Modulatory action of epidermal growth factor on differentiated human granulosa lutein cells: cross-talk between ligand activated receptors for EGF and gonadotropin

Abstract

A number of local regulatory factors including polypeptide growth factors like epidermal growth factor (EGF) have been suggested to play an active role within the human ovary. In order to understand the physiology of EGFs action, it is essential to demonstrate and characterize the receptors for this growth factor on ovarian cells which was the aim of this study. We demonstrate using [ 125I]EGF that specific high affinity binding sites with K a for this ligand reaching 2.2 × 10 −9 M for growing cultures of human granulosa-lutein cells and 0.13 × 10 −9 M for the membrane fraction prepared from these cells. Additionally we have identified a 170 kD protein as the EGF receptor with the help of affinity cross linking and immunoblotting procedures. Furthermore, we observed that a pretreatment of granulosa lutein cells with EGF for a short duration (0–30 min) leads to a dose-and time dependent upregulation of the LH-receptor-coupled adenylate cyclase activity. A maximal effect (159 ± 12% increase compared with untreated cells, P < 0.001, n = 4) was reached at 10–15 min with 10–20 ng/ml EGF. Specific inhibition of the receptor tyrosine kinase activity abolished the observed EGF-induced sensitization of the cyclase activity. Differentiation of granulosa cells in vivo is a prerequisite for ovulation and later transformation into highly differentiated lutein cells, a process depending on the presence of ligands that elevate cAMP production. The observed modulation of the adenylate cyclase by EGF could be a regulatory component for the differentiated status of the granulosa cells during different phases of the cycle.