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Center for Research on Reproduction and Women’s Health and Department of Obstetrics and Gynecology, University of Pennsylvania (V.S., N.B.H.), Philadelphia, Pennsylvania 19104
Address all correspondence and requests for reprints to: Dr. Norman B. Hecht, Center for Research on Reproduction and Women’s Health, 421 Curie Boulevard, 1310 BRB II-III/6142, University of Pennsylvania, Philadelphia, Pennsylvania 19104. E-mail: nhecht{at}mail.med.upenn.edu
Using Sertoli cell-germ cell cocultures and messenger RNA (mRNA) differential display, we have identified a complementary DNA of 355 nucleotides that is up-regulated in Sertoli cells by pachytene spermatocytes. The mRNA differential display pattern was confirmed by Northern blotting. Sequence analysis revealed a homology of 91% (nt) and 86% (aa) to a serotonin receptor. The mRNA encoding the serotonin receptor was detected in Sertoli cells after 18 h of coculture. Its induction did not require cell contact, as germ cell-conditioned medium also induced the mRNA. The germ cell factor(s) inducing the serotonin receptor mRNA is more than 10 kDa, survives freezing and thawing, and is heat sensitive. A high dose of serotonin (10 µM) or the serotonin receptor agonists (±)-1-(2,5-dimethoxy-4-iodophenyl)-2-aminopropane HCl and quipazine induce the serotonin receptor mRNA in Sertoli cells after 24 h. The antagonists, ketanserin and spiperone, inhibit the serotonin-mediated mRNA induction but fail to inhibit the germ cell-mediated induction, suggesting that the germ cell factor(s) up-regulates the serotonin receptor by a distinct pathway. A second clone of 380 nucleotides, induced in Sertoli cells by pachytene spermatocytes or germ cell-conditioned medium, did not show homology to database sequences. The germ cell factor(s) inducing the second clone is larger than 10 kDa, but is inactivated by freezing/thawing and boiling. The induction of a serotonin receptor mRNA and a second novel mRNA in Sertoli cells by pachytene spermatocytes demonstrates that meiotic germ cells induce mRNA encoding an important receptor in Sertoli cells.
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