Endosomal location of dopamine receptors in neuronal cell cytoplasm |
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Authors: | Elizabeth C. Wolstencroft Goran Simic Nguyen thi Man Ian Holt Le Thanh Lam Paul R. Buckland Glenn E. Morris |
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Affiliation: | (1) Wolfson Centre for Inherited Neuromuscular Disease, RJAH Orthopaedic Hospital, LMARC Building, Oswestry, SY10 7AG, UK;(2) Institute of Science and Technology in Medicine, Keele University, Keele, ST4 7QB, UK;(3) Department of Neuroscience, Croatian Institute for Brain Research, University Medical School, Salata 12, Zagreb, Croatia;(4) Department of Psychological Medicine, University of Wales College of Medicine, Heath Park, Cardiff, UK;(5) Present address: Royal Devon and Exeter NHS Foundation Trust, Barrack Road, Exeter, EX2 5DW, UK |
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Abstract: | Five subtypes of dopamine receptor exist in two subfamilies: two D1-like (D1 and D5) and three D2-like (D2, D3 and D4). We produced novel monoclonal antibodies against all three D2-like receptors and used them to localize receptors in Ntera-2 (NT-2) cells, the human neuronal precursor cell line. Most of the immunostaining for all three receptors colocalized with mannose-6-phosphate receptor, a marker for late endosomes formed by internalization of the plasma membrane. This result was obtained with antibodies against three different epitopes on the D3 receptor, to rule out the possibility of cross-reaction with another protein, and controls without primary antibody or in the presence of competitor antigen were completely negative. In rat cerebral cortex and hippocampus, some of the dopamine receptor staining was found in similar structures in neuronal cell cytoplasm. Only some of the neurons were positive for dopamine receptors and the pattern was consistent with previously-reported patterns of innervation by dopamine-producing neurons. Endosomal dopamine receptors may provide a useful method for identifying cell bodies of dopamine-responsive neurons to complement methods that detect only active receptors in the neuronal cell membrane. |
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Keywords: | G protein-coupled receptor Endosome NT-2 cells Ntera-2 Monoclonal antibody Brain Hippocampus Cerebellum Cerebral cortex Mannose-6-phosphate receptor |
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