Modulation of GABAA receptor-mediated inhibition by postsynaptic calcium in epileptic hippocampal neurons.
Visualization of neurons during patch clamp recordings from slices provides concurrent neuroanatomical information for physiological studies. Although, the technique becomes increasingly popular in immature brains, it has not been fully utilized in aged/adult and diseased brains including post-surgical human specimen. In the present study, glutamatergic modulation of GABAA receptor-mediated inhibition was investigated by whole-cell patch clamp recordings from visualized hippocampal dentate granule cells (DGCs) in slices that were prepared from surgically-removed human medial temporal lobe specimens and the rat pilocarpine model of temporal lobe epilepsy. GABAA receptor-mediated synaptic inhibition was recorded by isolating inhibitory postsynaptic currents (IPSCs) at a membrane potential of 0 mV where glutamatergic excitatory postsynaptic currents are near equilibrium. Peak amplitude of GABAA IPSC was not different between epileptic DGCs of both human and pilocarpine-treated rat hippocampi and those in the control rat DGCs. However, when high frequency stimulation (30 Hz for 10 s) preceded immediately before the generation of a GABAA IPSC, its peak amplitude was significantly reduced in epileptic DGCs. The application of an NMDA receptor antagonist prevented this decrease indicating that the high frequency stimulation activated the NMDA receptor and that this activation is involved in the induction of response-decrement of GABAA IPSCs in epileptic DGCs. In addition, intracellular application of a calcium chelator, BAPTA through a patch pipette was found effective in preventing the response-decrement of GABAA IPSCs suggesting that postsynaptic calcium-increase is also involved in this process. It is proposed that activation of the NMDA receptor in epileptic DGC may trigger an epileptogenic increase of intracellular free calcium, and this calcium-increase plays a crucial role for the induction of the response-decrement of GABAA IPSCs in epileptic hippocampus, which possibly leads to the initiation of epileptic seizures and ictal events.
Isokawa
M
M
Brain Research Institute, Center for Health Sciences, University of California, Los Angeles, CA 90095-1761, USA. misokawa@ucla.edu
D000818:Animals; D002118:Calcium / Q000502:physiology*; D002614:Chelating Agents / Q000494:pharmacology; D004558:Electric Stimulation; D004827:Epilepsy / Q000503:physiopathology*; D004833:Epilepsy, Temporal Lobe / Q000503:physiopathology; D006624:Hippocampus / Q000166:cytology / Q000503:physiopathology*; D006801:Humans; D066298:In Vitro Techniques; D008297:Male; D018721:Muscarinic Agonists / Q000494:pharmacology; D009474:Neurons / Q000187:drug effects / Q000502:physiology*; D018408:Patch-Clamp Techniques; D010862:Pilocarpine / Q000494:pharmacology; D051381:Rats; D017207:Rats, Sprague-Dawley; D011963:Receptors, GABA-A / Q000187:drug effects / Q000502:physiology*; D017981:Receptors, Neurotransmitter / Q000187:drug effects / Q000502:physiology*
D002614:Chelating Agents; D018721:Muscarinic Agonists; D011963:Receptors, GABA-A; D017981:Receptors, Neurotransmitter; D010862:Pilocarpine; D002118:Calcium