Inhibition of calcium/calmodulin kinase II alpha subunit expression results in epileptiform activity in cultured hippocampal neurons
Inhibition of calcium/calmodulin kinase II alpha subunit expression results in epileptiform activity in cultured hippocampal neurons
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DOI：
10.1073/pnas.080071697
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发表时间：
2000-05-09
影响因子：
11.1
通讯作者：
DeLorenzo, RJ
中科院分区：
文献类型：
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作者：
Churn, SB;Sombati, S;DeLorenzo, RJ
发展癫痫样放电和癫痫的几种模型与钙调蛋白依赖性激酶II活性的降低有关。然而，这些研究均未证明钙/钙调蛋白激酶II活性降低与癫痫发作活动发展之间存在因果关系。本研究旨在确定直接降低钙/钙调素依赖性激酶活性对培养海马神经元癫痫样放电发展的影响。特异于钙/钙调蛋白激酶α亚基的互补寡核苷酸用于降低酶的表达。通过Western分析、免疫细胞化学和外源性底物磷酸化证实激酶表达减少。增加神经元兴奋性和坦率的癫痫样放电后，观察到钙调蛋白激酶II的表达显着减少。癫痫样活动是一个同步事件，而不是由随机神经元放电。此外，激酶表达降低的幅度与神经元兴奋性增加相关。这些数据表明，钙调蛋白激酶II活性降低可能在癫痫发生和长期可塑性变化与病理性癫痫发作活动和癫痫的发展中发挥作用。
Several models that develop epileptiform discharges and epilepsy have been associated with a decrease in the activity of calmodulin-dependent kinase II. However, none of these studies has demonstrated a causal relationship between a decrease in calcium/calmodulin kinase II activity and the development of seizure activity. The present study was conducted to determine the effect of directly reducing calcium/calmodulin-dependent kinase activity on the development of epileptiform discharges in hippocampal neurons in culture. Complimentary oligonucleotides specific for the alpha subunit of the calciumlcalmodulin kinase were used to decrease the expression of the enzyme. Reduction in kinase expression was confirmed by Western analysis, immunocytochemistry, and exogenous substrate phosphorylation. Increased neuronal excitability and frank epileptiform discharges were observed after a significant reduction in calmodulin kinase II expression. The epileptiform activity was a synchronous event and was not caused by random neuronal firing. Furthermore, the magnitude of decreased kinase expression correlated with the increased neuronal excitability. The data suggest that decreased calmodulin kinase II activity may play a role in epileptogenesis and the long-term plasticity changes associated with the development of pathological seizure activity and epilepsy.