Kashihara K, Sato K, Akiyama K, Okada S, Ishihara T, Hayabara T, Shomori T
Department of Neurology, Okayama University Medical School, Japan.
Neuroscience. 1997 Oct;80(3):753-61. doi: 10.1016/s0306-4522(97)00133-4.
DNA binding by transcripton factor AP-1 was enhanced remarkably following kindling stimulation in rat amygdala. Maximum increase occurred 2 h after stimulation with return to baseline within 24 h. Supershift and western analyses revealed that 38,000 mol. wt Fos-related antigen and JunD were the main components of the evoked AP-1 complexes at the time their induction reached maximum. AP-1 induction 2 h after the last kindling stimulation was more prominent in samples from previously kindled rats than in those from non-kindled rats. This study sought to establish the role of AP-1 in plastic changes of the hippocampus associated with kindling. Male Sprague-Dawley rats were kindled from the left amygdala until they exhibited Racine15 class 5 generalized seizures. Nuclear proteins were extracted from dorsal hippocampi obtained from 0 to 24 h after final stimulations. From these, we evaluated the temporal pattern of DNA binding by AP-1 using a gel mobility-shift assay with a 32P-labelled AP-1 probe. Supershift and western analyses were added to investigate components of the seizure-evoked AP-1 complexes. Our results suggest that the basal level of AP-1 complexes is not associated with the seizure susceptibility in kindling. However, development of kindling appears to facilitate stimulus-evoked AP-1 induction, probably via plastic changes in the central nervous system. AP-1 may mediate such changes by regulating expression of certain genes.
在大鼠杏仁核点燃刺激后,转录因子AP-1与DNA的结合显著增强。刺激后2小时出现最大增幅,并在24小时内恢复至基线水平。超迁移和蛋白质印迹分析显示,在诱导达到最大值时,分子量为38,000的Fos相关抗原和JunD是诱发的AP-1复合物的主要成分。末次点燃刺激后2小时,先前点燃大鼠样本中的AP-1诱导比未点燃大鼠样本中更明显。本研究旨在确定AP-1在与点燃相关的海马体可塑性变化中的作用。雄性Sprague-Dawley大鼠从左侧杏仁核开始点燃,直至出现Racine 15级5型全身性惊厥。在末次刺激后0至24小时从背侧海马体中提取核蛋白。从中,我们使用32P标记的AP-1探针通过凝胶迁移率变动分析评估AP-1与DNA结合的时间模式。添加超迁移和蛋白质印迹分析以研究癫痫发作诱发的AP-1复合物的成分。我们的结果表明,AP-1复合物的基础水平与点燃中的癫痫易感性无关。然而,点燃的发展似乎促进了刺激诱发的AP-1诱导,可能是通过中枢神经系统的可塑性变化。AP-1可能通过调节某些基因的表达来介导这种变化。