Critical role of nitric oxide in expression of porcine LH receptor at transcription and post-transcription levels

Summary:

The present study was performed to clarify the involvement of nitric oxide (NO) in the expression of LH receptor in porcine granulosa cells. The granulosa cells, prepared from porcine ovarian follicles, were developed in the presence of FSH. LH receptor mRNA was induced to reach a maximal level after a 24-h culture with FSH, as determined by semi-quantitative reverse transcriptase-PCR (RT-PCR). In our previous report ([Nishida et al., 2000]), we found that NO was released from granulosa cells after a 40-48 h culture with FSH. When 200 μM NO scavenger was added to cultures before the NO release (30 h), the content of LH receptor on the cells decreased to 28% that of the control. However, the receptor content was not influenced by addition of NO scavenger at 46 h, or by 50 μM NO donor at 30 or 46 h. During transformation of mature granulosa cells to luteal cells, LH receptor mRNA was induced after a 24-h culture with LH, which induction was inhibited by removal of endogenous NO. However, the expression was not influenced by addition of either NO scavenger at 46 h or by NO donor. During the period of these treatments, cellular DNA contents were constant. Consequently, the transient generation of NO may play a critical role in expression of the LH receptor at transcription and post-transcription levels.

Key words:

Nitric oxide - LH receptor - granulosa cells

References

• 1 Ben-Shlomo I, Kokia E, Jackson M J, Adashi E Y, Payne D W. Interleukin-1β stimulates nitrite production in the rat ovary: evidence for heterologous cell-cell interaction and for insulin-mediated regulation of the inducible isoform of nitric oxide synthase.  Biol Reprod. 51 310-318 1994; 
• 2 Bonello N, Mckie K, Jasper M, Andrew L, Ross N, Braybon E, Brannstrom M, Norman R J. Inhibition of nitric oxide: effects on interleukin-1β-enhanced ovulation rate, steroid hormone, and ovarian leukocyte distribution at ovulation in the rat.  Biol Reprod. 54 436-445 1996; 
• 3 Bredt D S, Hwang P M, Glatt C E, Lowenstein C, Reed R R, Snyder S H. Cloned and expressed nitric oxide synthase structurally resembles cytochrome P-450 reductase.  Nature Lond. 351 714-718 1991; 
• 4 Bredt D S, Snyder S H. Nitric oxide mediates glutamate-linked enhancement of cGMP levels in the cerebellum.  Proc Natl Acad Sci USA. 86 9030-9033 1989; 
• 5 Chun S-Y, Eisenhauer K M, Kubo M, Hsueh A JW. Interleukin-1β suppresses apoptosis in rat ovarian follicles by increasing nitric oxide production.  Endocrinology. 136 3120-3127 1995; 
• 6 Clementi E. Role of nitric oxide and its intracellular signalling pathways in the control of Ca²⁺ homeostasis.  Biochem Pharmacol. 55 713-718 1998; 
• 7 Doucet J P, Squinto S P, Bazan N G. Fos-jun and the primary genomic response in the nervous system. Possible physiological role and pathophysiological significance.  Mol Neurobiol. 4 27-55 1990; 
• 8 Dufau M L, Winters C A, Hattori M-A, Aquilano D R, Baranao J LS, Nozu K, Baukal A, Catt K J. Homronal regulation of androgen production by the Leydig cell.  J Steroid Biochem. 20 161-173 1984; 
• 9 El-Hefnawy T, Krawczyk Z, Nikula H, Vihera I, Huhtaniemi I. Regulation of function of the murine luteinizing hormone receptor promoter by cis- and trans-acting elements in mouse Leydig tumor cells.  Mol Cell Endocrinol. 119 207-217 1996; 
• 10 Ellman C, Corbett J A, Misko T P, McDaniel M, Beckerman K P. Nitric oxide mediates interleukin-1-induced cellular cytotoxicity in the rat ovary: a potential role for nitric oxide in the ovulatory process.  J Clin Invest. 92 3053-3056 1993; 
• 11 Hattori M-A, Nishida N, Takesue K, Kato Y, Fujihara N. Nitric oxide synthesis by porcine oocytes: its regulation and function during follicular development.  Biol Reprod 60 ((Suppl. 1)) A445 1999; 
• 12 Hattori M-A, Sakamoto K, Fujihara N, Kojima I. Nitric oxide: a modulator for the epidermal growth factor receptor expression in developing ovarian granulosa cells.  Am J Physiol. 270 C812-C818 1996; 
