Cyclothiazide induces robust epileptiform activity in rat hippocampal neurons both in vitro and in vivo.
Cyclothiazide (CTZ) is a potent blocker of AMPA receptor desensitization. We have recently demonstrated that CTZ also inhibits GABA(A) receptors. Here we report that CTZ induces robust epileptiform activity in hippocampal neurons both in vitro and in vivo. We first found that chronic treatment of hippocampal cultures with CTZ (5 microM, 48 h) results in epileptiform activity in the majority of neurons (80%). The epileptiform activity lasts more than 48 h after washing off CTZ, suggesting a permanent change of the neural network properties after CTZ treatment. We then demonstrated in in vivo recordings that injection of CTZ (5 micromol in 5 microl) into the lateral ventricles of anaesthetized rats also induces spontaneous epileptiform activity in the hippocampal CA1 region. The epileptogenic effect of CTZ is probably due to its enhancing glutamatergic neurotransmission as shown by increasing the frequency and decay time of mEPSCs, and simultaneously inhibiting GABAergic neurotransmission by reducing the frequency of mIPSCs. Comparing to a well-known epileptogenic agent kainic acid (KA), CTZ affects neuronal activity mainly through modulating synaptic transmission without significant change of the intrinsic membrane excitability. Unlike KA, which induces significant cell death in hippocampal cultures, CTZ treatment does not result in any apparent neuronal death. Therefore, the CTZ-induced epilepsy model may provide a novel research tool to elucidate the molecular and cellular mechanisms of epileptogenesis without any complication from drug-induced cell death. The long-lasting epileptiform activity after CTZ washout may also make it a very useful model in screening antiepileptic drugs.
Qi
Jinshun
J
Department of Biology, 201 Life Sciences Building, The Pennsylvania State University, University Park, PA 16802, USA.
Wang
Yun
Y
Jiang
Min
M
Warren
Philippa
P
Chen
Gong
G
D000818:Animals; D000831:Animals, Newborn; D001581:Benzothiadiazines / Q000008:administration & dosage / Q000494:pharmacology*; D002118:Calcium / Q000378:metabolism; D002470:Cell Survival; D002478:Cells, Cultured; D004305:Dose-Response Relationship, Drug; D018690:Excitatory Amino Acid Agonists / Q000008:administration & dosage / Q000494:pharmacology* / Q000633:toxicity; D019706:Excitatory Postsynaptic Potentials; D006624:Hippocampus / Q000187:drug effects* / Q000502:physiology; D007276:Injections, Intraventricular; D007608:Kainic Acid / Q000633:toxicity; D009474:Neurons / Q000187:drug effects / Q000502:physiology; D051381:Rats; D017207:Rats, Sprague-Dawley
D001581:Benzothiadiazines; D018690:Excitatory Amino Acid Agonists; C004639:cyclothiazide; D007608:Kainic Acid; D002118:Calcium