Vasoactive intestinal peptide regulates cholesterol side-chain cleavage cytochrome P-450 (P-450scc) gene expression in granulosa cells from immature rat ovaries.
Vasoactive intestinal peptide (VIP), a neuropeptide present in ovarian nerves, has been previously shown to induce synthesis of the side-chain cleavage cytochrome P-450 enzyme which catalyzes the conversion of cholesterol to pregnenolone (the rate-limiting step in progesterone synthesis). In the present study we demonstrate, by means of a bovine 3'-specific P-450scc cDNA probe, that this VIP effect is exerted at least partially at the level of gene expression in cultured granulosa cells that were isolated from estrogen-primed, immature rats. The size and level of the 2.0 kilobase P-450scc mRNA species was assessed by Northern blot analysis, while the translatability of this mRNA was assayed by immunoisolation of the 35S-labeled P-450scc precursor protein translated from total RNA of control and stimulated granulosa cells. FSH was much more effective than VIP at increasing P-450scc mRNA concentrations in cultured granulosa cells, whereas secretin treatment was ineffective. The results suggest that, like FSH, the stimulatory effect of the neuropeptide VIP on ovarian progesterone secretion involves regulation of P-450scc gene expression during functional maturation of the prepubertal ovary.
Trzeciak
W H
WH
Department of Biochemistry, Cecil H and Ida Green Center for Reproductive Biology Sciences, University of Texas Health Science Center Dallas 75235.
Waterman
M R
MR
Simpson
E R
ER
Ojeda
S R
SR
D000818:Animals; D015152:Blotting, Northern; D002474:Cell-Free System; D002478:Cells, Cultured; D002786:Cholesterol Side-Chain Cleavage Enzyme / Q000096:biosynthesis*; D005260:Female; D015971:Gene Expression Regulation, Enzymologic / Q000502:physiology*; D006107:Granulosa Cells / Q000201:enzymology*; D014176:Protein Biosynthesis / Q000502:physiology; D012313:RNA / Q000302:isolation & purification; D051381:Rats; D012741:Sexual Maturation / Q000502:physiology*; D014660:Vasoactive Intestinal Peptide / Q000502:physiology*
D014660:Vasoactive Intestinal Peptide; D012313:RNA; D002786:Cholesterol Side-Chain Cleavage Enzyme