Neurosteroids: steroidogenesis in primary cultures of rat glial cells after release of aminoglutethimide blockade.
Newborn rat glial cells were established in primary culture for greater than or equal to 3 weeks under conditions previously reported to permit differentiation of cholesterol side-chain cleavage activity. The cells were incubated for 48h with 3-H-Mevalonolactone in presence of Mevinolin (avoiding isotopic dilution of endogenous origin) and aminoglutethimide, a blocker of cholesterol side-chain cleavage. Radioactive steroid synthesis was then allowed to proceed in absence of aminoglutethimide, for 16h in presence of Trilostane, an inhibitor of 3 beta-hydroxysteroid dehydrogenase precluding formation of delta 4-3-ketosteroids. 3H-cholesterol accounted for approximately 10 percent of the radioactivity in cell extracts. 3H-delta 5-pregnene-3 beta, 20 alpha-diol (20-OH P) was the major steroid produced and was released in the culture medium. Dexamethasone (10 nM) increased 20-OH P formation by 30 percent, whereas cellular 3H-cholesterol decreased more than expected from the augmented formation of 20-OH P.
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Lab. Hormones, INSERM U33, Bicêtre, France.
Jung-Testas
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Robel
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Baulieu
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D000616:Aminoglutethimide / Q000494:pharmacology; D000818:Animals; D002478:Cells, Cultured; D002784:Cholesterol / Q000096:biosynthesis; D003907:Dexamethasone / Q000494:pharmacology; D066298:In Vitro Techniques; D008798:Mevalonic Acid / Q000378:metabolism; D009457:Neuroglia / Q000378:metabolism*; D011284:Pregnenolone / Q000031:analogs & derivatives / Q000096:biosynthesis; D051381:Rats; D013256:Steroids / Q000096:biosynthesis*
D013256:Steroids; D000616:Aminoglutethimide; D011284:Pregnenolone; D003907:Dexamethasone; C009033:5-pregnene-3,20-diol; D002784:Cholesterol; D008798:Mevalonic Acid