Differential molecular regulation of glutamate in kindling resistant rats
Differential molecular regulation of glutamate in kindling resistant rats
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DOI：
10.1016/j.brainres.2010.11.085
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发表时间：
2011-02-23
期刊：
影响因子：
2.9
通讯作者：
Willmore, L. James
中科院分区：
文献类型：
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作者：
Doi, Taku;Ueda, Yuto;Willmore, L. James
使用戊四氮（PTZ）点燃，我们收集海马组织从标准反应和点燃抗性动物，测量海马mRNA的实时PCR谷氨酸转运蛋白GLAST，GLT-1，和EAAC 1和钠偶联中性氨基酸转运蛋白（SNAT）1，SNAT 2，和SNAT 3。此外，我们还检测了谷氨酰胺合成酶（GS）、磷酸活化谷氨酰胺酶（PAG）、谷氨酸脱羧酶（GAD）1、GAD 2和囊泡抑制性氨基酸转运蛋白（VIAAT）的mRNA。与注射生理盐水的对照组相比，完全点燃的动物海马中GLAST和GAD 2的mRNA表达降低。SNAT 1、SNAT 2、SNAT 3、GS和VIAAT的mRNA增加。PTZ诱导全身强直阵挛性癫痫发作后，癫痫发作后24小时的mRNA无差异，除GAD 1外，其余均与基线水平相同。在PTZ诱导惊厥发作后30天测量水平时，我们发现GLT-1、SNAT 1和GS水平升高，但GAD 1水平降低。当这些动物作为点燃实验组中最后一次惊厥发作之间的30天间隔的对照时，我们发现GLT-1、SNAT 3、GAD 1和VIAAT的不同之处在于GLT-1减少而其他增加。发现对点燃有抵抗力的动物具有显著不同的mRNA模式，将谷氨酸转运到神经元和神经胶质的蛋白质的mRNA显著上调; SNAT 1也上调。抗点燃动物中基因的上调支持了谷氨酸的清除、谷氨酰胺的转化和谷氨酰胺转运到神经元中具有提高惊厥发作阈值和减弱点燃的作用的假设。(c)2010 Elsevier B. V.保留所有权利。
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 24 h after seizures, equaling baseline quantities except for GAD1, which was decreased. When levels were measured at 30 days 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 30 day 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. (c) 2010 Elsevier B.V. All rights reserved.