Steroidogenic activities in MA-10 Leydig cells are differentially altered by cAMP and Müllerian inhibiting substance.
In addition to causing Müllerian duct regression in fetal males, Müllerian inhibiting substance (MIS) inhibits the expression of the bifunctional cytochrome P450, C17 hydroxylase/C(17-20) lyase (Cyp17), the enzyme that catalyzes the committed step in sex steroid synthesis. To investigate the paracrine effects of MIS on steroidogenic activity, we have performed assays with microsomes from mouse MA-10 Leydig cells. With microsomes from untreated MA-10 cells, progesterone was largely metabolized by 5alpha-reductase and subsequently converted by 3-keto steroid reductases to allopregnanolone and epiallopregnanolone. Addition of cAMP to the cells shifted microsomal steroid production to the Cyp17 product androstenedione and its 5alpha,3beta-reduced form, epiandrosterone. Microsomes from MIS-treated cells were less active with the progesterone substrate than those of untreated cells but co-treatment of the cells with both MIS and cAMP mitigated the cAMP-induced shift of the microsomes to androstenedione production. Quantitative analyses of steroid production by Cyp17 showed that cAMP decreased the amount of 17-hydroxyprogesterone produced relative to the androstenedione, suggesting that cAMP signaling lowers the efficiency of the Cyp17 hydroxylase activity or else increases the efficiency of its lyase activity. Addition of MIS to the cAMP-treated cells partially reversed this effect, as well. These results indicate that cAMP induces MA-10 cells to switch from producing 5alpha-reduced progesterone metabolites to producing androstenedione and its metabolites by increasing Cyp17 expression and its relative lyase activity, both of which are inhibited by MIS.
Trbovich
Alexander M
AM
Pediatric Surgical Research Laboratories/WRN 1024, Massachusetts General Hospital and Harvard Medical School, 32 Fruit Street, Boston, MA, USA.
Martinelle
Nina
N
O'Neill
Francis H
FH
Pearson
Elliot J
EJ
Donahoe
Patricia K
PK
Sluss
Patrick M
PM
Teixeira
Jose
J
D019326:17-alpha-Hydroxyprogesterone / Q000378:metabolism; D000735:Androstenedione / Q000096:biosynthesis; D000818:Animals; D054304:Anti-Mullerian Hormone; D000242:Cyclic AMP / Q000494:pharmacology*; D006023:Glycoproteins / Q000494:pharmacology*; D007985:Leydig Cells / Q000187:drug effects* / Q000378:metabolism; D008297:Male; D051379:Mice; D008861:Microsomes / Q000187:drug effects / Q000378:metabolism; D011374:Progesterone / Q000378:metabolism; D051381:Rats; D013254:Steroid 17-alpha-Hydroxylase / Q000187:drug effects; D013256:Steroids / Q000096:biosynthesis*; D013735:Testicular Hormones / Q000494:pharmacology*
D006023:Glycoproteins; D013256:Steroids; D013735:Testicular Hormones; D000735:Androstenedione; D011374:Progesterone; D019326:17-alpha-Hydroxyprogesterone; D054304:Anti-Mullerian Hormone; D000242:Cyclic AMP; D013254:Steroid 17-alpha-Hydroxylase