Biol Reprod
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Biology of Reproduction, Vol 51, 465-471, Copyright © 1994 by Society for the Study of Reproduction

ARTICLES

Steroidogenic cytochrome P450 enzyme messenger ribonucleic acids and follicular fluid steroids in individual follicles during preovulatory maturation in the pig

HD Guthrie, JA Barber, JK Leighton and JM Hammond
U.S. Department of Agriculture, Livestock and Poultry Science Institute, Beltsville, Maryland 20705.

Changes in follicular concentrations of steroidogenic cytochrome P450 enzyme mRNAs were determined during preovulatory maturation. RNA was isolated from 59 individual follicles dissected from 18 pigs during altrenogest-synchronized preovulatory follicular maturation: at Day 1 (pre-follicular phase), Day 3 (early follicular phase), Day 5 (mid- follicular phase), and Day 7 (late follicular phase, 24-36 h after the onset of the LH surge). Follicular fluid was aspirated for steroid RIA. RNA was also isolated from pooled granulosa cells, theca tissue, and luteal tissue. RNA was analyzed by Northern and slot-blot procedures using cDNA probes to human aromatase cytochrome P450 (P450arom), porcine 17 alpha-hydroxylase cytochrome P450 (P450(17) alpha), and porcine cholesterol side-chain cleavage cytochrome P450 (P450scc). P450arom mRNA was expressed in both granulosa and theca interna cells but was not detectable in luteal tissue from Day 13 of the estrous cycle. Follicular P450arom mRNA concentration tended to increase between Days 1 and 5 and decreased (p < or = 0.05) by 92% between Days 5 and 7. Follicular fluid estradiol-17 beta concentration increased 17- fold between Days 1 and 5 and then decreased (p < or = 0.05) by 96% between Days 5 and 7. P450(17) alpha mRNA was present in theca interna but was not detected in granulosa cells or luteal tissue. Follicular P450(17) alpha mRNA concentrations did not differ significantly among days, but the content per follicle increased (p < 0.05) between Days 1 and 5 and decreased (p < 0.05) between Days 5 and 7.(ABSTRACT TRUNCATED AT 250 WORDS)




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Copyright © 1994 by the Society for the Study of Reproduction.