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Laboratoire de Neurosciences et Systèmes Sensoriels (A.B.H., K.A.J., P.D.-V.), Centre National de la Recherche Scientifique, Unité Mixte de Recherche 5020, Université Claude Bernard, 69366 Lyon cedex 07; and Laboratoire Neurobiologie de l’Olfaction et Prise Alimentaire (J.A., C.B., M.C., R.S.), Equipe Récepteurs et Communication Chimique, Institut National de la Recherche Agronomique, 78352 Jouy-en-Josas, France
Address all correspondence and requests for reprints to: Patricia Duchamp-Viret, Laboratoire de Neurosciences et Systèmes Sensoriels, Centre National de la Recherche Scientifique, Unité Mixte de Recherche 5020, Université Claude Bernard, 50 Avenue Tony Garnier, 69366 Lyon cedex 07, France. E-mail: pduchamp{at}olfac.univ-lyon1.fr.
Orexin A and B are involved in feeding behaviors, and recently fibers containing these peptides were found in the rat olfactory bulb. These fibers, which originate from the lateral and posterior hypothalamus and the perifornical area, are distributed in the glomerular, mitral cell, and granule cell layers. Orexin receptors are mainly expressed by mitral cells. In the present study, RT-PCR experiments were done to determine orexin receptor expression during the early postnatal life of rats, and immunocytochemical experiments were performed to further clarify the structural and ultrastructural localization of orexin receptors in the olfactory bulb. Furthermore, a functional electrophysiological approach examined the action of orexin A on mitral cell excitability and spontaneous activity using in vitro patch-clamp techniques. RT-PCR results show that mRNA of the two type receptors, type 1 orexin receptors and type 2 orexin receptors, are expressed in the olfactory bulb of rat from 10 d to the adult stage. At the same ages, immunocytochemical data show that orexin 1 receptors are localized in the cell bodies of periglomerular, mitral/tufted, and granule cells. Immunoreactivity was also demonstrated in mitral/tufted cell dendrites arborizing in the glomerulus and mitral/tufted and granule cell processes running in the external plexiform layer. Functionally, orexin A produced either a direct, tetrodotoxin-insensitive depolarization in one group of mitral cells (7%), or, in another group (30%), an indirect, tetrodotoxin-sensitive hyperpolarization. Both actions were mediated by type 1 orexin receptors because the response was antagonized by SB-334867-A, a selective antagonist. Mitral cell recordings performed under bicuculline [
-aminobutyric acid (GABA)A receptor antagonist], indicate that the orexin-induced indirect hyperpolarization was partly mediated through GABAA receptors. Because granule cells and periglomerular cells express orexin receptors and are GABAergic cells, they could be both involved in this hyperpolarization. Other mechanisms, which could support an indirect hyperpolarization of mitral cells through dopamine interneuron solicitation, are proposed. Our results provide data that should allow us to better understand neural communication and regulation mechanisms between the hypothalamic feeding centers and the olfactory bulb.
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