Androstenedione and progesterone production in vitro by the inner or the outer theca cells in preovulatory follicles of gonadotropin stimulated calves

This study reports some of the steroidogenic characteristics of the interna and externa theca cells taken from young and eCG primed calves. These cells were isolated from large healthy follicles. They were separately cultured for 3 days in absence or in presence of steroid substrates. Androstenedione (A4) and progesterone (P4) were measured by radioimmunoassay (RIA). In control conditions, A4 levels, higher in interna than in externa cells (**P<0.01), decreased during cultures (**P<0.01). In both cell types, A4 increased in presence of 17α-hydroxypregnenolone (17OHP5), pregnenolone (P5) and 22R-hydroxycholesterol (22R-chol) (*P<0.05) but not with P4 or 17α-hydroxyprogesterone (17OHP4) (P>0.05). The most efficient substrate was dehydroepiandrosterone (DHEA) (*P<0.05). In control conditions, P4 levels increased in both cell types. They were higher in externa than in interna cells on day 1, the reverse was observed on day 3. P4 levels increased after addition of 22R-chol and P5 (*P<0.05) but not with 17OHP5, 17OHP4 and DHEA (P>0.05) from day 1 in externa cells and only on day 3 in interna cells. P4 levels measured on day 1 were lower than the quantity of P4 added as a substrate. These results, obtained with theca cells from young calf follicles, indicate: 1/A4 is synthesized by the Δ5 pathway and 17α-hydroxylase activity decreases in vitro, 2/externa and interna cells differ by the quantities of A4 and P4 produced, 3/both lack precursors to produce A4 and P4 but their 3β-hydroxysteroid dehydrogenase activity subsists, 4/P4 could be metabolized during the first 2 days in both cell cultures.