Role of Intracellular Na+ in the Regulation of [Ca2+]i in the Rat Suprachiasmatic Nucleus Neurons

Abstract

Transmembrane Ca²⁺ influx is essential to the proper functioning of the central clock in the suprachiasmatic nucleus (SCN). In the rat SCN neurons, the clearance of somatic Ca²⁺ following depolarization-induced Ca²⁺ transients involves Ca²⁺ extrusion via Na⁺/Ca²⁺ exchanger (NCX) and mitochondrial Ca²⁺ buffering. Here we show an important role of intracellular Na⁺ in the regulation of [Ca²⁺]_i in these neurons. The effect of Na⁺ loading on [Ca²⁺]_i was determined with the Na⁺ ionophore monensin and the cardiac glycoside ouabain to block Na⁺/K⁺-ATPase (NKA). Ratiometric Na⁺ and Ca²⁺ imaging was used to measure the change in [Na⁺]_i and [Ca²⁺]_i, and cell-attached recordings to investigate the effects of monensin and ouabain on spontaneous firing. Our results show that in spite of opposite effects on spontaneous firing and basal [Ca²⁺], both monensin and ouabain induced Na⁺ loading, and increased the peak amplitude, slowed the fast decay rate, and enhanced the slow decay phase of 20 mM K⁺-evoked Ca²⁺ transients. Furthermore, both ouabain and monensin preferentially enhanced nimodipine-insensitive Ca²⁺ transients. Together, our results indicate that in the SCN neurons the NKA plays an important role in regulating [Ca²⁺]_i, in particular, associated with nimodipine-insensitive Ca²⁺ channels.

Keywords: Ca2+; Na+; Na+/Ca2+ exchanger; Na+/K+-ATPase; mitochondria; suprachiasmatic nucleus.