• 13 Hattori M-A, Hachisu T, Shimohigashi Y, Wakabayashi K. Conformation of the β subunit of deglycosylated human chorionic gonadotropin in the interaction at receptor sites.  Mol Cell Endocrinol. 57 17-23 1988; 
• 14 Hattori M-A, Ozawa K, Wakabayashi K. Sialic acid is responsible for the charge heterogeneity and the biological potency of rat lutropin.  Biochem Biophys Res Commun. 127 501-508 1985; 
• 15 Hsueh A JW, Bicsak T A, Jia X-C, Dahl K D, Fauser B CJM, Galway A B, Czekala N, Pavlou S N, Papkoff H, Keene J, Boime I. Granulosa cells as hormone target: the role of biologically active follicle-stimulating hormone in reproduction.  Recent Prog Horm Res. 45 209-273 1989; 
• 16 Hu Y L, Lei Z M, Rao C V. Cis-acting elements and trans-acting proteins in the transcription of chorionic gonadotropin/luteinizing hormone receptor gene in human choriocarcinoma cells and placenta.  Endocrinology. 137 3897-3905 1996; 
• 17 Jablonka-Shariff A, Olson L M. Hormonal regulation of nitric oxide synthases and their cell-specific expression during follicular development in the rat ovary.  Endocrinology. 138 460-468 1997; 
• 18 Kwon N S, Nathan C F, Gilker C, Griffith O W, Matthews D E, Stuehr D J. l-Citrulline production from l-arginine by macrophage nitric oxide synthase.  J Biol Chem. 265 13442-13445 1990; 
• 19 Lamas S, Marsden P A, Li G K, Tempst P, Michel T. Endothelial nitric oxide synthase: molecular cloning and characterization of a distinct constitutive enzyme isoform.  Proc Natl Acad Sci USA. 89 6348-6352 1992; 
• 20 Loosfelt H, Misrahi M, Atger M, Salesse R, Vu Hai-Luu Thi M T, Jolivet A, Guiochon-Mantel A, Sar S, Jallal B, Garnier J, Milgrom E. Cloning and sequencing of porcine LH-hCG receptor cDNA: Variants lacking transmembrane domain.  Science. 245 525-528 1989; 
• 21 Lowenstein C J, Glatt C S, Bredt D S, Snyder S H. Cloned and expressed macrophage nitric oxide synthase contrasts with the brain enzyme.  Proc Natl Acad Sci USA. 89 6711-6715 1992; 
• 22 Mondschein J S, Smith S A, Hammond J M. Production of insulin-like growth factor binding proteins (IGFBPs) by porcine granulosa cells: identification of IGFBP-2 and -3 and regulation by hormones and growth factors.  Endocrinology. 127 2298-2306 1990; 
• 23 Nishida N, Takesue K, Hattori M-A, Kato Y, Wakabayashi K, Fujihara N. Modulatory action of nitric oxide on the expression of transcription factor genes, c-fos and c-jun, in developing porcine granulosa cells.  J Reprod Develop. 46 167-175 2000; 
• 24 Palmer R MJ, Moncada S. A novel citrulline-forming enzyme implicated in the formation of nitric oxide by vascular endothelial cells.  Biochem Biophys Res Commun. 158 348-353 1989; 
• 25 Richards J S, Fitzpatrick S L, Clemens J W, Morris J K, Alliston T, Sirois J. Ovarian cell differentiation: a cascade of multiple hormones, cellular signals, and regulated genes.  Recent Prog Horm Res. 50 223-254 1995; 
• 26 Sheng M, Greenberg M E. The regulation and function of c-fos and other immediate early genes in the nervous system.  Neuron. 4 477-485 1990; 
• 27 Shukovski L, Tsafriri T. The involvement of nitric oxide in the ovulatory process in the rat.  Endocrinology. 135 2287-2290 1995; 
• 28 Singh S, Siminovitch D. A reliable method for the estimation of DNA in higher plant tissues.  Anal Biochem. 711 308-312 1976; 
• 29 Stamler J S, Singel D J, Loscalzo J. Biochemistry of nitric oxide and its redox-activated forms.  Science. 258 1898-1902 1992; 
• 30 Tabraue C, Penate R D, Gallardo G, Hernandez I, Quintana J, Blanco F L, Reyes J G, Fanjul L F, de Galarreta C MR. Induction of guanosine triphosphate-cyclohydrolase by follicle-stimulating hormone enhances interleukin-1β-stimulated nitric oxide synthase activity in granulosa cells.  Endocrinology. 138 162-168 1997; 
• 31 Yamauchi J, Miyazaki T, Iwasaki S, Kishi I, Kuroshima M, Tei C, Yoshimura Y. Effects of nitric oxide on ovulation and ovarian steroidogenesis and prostaglandin production in the rabbit.  Endocrinology. 138 3630-3637 1997;