Arachidonic acid affects membrane ionic conductances of GH3 pituitary cells

Arachidonic acid (AA) stimulates prolactin release from pituitary cells, by mechanisms not yet understood. In this work, we analyzed the effects of AA on membrane ionic conductances in a clonal line of anterior pituitary cells (GH3/B6), finding time- and dose-dependent effects of AA on their membrane ionic conductances. The predominant response at concentrations between 100 nM and 10 microM was a prolongation of the action potential (AP) and an increase in the transient after-hyperpolarization potential. Voltage clamp studies showed that this was associated with a decrease in a voltage-dependent potassium current and an increase in a voltage-dependent calcium current. In some cells (30%) the effect of AP duration was less important, but spike firing was enhanced. For the highest concentrations used (1 and 10 microM) the effects described above were preceded by hyperpolarization of the cell membrane; in voltage clamp it was shown that this hyperpolarization resulted from the activation of a calcium-dependent potassium conductance suspected to be due to the release of intracellular calcium. The calcium store affected by AA was, at least in part, insensitive to vanadate and heparin. These data suggest that AA may enhance intracellular calcium concentration by increasing calcium entry during each voltage-dependent calcium AP, by increasing the spike frequency, or by releasing calcium from an intracellular compartment. The resulting rise in cytosolic free calcium concentration may be a key link in the process by which AA stimulates prolactin release in GH3/B6 pituitary cells.