Messenger ribonucleic acids encoding a serotonin receptor and a novel gene are induced in Sertoli cells by a secreted factor(s) from male rat meiotic germ cells.
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 microM) 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.
Syed
V
V
Center for Research on Reproduction and Women's Health and Department of Obstetrics and Gynecology, University of Pennsylvania, Philadelphia 19104, USA.
Gomez
E
E
Hecht
N B
NB
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D018920:Coculture Techniques; D008297:Male; D008540:Meiosis; D008969:Molecular Sequence Data; D012333:RNA, Messenger / Q000096:biosynthesis*; D051381:Rats; D017207:Rats, Sprague-Dawley; D011985:Receptors, Serotonin / Q000737:chemistry / Q000235:genetics*; D012701:Serotonin / Q000494:pharmacology; D012702:Serotonin Antagonists / Q000494:pharmacology; D017366:Serotonin Receptor Agonists / Q000494:pharmacology; D012708:Sertoli Cells / Q000378:metabolism*; D013094:Spermatozoa / Q000166:cytology / Q000378:metabolism*
D012333:RNA, Messenger; D011985:Receptors, Serotonin; D012702:Serotonin Antagonists; D017366:Serotonin Receptor Agonists; D012701:Serotonin