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University of Washington, Department of Physiology and Biophysics, Seattle 98195, USA.

1. Indo-1 microfluorimetry and patch clamp techniques were used to study the decrease in cytosolic [Ca2+] ([Ca2+]i) caused by dopamine (D2) receptor activation and the calcium dependence of membrane capacitance changes in single rat melanotrophs. 2. [Ca2+]i decreased when extracellular calcium was removed or when the calcium channel blockers nickel (2 mM) or cadmium (100 microM) were applied by bath perfusion. 3. Quinpirole, a dopamine (D2) receptor agonist, reduced [Ca2+]i by 55 +/- 9 nM and hyperpolarized membrane potential by 29 +/- 9 mV simultaneously. 4. Quinpirole-induced [Ca2+]i decrease required deactivation of voltage-dependent calcium channels. Voltage clamping the membrane potential at -25 mV prevented the quinpirole-induced [Ca2+]i decrease. Nickel (2 mM) reduced [Ca2+]i without hyperpolarization and precluded additional [Ca2+]i decrease by quinpirole. 5. Membrane capacitance measurement of secretion rates in cells dialysed with buffered calcium solutions showed that secretion began at approximately 400 nM Cai2+. 6. Melanotrophs have IP3-sensitive calcium stores, but no caffeine-sensitive calcium stores. Calcium released from IP3-sensitive calcium stores also stimulated secretion. 7. Secretion in melanotrophs is modulated by protein kinase activators. cAMP (200 microM) enhanced secretion at [Ca2+]i > 1000 nM. Phorbol myristate acetate (PMA; 200 nM) enhanced secretion at [Ca2+]i < 400 nM, but not in the absence of calcium. 8. Dopamine receptor activation can reduce secretion by reducing the calcium influx through calcium channels with hyperpolarization of the membrane potential. However downregulation of either cAMP or protein kinase C activity may also contribute to the decrease in secretion.