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Journal of Neuroscience, Vol 13, 2323-2331, Copyright © 1993 by Society for Neuroscience
Patch-clamp analysis of spontaneous synaptic currents in supraoptic neuroendocrine cells of the rat hypothalamus
JP Wuarin and FE Dudek
Mental Retardation Research Center, UCLA School of Medicine 90024.
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 micrograms/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 microM and 2 cells at 20
microM) 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 GABAB 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 microM)
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
4-15 msec was found in the largest averaged EPSCs (> or = 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 GABAA receptors mediates most if not all IPSCs.
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