Differential molecular regulation of glutamate in kindling resistant rats.
Using pentylenetetrazol (PTZ) kindling, we collected hippocampal tissue from standard response and kindling resistant animals, measuring hippocampal mRNA with real-time PCR of glutamate transporters GLAST, GLT-1, and EAAC1 and the sodium-coupled neutral amino acid transporter (SNAT) 1, SNAT2, and SNAT3. In addition, we measured mRNA of glutamine synthetase (GS), phosphate-activated glutaminase (PAG), glutamic acid decarboxylase (GAD) 1, GAD2, and vesicular inhibitory amino acid transporter (VIAAT). Fully kindled animals had decreased expression of mRNA in the hippocampus for GLAST and GAD2 compared with saline injected control. mRNA for SNAT1, SNAT2, SNAT3, GS, and VIAAT was increased. After induction of generalized tonic-clonic seizures by PTZ there were no differences in mRNA at 24h after seizures, equaling baseline quantities except for GAD1, which was decreased. When levels were measured at 30days after a PTZ induced convulsive seizure, we found increased levels of GLT-1, SNAT1 and GS, but decreased levels of GAD1. When these animals, serving as control for the 30day interval between the last convulsive seizure in the kindled experimental group, were analyzed, we found that GLT-1, SNAT3, GAD1 and VIAAT differed in that GLT-1 was decreased and the others increased. Animals found resistant to kindling had strikingly different mRNA patterns, with markedly up-regulated mRNA of proteins that transport glutamate into neurons and glia; SNAT1 was up regulated as well. Up-regulation of genes in kindling resistant animals supports the hypothesis that clearance of glutamate, conversion to glutamine and transport of glutamine into neurons, has the effect of raising the threshold for convulsive seizures and attenuating kindling.
Section of Psychiatry, Department of Clinical Neuroscience, Faculty of Medicine, University of Miyazaki, 5200 Kihara, Kiyotake, Miyazaki, 889-1692, Japan. takud@med.miyazaki-u.ac.jp
Taku
T
Doi
Yuto
Y
Ueda
Mayuko
M
Takaki
L James
LJ
Willmore
D000818:Animals; D003292:Convulsants; D017931:DNA Primers; D004558:Electric Stimulation; D004830:Epilepsy, Tonic-Clonic / Q000139:chemically induced / Q000378:metabolism; D018698:Glutamic Acid / Q000378:metabolism* / Q000502:physiology; D006624:Hippocampus / Q000378:metabolism / Q000502:physiology; D007696:Kindling, Neurologic / Q000502:physiology*; D008297:Male; D010433:Pentylenetetrazole; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D051381:Rats; D017207:Rats, Sprague-Dawley; D020133:Reverse Transcriptase Polymerase Chain Reaction; D048348:Reverse Transcription; D012640:Seizures / Q000139:chemically induced / Q000503:physiopathology; D013268:Stimulation, Chemical
D003292:Convulsants; D017931:DNA Primers; D012333:RNA, Messenger; D018698:Glutamic Acid; D010433:Pentylenetetrazole