Söhl G, Güldenagel M, Beck H, Teubner B, Traub O, Gutiérrez R, Heinemann U, Willecke K
Abteilung für Molekulargenetik, Institut für Genetik, Universität Bonn, 53117, Bonn, Germany.
Brain Res Mol Brain Res. 2000 Nov 10;83(1-2):44-51. doi: 10.1016/s0169-328x(00)00195-9.
We have analyzed whether the expression of connexin genes is altered in the hippocampus of kindled and kainate-treated rats, i.e., animal models of human temporal lobe epilepsy. We have tested this hypothesis by analyzing mRNA, protein abundance and cellular location of connexins (Cx) 43, 36, 32 and 30. The expression of glial fibrillary acid protein and mRNA was also monitored both in kainate-treated and kindled rats, in order to take into account reactive gliosis under these conditions. We found significantly increased expression of GFAP mRNA (100%) and protein (178%) in kainate-treated rats 4 weeks after kainate application, whereas in kindled rats only moderate increases of GFAP mRNA and protein were detected 2-3 weeks (group 2) or 4-6 weeks (group 1) after the last stage 5 induced seizure. Under gliotic conditions, connexins 43 and 30 mRNA or protein expression in astrocytes of kainate-treated rats were nearly unaffected. Cx36 mRNA expression (presumably in neurons) was significantly reduced (44%), whereas abundance of Cx36 protein was only slightly reduced. In both groups of kindled rats, Cx30 and Cx43 mRNA or protein expression were either slightly decreased or unchanged. Again, Cx36 mRNA and protein expression were reduced by about half in group 2. Immunofluorescence analysis of Cx43, Cx36 and Cx30 expression revealed that 4 weeks after the last kainate administration or kindling, cellular localization of these connexins was indistinguishable from control animals.
我们分析了连接蛋白基因的表达在点燃模型大鼠和经海藻酸处理的大鼠(即人类颞叶癫痫的动物模型)海马体中是否发生改变。我们通过分析连接蛋白(Cx)43、36、32和30的mRNA、蛋白质丰度及细胞定位来验证这一假设。为了考虑在这些条件下的反应性胶质增生,我们还监测了经海藻酸处理的大鼠和点燃模型大鼠中胶质纤维酸性蛋白及其mRNA的表达。我们发现,在应用海藻酸4周后的经海藻酸处理的大鼠中,GFAP mRNA(增加100%)和蛋白质(增加178%)的表达显著增加,而在点燃模型大鼠中,在最后一次5期诱发癫痫发作后的2 - 3周(第2组)或4 - 6周(第1组),仅检测到GFAP mRNA和蛋白质有适度增加。在胶质增生的条件下,经海藻酸处理的大鼠星形胶质细胞中连接蛋白43和30的mRNA或蛋白质表达几乎未受影响。Cx36 mRNA表达(可能在神经元中)显著降低(44%),而Cx36蛋白质的丰度仅略有降低。在两组点燃模型大鼠中,Cx30和Cx43的mRNA或蛋白质表达要么略有下降,要么没有变化。同样,第2组中Cx36的mRNA和蛋白质表达减少了约一半。对Cx43、Cx36和Cx30表达的免疫荧光分析显示,在最后一次给予海藻酸或点燃后的4周,这些连接蛋白的细胞定位与对照动物没有区别。