Comparison of subpopulations of luteal cells obtained from cyclic and superovulated ewes

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Abstract
Peripheral blood samples were collected daily (Days 1-10 after ovulation) and analysed for progesterone content. Luteal tissue was collected on Day 10 after the LH surge, or Day 10 after hCG injection from cyclic and superovulated ewes, respectively. The tissue was enzymically dispersed and an aliquant was utilized for measurement of cell diameters, and staining for 3.beta.-hydroxy-.delta.5-steroid dehydrogenase-.delta.5,.delta.4-isomerase activity (3.beta.-HSD). The remaining cell preparation was separated into small (10-22 .mu.m) and large (> 22 .mu.m) cell fractions by elutriation. Small and large cell suspensions were incubated (37.degree.C, 2 h) in the presence or absence or ovine LH (100 ng/ml) or dbcAMP (2 mM) and progesterone content of the medium was measured. Superovulation did not effect circulating progesterone concentrations, when expressed per mg luteal tissue recorded; basal progesterone production by small or large luteal cells; the unresponsiveness of large luteal cells to ovine LH or dbcAMP; the ratio of small:large cells recovered by dissociation the mean diameter of total cells; or the mean diameter of large cells. However, the mean cell diameter and LH stimulation of progesterone production by small cells were greater (P < 0.05) in luteal tissue collected from superovulated than that from cyclic ewes. These differences appear to be an amplification of basic function. Therefore, we conclude that corpora lutea obtained from superovulated ewes can be used to study functional aspects of small and large cells.