The Role of Luteinizing Hormone and Prolactin in the Regulation of Insulin Receptors in Leydig Cells of the Adult Rat

Abstract

The role of luteinizing hormone (LH), prolactin and their combination in the regulation of insulin receptors in Leydig cells was studied. Leydig cells were isolated from adult male Wistar rats and measurement of insulin binding and internalization was done by incubating the cells with a saturating concentration of 125I‐insulin in the presence or absence of different doses of unlabeled LH/insulin. LH exposure (100 and 200 ng dose) caused a significant increase in Leydig cell surface and internalized insulin receptor concentrations. Prolactin at all doses was ineffective in inducing a significant change in insulin receptor concentration. Under basal condition, Leydig cell surface binding of 125I‐insulin was greater than the internalization at 34°C but at 4°C, surface binding remained lower than that at 34°C with negligible internalization. Internalization of insulin receptors was measured by incubating the cells at 4°C for 16 h and then rapidly incubating at 34°C for various time intervals (60, 120, and 180 min). LH/PRL or LH + PRL did not induce any significant change in the internalization of 125I‐insulin at 60 and 120 min. The rate of internalization was greater at 120 min in basal as well as LH/PRL exposed Leydig cells, compared to 60 min of incubation. Prolactin alone did not evoke any appreciable change in internalization of 125I‐insulin compared to basal at all three time points tested. Total and acid soluble release of 125I‐insulin recorded a significant increase in Leydig cells exposed to LH, which was marginally potentiated when prolactin was added along with LH. Monensin treatment of Leydig cells prevented the recycling of insulin receptors to the cell surface and thereby suppressed the surface binding and enhanced the internalized 125I‐insulin. Under cycloheximide treatment, neither surface bound nor internalized 125I‐insulin recorded a significant change compared to their respective basal values. It is concluded from the present study that LH has dose‐dependent biphasic effects on insulin receptors in Leydig cells by modulating the internalization and intracellular processing of hormone–receptor complexes but prolactin has no such effects.