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Targeting sst2A Receptor-Expressing Cells in the Rat Hypothalamus through in Vivo Agonist Stimulation: Neuroanatomical Evidence for a Major Role of this Subtype in Mediating Somatostatin Functions

Institut National de la Santé et de la Recherche Médicale, Unité-549, IFR Broca-Sainte Anne, Centre Paul Broca (Z.C., A.S., J.E., P.D.), 75014 Paris, France; and Department of Medical Anatomy (L.H.), Panum Institute, University of Copenhagen, DK-2200 Copenhagen, Denmark

Address all correspondence and requests for reprints to: Pascal Dournaud, Ph.D., Institut National de la Santé et de la Recherche Médicale, Unité-549, IFR Broca-Sainte Anne, Centre Paul Broca, 2ter rue d’Alésia, 75014 Paris, France. E-mail: dournaud{at}broca.inserm.fr.

Numerous physiological studies as well as in situ hybridization and PCR experiments concur in reporting a role for the sst2A receptor in transducing somatostatin (SRIF) actions in the rat hypothalamus. However, the distribution of this receptor protein is not known within this structure. Regional and cellular localization of the sst2A receptor was therefore examined in the rat hypothalamus using highly sensitive immunohistochemical techniques. In close correspondence with the distribution of SRIF-immunoreactive fibers, numerous hypothalamic areas displayed sst2A receptor immunoreactivity. Receptor labeling was, however, diffusely distributed over the tissue, and few immunopositive cells were apparent. Unraveling the distribution of receptor-expressing cells was achieved through acute in vivo agonist stimulation and subsequent receptor internalization. At the cellular level, double-immunolabeling experiments with synaptophysin and microtubule-associated protein 2 demonstrated that sst2A receptors were predominantly internalized in perikarya and dendrites. Double-labeling experiments with SRIF revealed that 93% of arcuate, but only 18% of periventricular, SRIF-positive neurons expressed internalized receptors. Taken together, these results demonstrate for the first time that the sst2A receptor protein is widely, but selectively, distributed in the hypothalamus, and that postsynaptic sst2A auto- and heteroreceptors are well poised to play an important role in the somatostatinergic regulation of hypothalamic endocrine and metabolic processes.

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