Novel antibodies reveal presynaptic localization of C9orf72 protein and reduced protein levels in C9orf72 mutation carriers
In: Acta Neuropathologica Communications Acta Neuropathologica Communications, BioMed Central part of Springer Science, 2018, 6 (1), ⟨10.1186/s40478-018-0579-0⟩, Jg. 6 (2018), Heft 1, S. 1-17
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Zugriff:
Hexanucleotide repeat expansion in C9orf72 is the most common genetic cause of frontotemporal dementia and amyotrophic lateral sclerosis, but the pathogenic mechanism of this mutation remains unresolved. Haploinsufficiency has been proposed as one potential mechanism. However, insights if and how reduced C9orf72 proteins levels might contribute to disease pathogenesis are still limited because C9orf72 expression, localization and functions in the central nervous system (CNS) are uncertain, in part due to the poor specificity of currently available C9orf72 antibodies. Here, we generated and characterized novel knock-out validated monoclonal rat and mouse antibodies against C9orf72. We found that C9orf72 is a low abundant, cytoplasmic, highly soluble protein with the long 481 amino acid isoform being the predominant, if not exclusively, expressed protein isoform in mouse tissues and human brain. As consequence of the C9orf72 repeat expansion, C9orf72 protein levels in the cerebellum were reduced to 80% in our series of C9orf72 mutation carriers (n = 17) compared to controls (n = 26). However, no associations between cerebellar protein levels and clinical phenotypes were seen. Finally, by utilizing complementary immunohistochemical and biochemical approaches including analysis of human iPSC derived motor neurons, we identified C9orf72, in addition to its association to lysosomes, to be localized to the presynapses and able to interact with all members of the RAB3 protein family, suggestive of a role for C9orf72 in regulating synaptic vesicle functions by potentially acting as guanine nucleotide exchange factor for RAB3 proteins. In conclusion, our findings provide further evidence for haploinsufficiency as potential mechanism in C9orf72 pathogenesis by demonstrating reduced protein levels in C9orf72 mutation carriers and important novel insights into the physiological role of C9orf72 in the CNS. Moreover, the described novel monoclonal C9orf72 antibodies will be useful tools to further dissect the cellular and molecular functions of C9orf72. Electronic supplementary material The online version of this article (10.1186/s40478-018-0579-0) contains supplementary material, which is available to authorized users.
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Novel antibodies reveal presynaptic localization of C9orf72 protein and reduced protein levels in C9orf72 mutation carriers
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Autor/in / Beteiligte Person: | R. Jeroen Pasterkamp ; Neumann, Manuela ; Oulad-Abdelghani, Mustapha ; Cheng, Chieh-Yu ; Tahraoui-Bories, Julie ; Sellier, Chantal ; Feederle, Regina ; Frick, Petra ; Edbauer, Dieter ; Charlet-Berguerand, Nicolas ; Martinat, Cécile ; Mackenzie, Ian R. A. ; Prudlo, Johannes ; German Research Center for Neurodegenerative Diseases - Deutsches Zentrum für Neurodegenerative Erkrankungen (DZNE) ; Institut de Génétique et de Biologie Moléculaire et Cellulaire (IGBMC) ; Université de Strasbourg (UNISTRA)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS) ; University of British Columbia (UBC) ; Institut des cellules souches pour le traitement et l'étude des maladies monogéniques (I-STEM) ; Université d'Évry-Val-d'Essonne (UEVE)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Paris-Saclay-Généthon ; Association française contre les myopathies (AFM-Téléthon) ; Brain Centre Rudolf Magnus [Utrecht] ; University Medical Center [Utrecht] ; University of Rostock ; Munich Cluster for systems neurology [Munich] (SyNergy) ; Technische Universität Munchen - Université Technique de Munich [Munich, Allemagne] (TUM)-Ludwig-Maximilians-Universität München (LMU) ; Ludwig-Maximilians University [Munich] (LMU) ; Helmholtz-Zentrum München (HZM) ; University of Tübingen ; Ludwig-Maximilians-Universität München (LMU) ; Helmholtz Zentrum München = German Research Center for Environmental Health ; Technische Universität München [München] (TUM)-Ludwig-Maximilians-Universität München (LMU) ; CHARLET BERGUERAND, NICOLAS |
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Zeitschrift: | Acta Neuropathologica Communications Acta Neuropathologica Communications, BioMed Central part of Springer Science, 2018, 6 (1), ⟨10.1186/s40478-018-0579-0⟩, Jg. 6 (2018), Heft 1, S. 1-17 |
Veröffentlichung: | HAL CCSD, 2018 |
Medientyp: | unknown |
ISSN: | 2051-5960 (print) |
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