Arachidonic acid elevates cytosolic free calcium concentration in rat anterior pituitary cells

Arachidonic acid is liberated from phospholipids by various hypothalamic releasing hormones and may be involved in stimulus-secretion coupling in rat adenohypophysis. In the present study, the effect of exogenous arachidonic acid on calcium homeostasis in rat anterior pituitary cells was investigated in vitro. Arachidonic acid markedly stimulated the release of various anterior pituitary hormones (beta-endorphin, luteinizing hormone, growth hormone). Arachidonic acid (10 mumol/l) decreased the initial rate of 45Ca2+ uptake. In cells prelabelled with 45Ca2+, arachidonic acid (10 mumol/l) decreased the exchangeable cell calcium content and increased the rate of 45Ca2+ extrusion. Cytosolic free calcium concentration [( Ca2+]i) was measured with the fluorescent indicator fura-2. Arachidonic acid markedly elevated [Ca2+]i. The concentration dependency of this effect (1 mumol/l and above) was similar to that on hormone secretion. Arachidonic acid (6 mumol/l) elevated [Ca2+]i by about 300 nmol/l, and arachidonic acid (10 mumol/l) raised [Ca2+]i into the micromolar range. The effect of arachidonic acid (3 mumol/l) on [Ca2+]i was not influenced by inhibitors of arachidonic acid metabolism (nordihydroguaiaretic acid, BW755C). In Ca2+-free media (Ca2+ omitted, EGTA 2 mmol/l), the effect of arachidonic acid (3 mumol/l) on [Ca2+]i was almost unimpaired, whereas the effect of arachidonic acid (10 mumol/l) was reduced. Thus, the secretagogue arachidonic acid induces calcium mobilization and an increase in cytosolic free calcium concentration. These actions further qualify arachidonic acid as a potential intracellular mediator of stimulus-induced hormone secretion from rat adenohypophysis.