[3-Me-His(2)]-TRH combined with dopamine withdrawal rapidly and transiently increases pyroglutamyl aminopeptidase II activity in primary cultures of adenohypophyseal cells

Abstract

TRH is hydrolyzed by pyroglutamyl aminopeptidase II (PP II), a highly specific ecto-enzyme which is localized on the surface of lactotrophs. To study whether PP II activity may be rapidly regulated during a burst of prolactin secretion, we used an in vitro model in which primary cultures of adenohypophyseal cells were incubated with 500 nM dopamine (DA) for 24 h prior to treatments. We observed a rapid increase of PP II activity when 100 nM [3-Me-His(2)]-TRH, a TRH agonist, was added at removal of DA. PPII activity was maximal after 20 min of treatment and reduced to time 0 activity at 30 min. Dopamine withdrawal alone, slightly and transiently, modified the enzyme activity: an initial activation at 15 min was followed by a transient inhibition at 20 min. The specific contribution of [3-Me-His(2)]-TRH in this paradigm was a transient enhancement of PP II activity. If DA was not removed, [3-Me-His(2)]-TRH was ineffective. These data demonstrate that during in vitro conditions that mimic a suckling episode, adenohypophyseal PP II activity is rapidly and reversibly adjusted.