Protein kinase C inhibits TRH-stimulated phosphoinositide hydrolysis in GH3 cells

The GH3 pituitary tumor line expresses TRH receptors that stimulate phosphoinositide hydrolysis and hormone secretion. After protein kinase C was identified in GH3 cells by direct labeling with [3H]phorbol dibutyrate (PDB), the response to phorbol ester and TRH pretreamtent on subsequent TRH-stimulated inositol phosphate (IP) accumulation was found to be inhibitory. Both phorbol myristate acetate (PMA) and PDB were effective in the regard at low nM concentrations within a few minutes, whereas phorbols that do not stimulate protein kinase C were without effect. Furthermore, the mono-, bis- and tris-phosphate forms of IP were all reduced by an average of 30-40% after 5 min of PMA. TRH concentration-response studies indicated a clear change in TRH efficacy induced by PMA. Finally, preincubation with TRH itself was also capable of reducing the subsequent response to TRH. Because TRH receptor action is thought to the activate protein kinase C by producing diacylglycerol, these data indicate a negative feedback system via protein kinase C operative during continuous exposure to TRH in GH3 cells.