Delayed activation of phospholipase D by gonadotropin-releasing hormone in a clonal pituitary gonadotrope cell line (αT3-1)

Stimulation of cultured pituitary cells from a gonadotrope lineage (αT3-1) by the gonadotropin-releasing hormone agonist analog [D-Trp 6]GnRH (GnRH-A) resulted in a manifold increase in accumulation of phosphatidylethanol, a specific product of phospholipase D phosphatidyl transferase activity when ethanol is the phosphatidyl group acceptor. Levels of the natural lipid product of phospholipase D, phosphatidic acid, were increased 2–3-fold. Activation of phospholipase D by GnRH-A was dose- and time-dependent and was blocked by a GnRH receptor antagonist [D-pClPhe 2,D-Trp 3.6]GnRH. GnRH-A stimulated phospholipase D activity after a lag of 1–2 min. We conclude that in αT3-1 gonadotropes GnRH receptor occupancy results in delayed activation of phospholipase D which could participate in late phases of gonadotrope regulation by the neurohormone.