The cytosolic Ca2+ concentration in acutely dissociated subfornical organ (SFO) neurons of rats: Spontaneous Ca2+ oscillations and Ca2+ oscillations induced by picomolar concentrations of angiotensin II

Abstract

Characteristics of subfornical organ (SFO) neurons were examined by measuring the cytosolic Ca2+ concentration ([Ca2+]i) in acutely dissociated neurons of the rat. SFO neurons, defined by the responsiveness to 50 mM K+ (n = 67) responded to glutamate (86%), angiotensin II (AII) (50%), arginine vasopressin (AVP) (66%) and/or carbachol (CCh) (61%), at their maximal concentrations, with marked increases in [Ca2+]i. More than a half (174/307) of SFO neurons examined exhibited spontaneous Ca2+ oscillations, while the remainder showed a relatively stable baseline under unstimulated conditions. Spontaneous Ca2+ oscillations were suppressed when extracellular Ca2+ was removed and were inhibited when extracellular Na+ was replaced with equimolar N-methyl-D-glucamine. Ca2+ oscillations were unaffected by the inhibitor of Ca2+-dependent ATPases cyclopiazonic acid, the N-type Ca2+ channel blocker ω-conotoxin GVIA and the P/Q-type Ca2+ channel blocker ω-agatoxin IVA, but significantly inhibited by the high-voltage-activated Ca2+ channel blocker Cd2+ and the L-type Ca2+ channel blocker nicardipine. Ca2+ oscillations were also completely arrested by the voltage-gated Na+ channel blocker tetrodotoxin in 50% of SFO neurons but only partially in the remaining neurons. These results suggest that SFO neurons exhibit spontaneous membrane Ca2+ oscillations that are dependent in part on Ca2+ entry through L-type Ca2+ channels, whose activation may result from burst firing. Moreover, AII at picomolar concentrations induced Ca2+ oscillations in neurons showing no spontaneous Ca2+ oscillations, while spontaneous Ca2+ oscillations were arrested by gamma-aminobutyric acid (10 μM), suggesting that rises in [Ca2+]i during Ca2+ oscillations may play an important role in the modulation of SFO neuron function.