Abstract
P2Y receptors are G protein-coupled receptors composed of eight known subunits (P2Y(1, 2, 4, 6, 11, 12, 13, 14)), which are involved in different functions in neural tissue. The present study investigates the expression pattern of P2Y(4) receptors in the rat central nervous system (CNS) using immunohistochemistry and in situ hybridization. The specificity of the immunostaining has been verified by preabsorption, Western blot, and combined use of immunohistochemistry and in situ hybridization. Neurons expressing P2Y(4) receptors were distributed widely in the rat CNS. Heavy P2Y(4) receptor immunostaining was observed in the magnocellular neuroendocrine neurons of the hypothalamus, red nucleus, pontine nuclei, mesencephalic trigeminal nucleus, motor trigeminal nucleus, ambiguous nucleus, inferior olive, hypoglossal nucleus, and dorsal motor vagus nucleus. Both neurons and astrocytes express P2Y(4) receptors. P2Y(4) receptor immunostaining signals were mainly confined to cell bodies and dendrites of neurons, suggesting that P2Y(4) receptors are mainly involved in regulating postsynaptic events. In the hypothalamus, all the vasopressin (VP) and oxytocin (OT) neurons and all the orexin A neurons were immunoreactive for P2Y(4) receptors. All the neurons expressing P2Y(4) receptors were found to express N-methyl-D: -aspartate receptor 1 (NR1). These data suggest that purines and pyrimidines might be involved in regulation of the release of the neuropeptides VP, OT, and orexin in the rat hypothalamus via P2Y(4) receptors. Further, the physiological and pathophysiological functions of the neurons may operate through coupling between P2Y(4) receptors and NR1.