Fertility and Sterility

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Fertility and Sterility Effects of nitric oxide and cGMP on inhibin A and inhibin subunit mRNA levels from cultured rat granulosa cells You-Hsin Jean Chen, M.S., Michael Tafoya, B.S., Anna Ngo, Philip S LaPolt, Ph.D. Fertility and Sterility Volume 79, Pages 687-693 (March 2003) DOI: 10.1016/S0015-0282(02)04823-9

FIGURE 1 Modulation of FSH-stimulated inhibin A levels by DETA/NO and dibutyryl cGMP. Granulosa cells were treated for 48 hours with media alone (C; control), FSH (2 ng/mL), FSH plus DETA/NO (F+D; 100 μM), or FSH plus dibutyryl cGMP (F+cGMP; 2 mM), followed by analysis of inhibin A levels by ELISA. Chen. Effects of NO and cGMP on inhibin A. Fertil Steril 2003. Fertility and Sterility 2003 79, 687-693DOI: (10.1016/S0015-0282(02)04823-9)

FIGURE 2 Influence of DETA/NO and dibutyryl cGMP on FSH-stimulated E2 production. Conditioned media obtained from cells treated in Figure 1 were also subjected to E2 RIA to confirm the inhibitory effect of NO and cGMP treatments on E2 production. Chen. Effects of NO and cGMP on inhibin A. Fertil Steril 2003. Fertility and Sterility 2003 79, 687-693DOI: (10.1016/S0015-0282(02)04823-9)

FIGURE 3 Influence of DETA/NO and dibutyryl cGMP on forskolin-stimulated inhibin A concentrations from rat granulosa cells. Cells were treated for 48 hours with media alone (C), forskolin (FSK; 10−4 M), FSK plus DETA/NO (FSK+D), or FSK plus a cGMP analog (FSK+cGMP). Chen. Effects of NO and cGMP on inhibin A. Fertil Steril 2003. Fertility and Sterility 2003 79, 687-693DOI: (10.1016/S0015-0282(02)04823-9)

FIGURE 4 Inhibition of cGMP production by a soluble guanylyl cyclase inhibitor. Cells were treated for 48 hours with media alone (C), FSH, FSH plus the soluble guanylyl cyclase inhibitor ODQ (FSH+O), FSH plus DETA/NO (FSH+D), or FSH plus DETA/NO plus ODQ (FSH+D+O). After culture, levels of cGMP accumulation were determined by cGMP RIA. Chen. Effects of NO and cGMP on inhibin A. Fertil Steril 2003. Fertility and Sterility 2003 79, 687-693DOI: (10.1016/S0015-0282(02)04823-9)

FIGURE 5 Cyclic GMP–independent effects of NO on inhibin A levels. To determine whether the effects of NO on inhibin A were dependent upon increased cGMP production, cells were treated for 48 hours with media alone (C), FSH, FSH plus the soluble guanylyl cyclase inhibitor ODQ (FSH+O), FSH plus DETA/NO (FSH+D), or FSH plus DETA/NO plus ODQ (FSH+D+O). Chen. Effects of NO and cGMP on inhibin A. Fertil Steril 2003. Fertility and Sterility 2003 79, 687-693DOI: (10.1016/S0015-0282(02)04823-9)

FIGURE 6 Northern blot analysis of inhibin α and βA subunit mRNA levels in granulosa cells treated with media alone (C), FSH (F), F plus DETA/NO (F+D), or F plus dibutyryl cGMP (F+cG). The migrating positions of the 28S and 18S ribosomal RNAs are shown. Chen. Effects of NO and cGMP on inhibin A. Fertil Steril 2003. Fertility and Sterility 2003 79, 687-693DOI: (10.1016/S0015-0282(02)04823-9)

FIGURE 7 Slot blot analysis of inhibin α (top) and βA (bottom) subunit mRNA levels. Immature granulosa cells were treated as in Figure 5. Total RNA was analyzed using 32P-labeled inhibin subunit riboprobes, and results were normalized for 28S rRNA levels. Each value represents the mean of six separate determinations. Chen. Effects of NO and cGMP on inhibin A. Fertil Steril 2003. Fertility and Sterility 2003 79, 687-693DOI: (10.1016/S0015-0282(02)04823-9)