Localization of α-Synuclein in Teleost Central Nervous System: Immunohistochemical and Western Blot Evidence by 3D5 Monoclonal Antibody in the Common Carp, Cyprinus carpio

• Published in: Journal of comparative neurology (1911) 2015-05, Vol.523 (7), p.1095-1124
• Main Authors: Vaccaro, Rosa, Toni, Mattia, Casini, Arianna, Vivacqua, Giorgio, Yu, Shun, D'este, Loredana, Cioni, Carla
• Format: Article
• Language: English
• Subjects: 5HT (RRID:AB_92263); alpha-Synuclein - metabolism; Animals; Antibodies, Monoclonal - immunology; Antibody Specificity; Blotting, Western; brain; Brain - anatomy & histology; Brain - cytology; Carps - anatomy & histology; Central Nervous System - anatomy & histology; Central Nervous System - cytology; ChAT (RRID:AB_2079760); Choline O-Acetyltransferase - metabolism; Cyprinus carpio; Fluorescent Antibody Technique; Humans; Immunohistochemistry; Neurons - cytology; Neurons - metabolism; Serotonin - metabolism; Species Specificity; spinal cord; Spinal Cord - anatomy & histology; Teleostei; TH (RRID:AB_390204); Tyrosine 3-Monooxygenase - metabolism
• ISSN: 0021-9967; 1096-9861
• DOI: 10.1002/cne.23722

Alpha synuclein (α‐syn) is a 140 amino acid vertebrate‐specific protein, highly expressed in the human nervous system and abnormally accumulated in Parkinson's disease and other neurodegenerative disorders, known as synucleinopathies. The common occurrence of α‐syn aggregates suggested a role for α‐syn in these disorders, although its biological activity remains poorly understood. Given the high degree of sequence similarity between vertebrate α‐syns, we investigated this proteins in the central nervous system (CNS) of the common carp, Cyprinus carpio, with the aim of comparing its anatomical and cellular distribution with that of mammalian α‐syn. The distribution of α‐syn was analyzed by semiquantitative western blot, immunohistochemistry, and immunofluorescence by a novel monoclonal antibody (3D5) against a fully conserved epitope between carp and human α‐syn. The distribution of 3D5 immunoreactivity was also compared with that of choline acetyltransferase (ChAT), tyrosine hydroxylase (TH), and serotonin (5HT) by double immunolabelings. The results showed that a α‐syn‐like protein of about 17 kDa is expressed to different levels in several brain regions and in the spinal cord. Immunoreactive materials were localized in neuronal perikarya and varicose fibers but not in the nucleus. The present findings indicate that α‐syn‐like proteins may be expressed in a few subpopulations of catecholaminergic and serotoninergic neurons in the carp brain. However, evidence of cellular colocalization 3D5/TH or 3D5/5HT was rare. Differently, the same proteins appear to be coexpressed with ChAT by cholinergic neurons in several motor and reticular nuclei. These results sustain the functional conservation of the α‐syn expression in cholinergic systems and suggest that α‐syn modulates similar molecular pathways in phylogenetically distant vertebrates. J. Comp. Neurol. 523:1095–1124, 2015. © 2015 Wiley Periodicals, Inc. Using a novel monoclonal antibody (3D5) against α‐syn, the authors show the expression and the distribution of α‐syn like immunoreactivity in the carp central nervous system. In motor and reticular neurons α‐syn is clearly codistributed with ChAT. Therefore, the putative α‐syn positive neurons may represent a subpopulation of cholinergic neurons.