Effect of high-density lipoprotein on bovine granulosa cells: progesterone production in newly isolated cells and during cell culture

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Abstract
It has been proposed that changes in steroidogenesis which occur during early development of the corpus luteum may be due to increased availability of lipoproteins. Bovine follicular fluid, however, contains significant amounts of high-density lipoprotein (HDL), and granulosa cells are exposed to this lipoprotein before ovulation. To determine whether bovine granulosa cells can utilize HDL the effects of this lipoprotein on freshly isolated, non-luteinized granulosa cells and on granulosa cells undergoing luteinization in serum-free culture were examined. Cells were isolated from non-atretic, antral follicles and cultured for 12 h in 10% (v/v) lipoprotein-deficient serum to allow cell attachment. After this time cells were cultured in serum-free medium. During culture the cells underwent functional luteinization as assessed by an increase in basal progesterone output (9·6-fold in 7 days) which was associated with a marked increase in activity of cholesterol side-chain cleavage and loss of aromatase activity. Dibutyryl cyclic AMP (dbcAMP) increased basal production of progesterone about twofold but HDL alone had no effect. Addition of HDL plus dbcAMP, in contrast, caused a very marked stimulation (up to ten times) of basal steroidogenesis. This trophic effect of HDL and dbcAMP lasted at least 2 weeks. Activity of cholesterol side-chain cleavage was stimulated (threefold over basal) by dbcAMP during culture but HDL was without effect, alone or with dbcAMP. Addition of HDL (in the presence or absence of dbcAMP) to freshly isolated granulosa cells had no significant stimulatory effect on progesterone production over 12 h in six experiments, and in two of these experiments a significant inhibitory effect was seen. Incubation with 22R-hydroxycholesterol, in contrast, caused a marked stimulation of progesterone production, indicating that the steroidogenic capacity of the cells was not already saturated. Results presented here suggest that bovine granulosa cells are able to utilize HDL for steroidogenesis only after luteinization. The massive secretion of progesterone by luteinized granulosa cells which occurs in the presence of HDL suggests that this lipoprotein is very important in the development and maintenance of luteal cell function in cattle. Journal of Endocrinology (1990) 124, 255–260

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