Control of progesterone production in small and large bovine luteal cells separated by flow cytometry.

摘要: Corpora lutea were collected from Holstein heifers on Days 10 and 12 of the oestrous cycle cells dispersed with collagenase. The separated into preparations highly purified (90-99%) small (less than 20 microns) large (greater 25 luteal by unit gravity sedimentation fluorescence-activated cell sorting. Net progesterone accumulation 1 x 10(5) 10(3) during 2 4 h incubations, respectively, measured after additions LH, PGF-2 alpha, phorbol esters, alone in combination. Progesterone synthesis was increased (P less 0.05) dibutyrate (PBt2) or alpha small, but not large, (10.1 +/- 3.0 18.1 5.0 ng/10(5) for 0 50 nM-PBt2, 19.9 3.2 44.2 9.3 microgram alpha/ml). previously reported stimulatory effects PKC activation addition to total are therefore entirely attributable theca-derived cells. Small responded low levels LH (9.1 1.1, 69.0 5.4 154.7 41.4 0, 5 ng LH/ml, P 0.05), while only high (1635 318, 2662 459 3386 335 pg/10(3) 100 1000 0.05). inhibited LH-, 8-Br-cAMP- forskolin-stimulated (3052 380, 3498 418, 3202 391 0.5 mM-8-Br-cAMP, microM-forskolin respectively 1750 487, 2255 468, 2165 442 + 8-Br-cAMP forskolin, respectively), indicating that inhibitory effect occurs at a site distal cAMP generation. These results suggest targets luteolytic responsible luteotrophic vitro.