FSH STIMULATION OF3H-GLUCOSAMINE-INCORPORATION INTO PROTEOGLYCANS BY PORCINE GRANULOSA CELLS IN VITRO*

Abstract

Porcine granulosa cells from small follicles were incubated in the presence of 1 μCi/ml ³H-glucosamine and various hormones. Proteoglycans were digested with protease, and the mucopolysaccharides (MPS) were recovered. Most of the labeled MPS was found in secretory products. Maximum incorporation of ³H-glucosamine into MPS occurred after incubation with FSH for 8–16 h. Cell concentrations of 0.5–4.0 x 10⁶/ml proved to be ample. Highly purified porcine or human FSH gave consistent, significant stimulation by doses in the order of 100 pg/ml. Linear log-dose response curves occurred from 0.1 to 10.0 ng/ml, and maximal stimulation at 10 ng/ml was 22-fold above control. Preparations of hCG, hTSH and NIH-LH-S18 were not effective. Incubations with 1–10 mM dibutyryl cyclic AMP significantly increased incorporation into MPS, suggesting that effects of FSH may be mediated by cyclic AMP.