Volume 17, Number 3,
Issue of February 1, 1997
pp. 941-950
Copyright ©1997 Society for Neuroscience
N-Ethylmaleimide Blocks Depolarization-Induced
Suppression of Inhibition and Enhances GABA Release in the Rat
Hippocampal Slice In Vitro
Received Aug. 12, 1996; revised Nov. 5, 1996; accepted Nov. 7, 1996.
Wade Morishita,
Sergei A. Kirov,
Thomas A. Pitler,
Laura
A. Martin,
Robert A. Lenz, and
Bradley E. Alger
Department of Physiology, University of Maryland School of
Medicine, Baltimore, Maryland 21201
Regulation of synaptic, GABAA receptor-mediated
inhibition is a process of critical importance to normal brain
function. Recently, we have described a phenomenon in hippocampus of a
transient, yet marked, decrease in spontaneous, GABAA
receptor-mediated IPSCs after depolarization activated Ca2+
influx into a pyramidal cell. This process, depolarization-induced suppression of inhibition (DSI), is absent in hippocampal cells that
previously had been exposed to pertussis toxin in vivo,
implicating a G-protein in the DSI process. To circumvent the problem
that a single cell cannot be studied before and after G-protein block using the pertussis toxin pretreatment method, we have used the sulfhydryl alkylating agent N-ethylmaleimide (NEM),
which blocks pertussis toxin-sensitive G-proteins, to determine whether
acute inhibition of G-proteins can eliminate DSI of spontaneous IPSCs (sIPSCs). In whole-cell recordings from CA1 pyramidal cells that were
first determined to express DSI, we have found that NEM does block DSI
of sIPSCs. We also report that DSI of monosynaptic, evoked IPSCs is
blocked by NEM, suggesting that a similar mechanism underlies both
forms of DSI. It was of interest that DSI was abolished at a time when
NEM had increased, not decreased, GABA transmission. Indeed, NEM
greatly increased quantal GABA release by a
Ca2+-independent mechanism, an observation with potentially
important implications for understanding synaptic GABA release.
Key words:
NEM;
hippocampus;
GABA;
mIPSCs;
transmitter release;
IPSC