Activation of extracellular signal-regulated kinase (ERK) and induction of mitogen-activated protein kinase phosphatase 1 (MKP-1) by perifused thyrotropin-releasing hormone (TRH) stimulation in rat pituitary GH3 cells.

We investigated the pattern of extracellular signal-regulated kinase (ERK) phosphorylation and the induction of mitogen-activated protein kinase phosphatase 1 (MKP-1) by thyrotropin-releasing hormone (TRH) under various stimulation conditions in pituitary GH3 cells. In static culture, ERK activation by continuous TRH was maximal at 10 min and persisted for up to 60 min, with a return to the basal level by 2h. Stimulation with continuous TRH in perifused cells resulted in a similar level of ERK phosphorylation. MKP-1 was expressed 60 min following either static or perifused, continuous TRH stimulation. When cells were stimulated with pulsatile TRH every 30 min, ERK activation was maximal at 10 min and returned to its baseline level by 30 min. ERK was phosphorylated again with each subsequent pulse. Pulsatile TRH did not induce MKP-1. Prolactin promoter activity following continuous, static TRH stimulation was higher than that following perifused TRH stimulation. TRH at a frequency of one pulse every 30 min increased prolactin promoter activity similar to that of perifused, continuous TRH stimulation. Additionally, changes in pulse frequency resulted in alterations in the level of prolactin promoter. Following static stimulation, a 10 min exposure to TRH was sufficient to obtain full activation of the prolactin promoter. Additionally, a 5-10 min exposure of TRH was sufficient to maintain ERK activation. A single 5-min pulse of TRH stimulation resulted in low activation of the prolactin promoter. ERK activation was necessary for prolactin gene transcription; however, prolactin gene transcription is not entirely determined by the strength or duration of TRH-induced ERK activation.