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The Journal of Neuroscience, August 15, 2000, 20(16):5932-5939

Somatostatin-Induced Regulation of SST_2A Receptor Expression and Cell Surface Availability in Central Neurons: Role of Receptor Internalization

Hélène Boudin¹, Philippe Sarret², Jean Mazella², Agnes Schonbrunn³, and Alain Beaudet¹

¹ Montreal Neurological Institute, McGill University, Montréal, Québec H3A 2B4, Canada, ² Institut de Pharmacologie Moléculaire et Cellulaire, Centre National de la Recherche Scientifique, Université de Nice-Sophia Antipolis, Valbonne, France, and ³ University of Texas, Houston Medical School, Houston, Texas 77225

To investigate the effects of somatostatin (somatotropin release-inhibiting factor, SRIF) on the regulation of SST_2A receptors in mammalian brain, we examined how blockade of SRIF release or stimulation by the SRIF analog [D-Trp⁸]-SRIF would affect the expression and cell surface availability of SST_2A receptors in rat brain slices. First, we measured the intensity of SST_2A immunoreactivity, using quantitative light microscopic immunocytochemistry, and levels of SST_2A mRNA, using semiquantitative RT-PCR, under conditions of acute SRIF release blockade. Incubation of slices from the claustrum or basolateral amygdala, two regions previously shown to contain high concentrations of SST_2A receptors, in Ca²⁺-free Ringer's for 40 min induced a decrease in the intensity of SST_2A receptor immunoreactivity and concentration of SST_2A mRNA as compared with control values obtained in Ca²⁺-supplemented Ringer's. These effects were counteracted in a dose-dependent manner by the addition of 10-100 nM [D-Trp⁸]-SRIF to the Ca²⁺-free medium. Furthermore, both of these effects were abolished in the presence of the endocytosis inhibitors phenylarsine oxide or hyperosmolar sucrose, suggesting that they were dependent on receptor internalization. Electron microscopic immunogold labeling confirmed the existence of an agonist-induced internalization of SST_2A receptors in central neurons. At a high (10 µM), but not at a low (10 nM), concentration of agonist this internalization resulted in a significant decrease in cell surface receptor density, irrespective of the presence of Ca²⁺ in the medium. Taken together, these results suggest that ligand-induced endocytosis is responsible for rapid transcriptional (increase in SST_2A expression) and trafficking (loss of cell surface receptors) events involved in the control of the somatostatinergic signal.

Key words: somatostatin; endocytosis; receptor; immunocytochemistry; electron microscopy; signaling

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Copyright © 2000 Society for Neuroscience  0270-6474/00/20165932-08$05.00/0

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