Key result
Removal of endogenous nitric oxide by a scavenger reduced c-fos expression, increased c-jun expression, and impaired LH-induced progesterone synthesis in developing porcine granulosa cells.
Transient generation of nitric oxide during maturation of porcine granulosa cells plays a critical role in LH-induced expression of c-fos and c-jun and subsequent progesterone synthesis.
Animal data on nitric oxide in porcine granulosa cells warrant caution; leaves open translational relevance to human ovarian function.
The present study was performed to clarify the synthesis of nitric oxide (NO) and its effect on the expression of transcription factor genes (c-fos, c-jun and ATF-4) during the differentiation of granulosa cells. Granulosa cells prepared from porcine ovarian follicles (1-4 mm diameter) were matured with FSH for 48 h. From 40 h to 48 h of ovine FSH stimulation, nitrite and nitrate increased by 2 folds, and this was accompanied by an increase of cyclic GMP. The cells were exposed to either NO scavenger (carboxy-PTIO: 2-(4-carboxyphenyl)-4, 4, 5, 5-tetramethylimidazoline-1-oxy-3-oxide) or NO donor (NOC18: 2, 2'-(hydroxynitrosohydrazino)bis-ethanamine) before or after NO generation, and further stimulated with ovine LH until 48 h. Removal of endogenous NO (NO scavenger) induced a serious impairment in the LH-induced synthesis of progesterone, whereas NO donor had no significant effect on the synthesis. In the semi-quantitative reverse transcriptase-PCR of the transcription factor mRNAs, removal of endogenous NO resulted in a reduction of c-fos expression, an increase of c-jun and no changes in ATF-4 expression. In contrast, NO donor had no significant effects on the expression of these transcription factor genes. Consequently, the transient generation of NO may have critical roles in the LH-induced expression of transcription factor genes and thereafter transformation of the granulosa cell to the luteal cell.
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Nishida et al. (2000) studied Granulosa cell differentiation. NO scavenger (carboxy-PTIO) or NO donor (NOC18) vs. Control (no treatment) was evaluated on Expression of transcription factor genes (c-fos, c-jun, ATF-4) and progesterone synthesis. Removal of endogenous nitric oxide by a scavenger reduced c-fos expression, increased c-jun expression, and impaired LH-induced progesterone synthesis in developing porcine granulosa cells.
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