human hippocampus is important for understanding the significance of fetal glucocorticoid or stress exposure and its potential effects on health and disease.

    View details for DOI 10.1002/cne.21162

    View details for Web of Science ID 000242637500005

    View details for PubMedID 17072842

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