This Article Full Text Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Request Copyright Permission Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Ge, R.-S. Articles by Hardy, M. P. Search for Related Content PubMed PubMed Citation Articles by Ge, R.-S. Articles by Hardy, M. P. Pubmed/NCBI databases Gene GEO Profiles HomoloGene UniGene Compound via MeSH Substance via MeSH Medline Plus Health Information Steroids Hazardous Substances DB TESTOSTERONE

11ß-Hydroxysteroid Dehydrogenase 2 in Rat Leydig Cells: Its Role in Blunting Glucocorticoid Action at Physiological Levels of Substrate

Population Council (R.-S.G., Q. D., E-m.N., C.M.S., D.O.H., J.F.C., M.P.H.) and The Rockefeller University (M.P.H., J.F.C.), New York, New York 10021; and Department of Pathology and Laboratory Medicine (S.A.L., D.J.M.), The Miriam Hospital, Brown University School of Medicine, Providence, Rhode Island 02906

Address all correspondence and requests for reprints to: Matthew P. Hardy, The Population Council, 1230 York Avenue, New York, New York 10021. E-mail: m-hardy{at}popcbr.rockefeller.edu.

Corticosterone (CORT) suppresses Leydig cell steroidogenesis by inhibiting the expression of proteins involved in testosterone biosynthesis including steroidogenic acute regulatory protein and steroidogenic enzymes. In most cells, intracellular glucocorticoid levels are controlled by either or both of the two known isoforms of 11ß-hydroxysteroid dehydrogenase (11ß HSD): the nicotinamide adenine dinucleotide phosphate reduced-dependent low-affinity type I 11ß HSD (11ß HSD1) oxidoreductase and the nicotinamide adenine dinucleotide-dependent 11ß HSD2 high-affinity unidirectional oxidase. In Leydig cells, 11ß HSD1 alone may not be sufficient to prevent glucocorticoid-mediated suppression due to its low affinity for CORT at basal concentrations. The high-affinity unidirectional 11ß HSD2, if also present, may be critical for lowering intracellular CORT levels. In the present study, we showed that 11ß HSD2 is present in rat Leydig cells by PCR amplification, immunohistochemical staining, enzyme histochemistry, immunoprecipitation, and Western blotting. Real-time PCR showed a 6-fold enrichment of 11ß HSD2 mRNA in these cells, compared with whole testis and that the amount of 11ß HSD2 message was about 1000-fold lower, compared with 11ß HSD1. Diffuse immunofluorescent staining of 11ß HSD2 protein in the Leydig cell cytoplasm was consistent with its localization in the smooth endoplasm reticulum. 11ß HSD1 or 11ß HSD2 activities were selectively inhibited using antisense methodology: inhibition of 11ß HSD1 lowered reductase activity by 60% and oxidation by 25%, whereas inhibition of 11ß HSD2 alone suppressed oxidase activity by 50%. This shows that the high-affinity, low-capacity 11ß HSD2 isoform, present at only one thousandth the level of the low-affinity isoform may significantly affect the level of CORT. The inhibition of either 11ß HSD1 or 11ß HSD2 significantly lowered testosterone production in the presence of CORT. These data suggest that both types I and II 11ß HSD in Leydig cells play a protective role, opposing the adverse effects of excessive CORT on testosterone production.

This article has been cited by other articles:

				G.-X. Hu, H. Lin, C. M. Sottas, D. J. Morris, M. P. Hardy, and R.-S. Ge Inhibition of 11{beta}-Hydroxysteroid Dehydrogenase Enzymatic Activities by Glycyrrhetinic Acid In Vivo Supports Direct Glucocorticoid-Mediated Suppression of Steroidogenesis in Leydig Cells J Androl, May 1, 2008; 29(3): 345 - 351. [Abstract] [Full Text] [PDF]

				C Chandras, T E Harris, A L. Bernal, D R E Abayasekara, and A E Michael PTGER1 and PTGER2 receptors mediate regulation of progesterone synthesis and type 1 11{beta}-hydroxysteroid dehydrogenase activity by prostaglandin E2 in human granulosa lutein cells J. Endocrinol., September 1, 2007; 194(3): 595 - 602. [Abstract] [Full Text] [PDF]

				V. Sharp, L. M Thurston, R. C Fowkes, and A. E Michael 11{beta}-Hydroxysteroid dehydrogenase enzymes in the testis and male reproductive tract of the boar (Sus scrofa domestica) indicate local roles for glucocorticoids in male reproductive physiology Reproduction, September 1, 2007; 134(3): 473 - 482. [Abstract] [Full Text] [PDF]

				N. Sunak, D. F Green, L. R Abeydeera, L. M Thurston, and A. E Michael Implication of cortisol and 11{beta}-hydroxysteroid dehydrogenase enzymes in the development of porcine (Sus scrofa domestica) ovarian follicles and cysts Reproduction, June 1, 2007; 133(6): 1149 - 1158. [Abstract] [Full Text] [PDF]

				L M Thurston, D R E Abayasekara, and A E Michael 11{beta}-Hydroxysteroid dehydrogenase expression and activities in bovine granulosa cells and corpora lutea implicate corticosteroids in bovine ovarian physiology J. Endocrinol., May 1, 2007; 193(2): 299 - 310. [Abstract] [Full Text] [PDF]

				K. C. Jonas, C. Chandras, D. R. E. Abayasekara, and A. E. Michael Role for Prostaglandins in the Regulation of Type 1 11{beta}-Hydroxysteroid Dehydrogenase in Human Granulosa-Lutein Cells Endocrinology, December 1, 2006; 147(12): 5865 - 5872. [Abstract] [Full Text] [PDF]

				Y. Ozaki, M. Higuchi, C. Miura, S. Yamaguchi, Y. Tozawa, and T. Miura Roles of 11{beta}-Hydroxysteroid Dehydrogenase in Fish Spermatogenesis Endocrinology, November 1, 2006; 147(11): 5139 - 5146. [Abstract] [Full Text] [PDF]

				T S. Kavitha, C Parthasarathy, R Sivakumar, R Badrinarayanan, and K Balasubramanian Effects of excess corticosterone on NADPH generating enzymes and glucose oxidation in Leydig cells of adult rats Human and Experimental Toxicology, March 1, 2006; 25(3): 119 - 125. [Abstract] [PDF]

				S. K. Paulsen, S. B. Pedersen, J. O. L. Jorgensen, S. Fisker, J. S. Christiansen, A. Flyvbjerg, and B. Richelsen Growth Hormone (GH) Substitution in GH-Deficient Patients Inhibits 11{beta}-Hydroxysteroid Dehydrogenase Type 1 Messenger Ribonucleic Acid Expression in Adipose Tissue J. Clin. Endocrinol. Metab., March 1, 2006; 91(3): 1093 - 1098. [Abstract] [Full Text] [PDF]

				G. Shetty, C. C. Y. Weng, S. J. Meachem, O. U. Bolden-Tiller, Z. Zhang, P. Pakarinen, I. Huhtaniemi, and M. L. Meistrich Both Testosterone and Follicle-Stimulating Hormone Independently Inhibit Spermatogonial Differentiation in Irradiated Rats Endocrinology, January 1, 2006; 147(1): 472 - 482. [Abstract] [Full Text] [PDF]