Dopamine D5 receptor immunolocalization in rat and monkey brain

Dopamine D5 receptor localization has been difficult because even the most specific ligands cannot distinguish between molecular subtypes of the D1‐like receptor subfamily. Antifusion protein rabbit polyclonal antibodies directed against the C‐terminus of human D5 receptor were therefore developed for immunolocalization of the D5 receptor protein in brain. The antibodies were characterized by immunoblot analysis and immunoprecipitation and used for light microscopic immunocytochemistry in rat and monkey brain. Affinity purified D5 antibodies were specific for D5 fusion protein as well as cloned and native D5 receptor on Western blots, and D5 antisera specifically immunoprecipitated solubilized, cloned D5 receptor. Regional distribution of D5 receptor immunoreactivity was consistent across species and correlated well with D5 mRNA distribution previously reported in monkey brain. Immunoreactivity was widespread and tended to label perikarya and proximal dendrites of neurons in cerebral cortex, basal ganglia, basal forebrain, hippocampus, diencephalon, brainstem, and cerebellum. Neuropil was immunoreactive in olfactory bulb, islands of Calleja, cerebral cortex, superior colliculus, and molecular layer of cerebellum. The distribution of D5 in brain was clearly different from that of other dopamine receptor subtypes, including D1, the other member of the D1‐like receptor subfamily. This unique distribution corroborates the idea that the D5 receptor subtype has a distinct role in dopamine neurotransmission. Synapse 2:125–145, 2000. © 2000 Wiley‐Liss, Inc.

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