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Endocrinology Vol. 139, No. 8 3637-3645
Copyright © 1998 by The Endocrine Society

ARTICLES

Expression and Regulation of Interferon-{gamma}-Inducible Protein 10 Gene in Rat Leydig Cells1

Jianxin Hu, Shaojin You, Wei Li, Deli Wang, Madan L. Nagpal, Yide Mi, Peng Liang and Tu Lin

Research and Medical Service, WJB Dorn Veterans Medical Center, and the Department of Medicine, University of South Carolina School of Medicine (J.H., S.Y., W.L., D.W., M.L.N., Y.M., T.L.), Columbia, South Carolina 29208; and the Vanderbilt Cancer Center, Department of Cell Biology, Vanderbilt University School of Medicine (P.L.), Nashville, Tennessee 37232

Address all correspondence and requests for reprints to: Tu Lin, M.D., Department of Medicine, University of South Carolina School of Medicine, Medical Library Building, Suite 316, Columbia, South Carolina 29208.

In the present study, we report the cloning of a gene that is differentially expressed in normal adult rat Leydig cells and whose expression is inhibited by hCG but is induced by interferon-{gamma} (IFN{gamma}). DNA sequence analysis identified this gene as rat IFN{gamma}-inducible protein 10 (IP-10), a member of the -C-X-C- chemokine superfamily of proinflammatory cytokines. High levels of IP-10 messenger RNA (mRNA) were constitutively expressed in freshly isolated and primary cultured Leydig cells. hCG inhibited this expression in a dose-dependent manner. The addition of 1 ng/ml hCG inhibited IP-10 mRNA levels more than 80%. Conversely, IP-10 mRNA levels were markedly increased in response to murine interleukin-1{alpha}, murine tumor necrosis factor-{alpha}, and murine IFN{gamma} by 3.3-, 10-, and 26-fold, respectively. Concomitant addition of murine interleukin-1{alpha}, murine tumor necrosis factor-{alpha}, and murine IFN{gamma} synergistically increased IP-10 mRNA levels by 58-fold. Furthermore, in addition to one previously described rat IP-10 mRNA transcript (1.5 kb), another larger transcript (2.7 kb) was identified by Northern blot in rat Leydig cells. After screening a rat testis complementary DNA library, we obtained a partial structural gene and an intron sequence, which possibly originated from the larger transcript of rat IP-10 mRNA. Histochemical and immunocytochemical staining revealed that purified cells were positive for 3ß-hydroxysteroid dehydrogenase and IP-10, confirming that IP-10 is indeed present in Leydig cells. IP-10 antisense oligonucleotides enhanced basal and hCG-induced testosterone formation. This suggests that endogenous IP-10 has an inhibitory effect on Leydig cell steroidogenesis. In conclusion, IP-10 is expressed in rat Leydig cells and may have paracrine and autocrine effects on testicular function.




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