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Bicuculline is one of the most commonly used GABAAreceptor antagonists in electrophysiological research. Because of its poor water solubility, bicuculline quaternaryammonium salts such as bicuculline methiodide (BMI) andbicuculline methbromide are preferred. However, a numberof studies have shown that BMI has non-GABAA receptormediatedeffects. The substantia gelatinosa (SG) of thetrigeminal subnucleus caudalis (Vc) is implicated in theprocessing of nociceptive signaling. In this study, weinvestigated whether BMI has non-GABA receptormediatedactivity in Vc SG neurons using a whole cell patchclamp technique. SG neurons were depolarized byapplication of BMI (20 μM) using a high Cl-pipette solution. GABA (30-100 μM) also induced membrane depolarizationof SG neuron. Although BMI is known to be a GABAAreceptor antagonist, GABA-induced membrane depolarizationwas enhanced by co-application with BMI. However,free base bicuculline (fBIC) and picrotoxin (PTX), a GABAAand GABAC receptor antagonist, blocked the GABAinducedresponse. Furthermore, BMI-induced membranedepolarization persisted in the presence of PTX or anantagonist cocktail consisting of tetrodotoxin (Na+ channelblocker), AP-5 (NMDA receptor antagonist), CNQX (non-NMDA receptor antagonist), and strychnine (glycinereceptor antagonist). Thus BMI induces membranedepolarization by directly acting on postsynaptic Vc SGneurons in a manner which is independent of GABAAreceptors. These results suggest that other unknownmechanisms may be involved in BMI-induced membranedepolarization.

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