Direct inhibition of glycine receptors by genistein, a tyrosine kinase inhibitor

Genistein, a tyrosine kinase inhibitor, has been widely used to examine potential effects of protein tyrosine kinase (PTK)-mediated regulation of receptor/channel function. Alteration of ion channel function in the presence of genistein has typically led to the conclusion that PTK regulates the activity of the channel under investigation. In the present report, we have assessed the possibility that genistein directly inhibits the glycine receptor, independent of effects on protein tyrosine kinase. Coapplication of genistein with glycine reversibly inhibited the strychnine-sensitive, glycine-activated current recorded from hypothalamic neurons. The time course of genistein action was rapid (within ms). Equilibration of genistein in the intracellular solution did not affect the ability of extracellularly applied genistein to inhibit the glycine response. Glycine concentration–response profiles generated in the absence and presence of genistein indicated the block was due to non-competitive antagonism. The genistein effect also displayed voltage-dependence. Daidzein, an analog of genistein that does not block protein kinases, also inhibited glycine-activated current. Coapplication of lavendustin A, a specific inhibitor of PTK, had no effect on the glycine response. Our results demonstrate that the tyrosine kinase inhibitor genistein has a direct inhibitory effect on glycine receptors that is not mediated via inhibition of PTK.