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The Journal of Neuroscience, December 1, 2000, 20(23):8677-8684

Dopamine D₃ Receptors Expressed by All Mesencephalic Dopamine Neurons

Jorge Diaz¹, Catherine Pilon², Bernard Le Foll², Claude Gros², Antoine Triller³, Jean-Charles Schwartz², and Pierre Sokoloff²

¹ Laboratoire de Physiologie, Université René Descartes, 75006 Paris, France, ² Unité de Neurobiologie et Pharmacologie Moléculaire (Institut National de la Santé et de la Recherche Médicale U 109), Centre Paul Broca, 75014 Paris, France, and ³ Biologie Cellulaire de la Synapse Normale et Pathologique (Institut National de la Santé et de la Recherche Médicale U 497), Ecole Normale Supérieure, 75230 Paris, France

A polyclonal antibody was generated using synthetic peptides designed in a specific sequence of the rat D₃ receptor (D₃R). Using transfected cells expressing recombinant D₃R, but not D₂ receptor, this antibody labeled 45-80 kDa species in Western blot analysis, immunoprecipitated a soluble fraction of [¹²⁵I]iodosulpride binding, and generated immunofluorescence, mainly in the cytoplasmic perinuclear region of the cells. In rat brain, the distribution of immunoreactivity matched that of D₃R binding, revealed using [¹²⁵I]R(+)trans-7-hydroxy-2-[N-propyl-N-(3'-iodo-2'-propenyl)amino] tetralin ([¹²⁵I]7-trans-OH-PIPAT), with dense signals in the islands of Calleja and mammillary bodies, and moderate to low signals in the shell of nucleus accumbens (AccSh), frontoparietal cortex, substantia nigra (SN), ventral tegmental area (VTA) and lobules 9 and 10 of the cerebellum. Very low or no signals could be detected in other rat brain regions, including dorsal striatum, or in D₃R-deficient mouse brain. Labeling of perikarya of AccSh and SN/VTA appeared with a characteristic punctuate distribution, mostly at the plasma membrane where it was not associated with synaptic boutons, as revealed by synaptophysin immunoreactivity. In SN/VTA, D₃R immunoreactivity was found on afferent terminals, arising from AccSh, in which destruction of intrinsic neurons by kainate infusions produced a loss of D₃R binding in both AccSh and SN/VTA. D₃R-immunoreactivity was also found in all tyrosine hydroxylase (TH)-positive neurons observed in SN, VTA and A8 retrorubral fields, where it could represent D₃ autoreceptors controlling dopamine neuron activities, in agreement with the elevated dopamine extracellular levels in projection areas of these neurons found in D₃R-deficient mice.

Key words: nucleus accumbens shell; substantia nigra; ventral tegmental area; D₃ receptor-deficient mice; tyrosine hydroxylase; synaptophysin

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Copyright © 2000 Society for Neuroscience  0270-6474/00/20238677-08$05.00/0

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