Effects of vasopressin on isolated rat adrenal chromaffin cells

It has been demonstrated that arginine vasopressin (AVP) is synthesized not only in specific hypothalamic nuclei, but also in the adrenal medulla where it is thought to regulate adrenal functions by autocrine and paracrine mechanisms. In order to further characterise the effects of AVP on rat adrenal chromaffin cells, we examined: (a) the mRNA expression for V_1a and V_1b AVP receptors in these cells; (b) the effects of AVP on the membrane potential and membrane currents measured with the whole-cell patch-clamp technique; and (c) effect of AVP on catecholamine release from single adrenal chromaffin cells measured with carbon fibre microelectrodes. Reverse transcription-polymerase chain reaction (RT-PCR) on tissue punch samples obtained from the adrenal medulla demonstrated message for both the V_1a and V_1b receptors, while material obtained from the adrenal cortex showed expression of the V_1a receptor only. Single-cell RT-PCR conducted on acutely isolated chromaffin cells showed message for the V_1a receptor in 84% of cells, while 38% of cells also contained message for the V_1b receptor (n=45). Under current-clamp recording, responses to AVP application (4-40 μM) were variable; 22/34 (65%) tested cells were depolarised, 29% hyperpolarised, and the remaining cells showed a biphasic response. Changes in membrane potential of either direction were dose-dependent and accompanied by a decrease in cell membrane resistance. Under voltage-clamp (V_hold=-60 mV), AVP evoked inward current in 27/52 (52%) and outward current in 16/52 (31%) chromaffin cells. Both types of AVP-evoked responses were blocked by co-application of a nonselective V_1a/V_1b antagonist. Application of AVP evoked prolonged bursts of amperometric currents (indicative of catecholamine release) in 4/9 tested cells, but reduced the currents evoked by ACh application in all tested cells (n=7). These findings demonstrate a complex action of AVP on adrenal chromaffin cells, with individual adrenal chromaffin cells responding with either excitation or inhibition. This response pattern may be related to the expression of V₁ receptor subtypes.