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Somatostatin Inhibits Stem Cell Factor Messenger RNA Expression by Sertoli Cells and Stem Cell Factor-Induced DNA Synthesis in Isolated Seminiferous Tubules¹

^a Institut National de la Santé et de la Recherche Médicale (INSERM) Unité 407, Faculté de Médecine Lyon Sud, F-69921 Oullins, France ^b Neurosciences et Systèmes Sensoriels, Université Claude Bernard-Lyon 1, F-69622 Villeurbanne, France ^c Institut National de la Santé et de la Recherche Médicale (INSERM) Unité 531, Institut Fédératif de Recherche 31, Centre Hospitalier Universitaire Rangueil, F-31403 Toulouse, France ^d Laboratoire de Neurosciences, Unité de Formation et de Recherche de Franche-Comté, F-25030 Besançon, France

Immature porcine Sertoli cells have been reported to be targets for the regulatory peptide somatostatin (SRIF), which inhibits the basal and FSH-induced proliferation of Sertoli cells through a decrease of cAMP production. In the present study, we show that SRIF inhibits both basal and FSH-stimulated expression of the stem cell factor (SCF), a Sertoli cell-specific gene. The SRIF-mediated inhibition of forskolin-triggered, but not of 8-bromoadenosine-cAMP-triggered, SCF mRNA expression demonstrates the involvement of adenylyl cyclase in underlying peptide actions. Moreover, these effects require functional coupling of specific plasma membrane receptors to adenylyl cyclase via inhibitory G proteins, because pertussis toxin prevents SRIF-mediated inhibition of SCF mRNA expression. Reverse transcription-polymerase chain reaction (RT-PCR) and Western blot assays suggest the involvement of sst2 receptors in SRIF actions on Sertoli cells. The biological relevance of these data is supported by an SRIF-mediated decrease in SCF-induced incorporation of [³H]thymidine in isolated seminiferous tubules. In situ hybridization and confocal microscopy show that, in seminiferous tubules only, spermatogonia display both c-kit and sst2 receptors. Taken together, these results suggest that SCF-stimulated DNA synthesis can be inhibited by SRIF in spermatogonia, but not in Sertoli and peritubular cells. Combined RT-PCR and immunohistochemical approaches point toward spermatogonia and Leydig cells as the source of testicular SRIF. These data argue in favor of paracrine/autocrine SRIF actions in testis.

First decision: 31 January 2001.

¹ Supported by INSERM (Unité 407), ARC grant 7244 (S.K.) and Ligue contre le Cancer, Comité de la Savoie (S.K.).

² Correspondence and current address: Slavica Krantic, Interactions Cellulaires Neuroendocriniennes, UMR 6544 CNRS-Université de la Méditerranée, Faculté de Médicine Secteur Nord, Boulevard Pierre Dramard, 13916 Marseille, Cedex 20, France. FAX: 33 4 91 69 89 20; krantic.s{at}jean-roche.univ-mrs.fr

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