This Article Full Text Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Weissman, B. A. Articles by Hardy, M. P. Search for Related Content PubMed PubMed Citation Articles by Weissman, B. A. Articles by Hardy, M. P.

Paracrine Modulation of Androgen Synthesis in Rat Leydig Cells by Nitric Oxide

ENMEI NIU

RENSHAN GE

CHANTAL M. SOTTAS

MICHAEL HOLMES

JAMES C. HUTSON

MATTHEW P. HARDY

From the ^* Department of Pharmacology, Israel Institute for Biological Research, Ness Ziona, Israel; the Center for Biomedical Research, Population Council and the Rockefeller University, New York, New York; and the Department of Cell Biology and Biochemistry, Texas Tech University Health Sciences Center, Lubbock, Texas.

Correspondence to: Dr B. A. Weissman, Department of Pharmacology, Israel Institute for Biological Research, PO Box 19, Ness Ziona 74100, Israel (e-mail: baw{at}iibr.gov.il).

The free radical nitric oxide (NO), generated through the oxidation of L-arginine to L-citrulline by NO synthases (NOSs), has been shown to inhibit steroidogenic pathways. NOS isoforms are known to be present in rat and human testes. Our study examined the sensitivity of Leydig cells to NO and determined whether NOS activity resides in Leydig cells or in another cell type such as the testicular macrophage. The results showed a low level of L-[¹⁴C]arginine conversion in purified rat Leydig cell homogenates. Administration of the NOS inhibitor L-N^G-nitro-arginine methyl ester (L-NAME), or the calcium chelator ethylenebis (oxyethylenenitrilo)tetraacetic acid (EGTA), had no effect on L-[¹⁴C]citrulline accumulation. Increased intracellular Ca²⁺ concentrations that were induced by a calcium ionophore, or the addition of luteinizing hormone (LH), failed to affect NO formation in intact cells that were cultured in vitro. Introduction of a high concentration of the NO precursor L-arginine did not decrease testosterone (T) production, and NOS inhibitors did not increase T biosynthesis. However, exposing Leydig cells to low concentrations of the NO donor S-nitrosoglutathione (GSNO) induced a dramatic blockade of T production under basal and LH-stimulated conditions. DNA array assays showed a low level of expression of endothelial NOS (eNOS), while the neuronal and inducible isoforms of NOS (nNOS and iNOS) were below detection levels. Reverse transcriptase-polymerase chain reaction (RT-PCR) analyses confirmed these findings and demonstrated the presence of high iNOS messenger RNA (mRNA) levels in activated testicular macrophages that produced large amounts of NO. These data suggest that, while T production in rat Leydig cells is highly sensitive to NO and an endogenous NO-generating system is not present in these cells, NOS activity is more likely to reside in activated testicular macrophages.

     Key words: Macrophage, testosterone, DNA gene array, nitric oxide synthase, S-nitrosoglutathione, L-N^G-nitro-arginine methyl ester, L-N^G-monomethyl-arginine

This article has been cited by other articles:

				J. Guo, Y. Jia, S.-X. Tao, Y.-C. Li, X.-S. Zhang, Z.-Y. Hu, N. Chiang, Y.-H. Lue, A. P. S. Hikim, R. S. Swerdloff, et al. Expression of Nitric Oxide Synthase During Germ Cell Apoptosis in Testis of Cynomolgus Monkey After Testosterone and Heat Treatment J Androl, March 1, 2009; 30(2): 190 - 199. [Abstract] [Full Text] [PDF]

				C. Mondillo, R. M. Pagotto, B. Piotrkowski, C. G. Reche, Z. J. Patrignani, C. B. Cymeryng, and O. P. Pignataro Involvement of Nitric Oxide Synthase in the Mechanism of Histamine-Induced Inhibition of Leydig Cell Steroidogenesis via Histamine Receptor Subtypes in Sprague-Dawley Rats Biol Reprod, January 1, 2009; 80(1): 144 - 152. [Abstract] [Full Text] [PDF]

				B. A. Weissman, C. M. Sottas, P. Zhou, C. Iadecola, and M. P. Hardy Testosterone production in mice lacking inducible nitric oxide synthase expression is sensitive to restraint stress Am J Physiol Endocrinol Metab, February 1, 2007; 292(2): E615 - E620. [Abstract] [Full Text] [PDF]

				J. C. Hutson Physiologic Interactions Between Macrophages and Leydig Cells Experimental Biology and Medicine, January 1, 2006; 231(1): 1 - 7. [Abstract] [Full Text] [PDF]

				M. Herman and C. Rivier Activation of a Neural Brain-Testicular Pathway Rapidly Lowers Leydig Cell Levels of the Steroidogenic Acute Regulatory Protein and the Peripheral-Type Benzodiazepine Receptor while Increasing Levels of Neuronal Nitric Oxide Synthase Endocrinology, January 1, 2006; 147(1): 624 - 633. [Abstract] [Full Text] [PDF]