Calcium-, phospholipid-dependent protein kinase activity of cultured rat Sertoli cells and its modifications by vitamin A

The activity of the calcium-, phospholipid-dependent protein kinase (PKc) was partially characterized in Sertoli cell cultures prepared from 20-day-old rats. The calcium dependency, the requirements for phosphatidylserine and diolein, as well as the Km for ATP and for the tumor promoter TPA, were determined in total cell extracts. The specific activity of PKc was almost 3-fold higher in the soluble than in the particulate fraction of Sertoli cells. Treatment of cultured Sertoli cells with retinol inhibited, within 1 h of treatment, both the soluble and the particulate fraction-associated PKc activity, with an IC50 of 0.1 microM. Partial inhibition of PKc activity was obtained treating Sertoli cell cultures with FSH, while testosterone was ineffective. However, both FSH and testosterone potentiated the inhibitory effect of retinol. Less differentiated Sertoli cells, obtained from 8-day-old rats, displayed higher PKc activity and a pattern of subcellular distribution of the enzyme opposite to that of Sertoli cells obtained from 20-day-old rats. These data suggest that the actual PKc activity of rat Sertoli cells be negatively regulated by retinol and, spontaneously, during the progression of Sertoli cell differentiation.