Alpha synuclein (alpha-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 alpha-syn aggregates suggested a role for alpha-syn in these disorders, although its biological activity remains poorly understood. Given the high degree of sequence similarity between vertebrate alpha-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 alpha-syn. The distribution of alpha-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 alpha-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 alpha-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 alpha-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 alpha-syn expression in cholinergic systems and suggest that alpha-syn modulates similar molecular pathways in phylogenetically distant vertebrates. (C) 2015 Wiley Periodicals, Inc.

Localization of α-synuclein in teleost central nervous system: immunohistochemical and Western blot evidence by 3D5 monoclonal antibody in the common carp, Cyprinus carpio

Vivacqua, Giorgio
Conceptualization
;
2015-01-01

Abstract

Alpha synuclein (alpha-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 alpha-syn aggregates suggested a role for alpha-syn in these disorders, although its biological activity remains poorly understood. Given the high degree of sequence similarity between vertebrate alpha-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 alpha-syn. The distribution of alpha-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 alpha-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 alpha-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 alpha-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 alpha-syn expression in cholinergic systems and suggest that alpha-syn modulates similar molecular pathways in phylogenetically distant vertebrates. (C) 2015 Wiley Periodicals, Inc.
5HT (RRID:AB_92263); ChAT (RRID:AB_2079760); TH (RRID:AB_390204); brain; spinal cord; Animals; Antibodies, Monoclonal; Antibody Specificity; Blotting, Western; Brain; Carps; Central Nervous System; Choline O-Acetyltransferase; Fluorescent Antibody Technique; Humans; Immunohistochemistry; Neurons; Serotonin; Species Specificity; Spinal Cord; Tyrosine 3-Monooxygenase; alpha-Synuclein
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/20.500.12610/70404
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