Follicle-stimulating hormone regulation of cytochrome P-450 side-chain cleavage messenger ribonucleic acid accumulation by porcine granulosa cells isolated from small and medium follicles.
The experiments described here were conducted to examine regulation of cytochrome P-450 side-chain cleavage (SCC) mRNA accumulation in porcine granulosa cells isolated from small (1-4-mm) and medium (5-6-mm) follicles. Granulosa cells were cultured under the following conditions: 1) for 48 h or 96 h with 0, 50, or 200 ng/ml porcine FSH; 2) for 96 h with 200 ng/ml FSH and aminoglutethimide (100 microM); and 3) for 96 h with forskolin (100 microM). Total RNA was extracted and examined by Northern and dot-blot hybridization analysis, and culture media were assayed for progesterone concentration. Northern blot analysis revealed a single band approximately 2.1 kb in size. Accumulation of SCC mRNA by granulosa cells was both FSH dose- and culture time-dependent (p less than 0.05) with maximal increases approximately 4.5 times control levels. Aminoglutethimide reduced progesterone production by about 80% while having no effect on granulosa cell accumulation of SCC mRNA compared to cells stimulated with 200 ng/ml of FSH. Forskolin-treated cells produced significantly more progesterone than did cells treated with FSH, but accumulation of SCC mRNA was similar. In response to FSH, concentration of SCC mRNA did not vary with follicle size, but granulosa cells from small follicles produced significantly more progesterone than did those from medium follicles. These results demonstrate that concentration of SCC mRNA in cultured porcine granulosa cells is FSH dose-dependent, does not vary significantly in cells from small- and medium-sized follicles, and is correlated with progesterone production, but may not parallel progesterone secretion. This last observation indicates that control at sites other than SCC mRNA can affect progesterone production.
Mulheron
G W
GW
U.S. Department of Agriculture, Roman L. Hruska U.S. Meat Animal Research Center, Clay Center, Nebraska 68933-0166.
Wise
T H
TH
Stone
R T
RT
Ford
J J
JJ
D000616:Aminoglutethimide / Q000494:pharmacology; D000818:Animals; D002469:Cell Separation; D002786:Cholesterol Side-Chain Cleavage Enzyme / Q000235:genetics* / Q000378:metabolism; D005576:Colforsin / Q000494:pharmacology; D015342:DNA Probes; D004305:Dose-Response Relationship, Drug; D005260:Female; D005640:Follicle Stimulating Hormone / Q000502:physiology*; D006107:Granulosa Cells / Q000187:drug effects / Q000378:metabolism*; D015151:Immunoblotting; D006080:Ovarian Follicle / Q000166:cytology*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism*; D013552:Swine / Q000378:metabolism*; D013997:Time Factors
D015342:DNA Probes; D012333:RNA, Messenger; D000616:Aminoglutethimide; D005576:Colforsin; D005640:Follicle Stimulating Hormone; D002786:Cholesterol Side-Chain Cleavage Enzyme