Arachidonic acid (AA) released from membrane phospholipids after activation of surface receptors causes cellular signaling actions in neurons and endocrine cells, including stimulation of prolactin (PRL) release from dissociated rat pituitary cells and clonal cells of the GH3 pituitary tumor line. In the present study, we investigated the effect of exogenous AA on PRL release from dispersed pituitary cells and tried to elucidate the mechanism involved in this process. The effects of AA on cytosolic Ca2+ concentration ([Ca2+]i) were studied using dual-emission microspectrofluorometry in identification lactotrophs and on PRL release in dispersed pituitary cell populations. AA had a dose-dependent effect on [Ca2+]i. At 1 microM, the Ca2+ increase was biphasic: a mobilization of intracellular Ca2+ from intracellular stores was followed by stimulation of Ca2+ influx. For lower concentrations (10 and 100 nM), only the stimulation of Ca2+ influx was observed. AA-induced Ca2+ influx and PRL release were not due to the stimulation of a phorbol 12-myristate 13-acetate-sensitive protein kinase C. In the same way, AA-stimulated PRL release and intracellular Ca2+ increase were independent of intracellular thapsigargin-sensitive Ca2+ pools. Furthermore, blockade of Ca2+ channels suppressed AA-induced PRL release. We hypothesize that Ca2+ influx plays a major role in AA-induced PRL release.

Bradykinin (BK), a nonapeptide, originally discovered in blood, is also present in neurons and fibers of the hypothalamus. We tested the putative releasing factor properties of BK on prolactin (PRL) release from anterior pituitary cells in vitro. BK stimulated the release of PRL in a dose-dependent manner, the threshold concentration being in the range. 0.1–1.0 nM. The release of PRL induced by BKat 1 nM concentration was about 2-fold, delayed and sustained over many minutes. Higher concentrations of BK stimulated PRL release in two phases. The shape of the BK-induced PRL release was superficially similar to that induced by thyrotropin-releasing hormone (TRH). 10 nM BK and 10 nM TRH induced about a 4-fold increase in PRL release within 5 min, followed by a gradual recovery to basal secretion. These results indicate that this peptide can act directly at the anterior pituitary gland to release PRL. Phorbol ester also promoted PRL release over the range of 1–10 nM, but the time course of the release was somewhat different.

Arachidonic acid (AA) has been implicated in signaling actions in several cell types including endocrine cells. In the present study, we investigated the effect of exogenous AA on GH release from dispersed pituitary cells and tried to elucidate the mechanism involved in this process. We show that AA stimulates GH release in a dose- and extracellular calcium-dependent manner. The effects of AA on cytosolic calcium concentration ([Ca²⁺]_i) were studied using dual-emission microspectrofluorimetry in identified somatotropes. AA (1 µM) induced an increase in intracellular calcium concentration ([Ca²⁺]_i) by stimulating Ca²⁺ influx through dihydropyridine-sensitive, voltage-dependent calcium channels. In these cells, the effects of AA were only reduced by the inhibition of protein kinase C (PKC) activity, suggesting that the fatty acid may act by both PKC-dependent and PKC-independent pathways.