Receptor mediated gonadotropin action in gonadal tissues: relationship between blood cholesterol levels and gonadotropin stimulated steroidogenesis in isolated rat Leydig and luteal cells.

The present studies were performed to evaluate the role of steroid precursors and plasma lipoproteins in gonadal tissue steroidogenesis. Leydig cell suspension isolated from rat testes responded to hCG. Bt2cAMP, 8 Br-cAMP and cholera toxin with an increase in testosterone response. Administration of 4-aminopyrazolo[3,4-d]pyrimidine (4-APP) reduced the plasma cholesterol and testosterone levels in a time and dose dependent manner. This treatment also reduced the steroidogenic capacity of isolated Leydig cells both under basal conditions and in response to trophic hormone. Different doses of 4-APP up to 25 mg/kg BW and up to 4 days of treatment, however, did not modulate cholesterol and cholesterol ester contents of isolated Leydig cells. 4-APP treatment also had no effect on testis weight, phospholipid content, protein synthesis and energy metabolism in isolated Leydig cells. Similarly, administration of 4-APP (12.5 mg/kg) to PMSG-hCG primed rats beginning on day 3, post hCG, drastically reduced the circulating cholesterol and progesterone levels. Injection of the drug also produced an inhibition in vitro luteal cell steroidogenesis and a reduction in cellular cholesterol esters and free cholesterol contents. Addition of LDL or HDL to incubation medium reversed the inhibitory effect of 4-APP on luteal cell steroidogenesis while this inhibition persisted in Leydig cells. Injection of rats with Triton-WR-1339 (mg/kg BW) resulted in a 10-fold increase in plasma cholesterol and a contrasting decrease in testosterone levels. This treatment, however, produced no effect on in vitro Leydig cell steroidogenesis or cellular content of cholesterol esters and free cholesterol. It appears that the Leydig and luteal cells process and utilize lipoprotein-delivered cholesterol for steroidogenesis through different mechanism(s). These studies thus demonstrate differential actions and an acute regulatory role of lipoproteins in gonadotropin modulated steroidogenesis in two different gonadal tissue.