title: Nitric oxide: An autocrine regulator of Leydig cell steroidogenesis in the Asian catfish, Clarias batrachus.
abstract: Nitric oxide has been recognized as an important inter- and intra-cellular modulator of testicular steroidogenesis in higher vertebrates with conflicting results. Moreover, its role in regulation of testicular steroidogenesis in ectothermic vertebrates is non-existent. The present study was, therefore, undertaken to examine whether Leydig cells of a freshwater catfish, Clarias batrachus produce nitric oxide (NO), if so, does it regulate its steroidogenic activity? The purified Leydig cells were stained histochemically for NADPH-diaphorase (NADPH-d) activity, and immunocytochemically for neuronal and inducible nitric oxide synthase (nNOS and iNOS) like molecules. Leydig cells were also incubated with NOS inhibitor, N-nitro-l-arginine methyl ester (l-NAME), and NO donor, sodium nitroprusside (SNP). NO and testosterone released in incubation medium were analyzed. A distinct positive NAPDH-d staining was observed in Leydig cells. These cells also exhibited immunoprecipitation of variable intensity with nNOS and iNOS antibodies. Further, l-NAME treatment caused significant suppression in NO production and elevation in testosterone secretion by Leydig cells. On the contrary, exposure of Leydig cells to SNP resulted in increased NO production with concomitant decline in testosterone level. Thus, the present study reports NO production by Leydig cells in fish for the first time, which appears to inhibit its own androgen production.
authors.affiliation: Fish Endocrinology Laboratory, Department of Zoology, Banaras Hindu University, Varanasi 221 005, UP, India.
authors.forename: Neelima Dubey
authors.initials: ND
authors.lastname: nee Pathak
authors.forename: Bechan
authors.initials: B
authors.lastname: Lal
mesh_terms: D000818:Animals; D002397:Catfishes / Q000378:metabolism*; D004791:Enzyme Inhibitors / Q000494:pharmacology; D007150:Immunohistochemistry / Q000662:veterinary; D007985:Leydig Cells / Q000187:drug effects / Q000201:enzymology / Q000378:metabolism; D008297:Male; D008857:Microscopy, Interference / Q000662:veterinary; D009252:NADPH Dehydrogenase / Q000378:metabolism; D019331:NG-Nitroarginine Methyl Ester / Q000494:pharmacology; D009569:Nitric Oxide / Q000378:metabolism*; D020030:Nitric Oxide Donors / Q000494:pharmacology; D052248:Nitric Oxide Synthase Type I / Q000378:metabolism*; D052247:Nitric Oxide Synthase Type II / Q000378:metabolism*; D009599:Nitroprusside / Q000494:pharmacology; D013737:Testis / Q000187:drug effects / Q000201:enzymology / Q000378:metabolism*; D013739:Testosterone / Q000096:biosynthesis*
chemical_list: D004791:Enzyme Inhibitors; D020030:Nitric Oxide Donors; D009599:Nitroprusside; D009569:Nitric Oxide; D013739:Testosterone; D052248:Nitric Oxide Synthase Type I; D052247:Nitric Oxide Synthase Type II; D009252:NADPH Dehydrogenase; D019331:NG-Nitroarginine Methyl Ester