PATCH-CLAMP ANALYSIS OF SPONTANEOUS SYNAPTIC CURRENTS IN SUPRAOPTIC NEUROENDOCRINE CELLS OF THE RAT HYPOTHALAMUS

Spontaneous synaptic currents were recorded in supraoptic magnocellular neurosecretory cells using whole-cell patch-clamp techniques in the rat hypothalamic slice preparation. Numerous spontaneous excitatory and inhibitory postsynaptic currents (EPSCs and IPSCs) were observed in the 27 cells recorded. The rate of occurrence of the spontaneous currents and the relative proportion of EPSCs versus IPSCs varied significantly from cell to cell. Miniature EPSCs and IPSCs were clearly distinguished from background noise in TTX (n = 3 cells at 0.5 mug/ml). The frequency of EPSCs and IPSCs decreased by approximately 70% and the largest events were blocked in TTX, but the peaks of the amplitude histograms were shifted by only a few picoamperes. Bicuculline (n = 10 cells at 10 muM and 2 cells at 20 muM) blocked completely all the IPSCs without any detectable effect on the frequency or amplitude of the EPSCs. No slow spontaneous outward currents, indicative of a K+ current from activation of GABA(B) receptors, were observed. The alpha-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid (AMPA)/kainate-type glutamate receptor antagonist 6-cyano-2,3-dihydroxy-7-nitroquinoxaline (CNQX; n = 7 cells at 10 muM) consistently blocked all EPSCs without any apparent effect on the frequency or amplitude of the IPSCs. No synaptic events could be detected when CNQX was applied in combination with bicuculline (n = 4). The decay phase of averaged spontaneous IPSCs and EPSCs recorded at resting membrane potential could be well fitted by single exponential functions in most cells. The time constants ranged from 0.92 to 3.0 msec for EPSCs (five cells) and from 5.3 to 6.6 msec for IPSCs (four cells). A second, slower time constant of 4-1 5 msec was found in the largest averaged EPSCs (greater-than-or-equal-to 40 pA). The amplitude of this slow component was -2 to -4 pA. These results suggest that, in the in vitro slice preparation, glutamate mediates all the spontaneous EPSCs in magnocellular neurosecretory cells by acting primarily on AMPA/kainate-type receptors at resting membrane potential and that activation of GABA(A) receptors mediates most if not all IPSCs.