Neural Progenitor Cells Rptor Ablation Impairs Development but Benefits to Seizure-Induced Behavioral Abnormalities
Neural Progenitor Cells Rptor Ablation Impairs Development but Benefits to Seizure-Induced Behavioral Abnormalities
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DOI：
10.1111/cns.12607
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发表时间：
2016-12-01
影响因子：
5.5
通讯作者：
Zeng, Ling-Hui
中科院分区：
文献类型：
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作者：
Chen, Ling-Lin;Wu, Mei-Ling;Zeng, Ling-Hui
目的以往的研究表明mTOR信号通路可能在癫痫发生中起重要作用。目前的工作旨在探讨猛禽蛋白对癫痫和合并症发展的贡献。方法将raptor （flox/flox）小鼠与nesting - cre小鼠杂交，制备条件敲除raptor蛋白的smice。用Western blotting分析脑组织样品中raptor蛋白的表达。FJB染色和Timm染色分别检测神经元死亡和苔藓纤维发芽。自发性癫痫发作用脑电图视频系统记录。采用Morris水迷宫、开阔场实验和兴奋性实验研究Rptor CKO小鼠的行为。结果删除Rptor后，mTORC1信号通路中raptor下游蛋白被部分阻断。Rptor CKO小鼠在7周龄时体重和脑重下降，皮质层厚度下降。在kainic酸（KA）诱导的癫痫持续状态后，mTORC1信号的过度激活在Rptor CKO小鼠中被显著逆转。虽然在Rptor CKO和对照组中观察到自发性癫痫发作的频率较低，并且很少观察到神经元细胞死亡，但在Rptor CKO小鼠中，KA癫痫发作引起的苔藓纤维喷吐减弱。此外，在Rptor CKO小鼠中，ka诱发癫痫发作后的认知缺陷和焦虑样行为部分逆转。结论小鼠神经前体细胞中raptor基因缺失影响幼龄小鼠正常发育，并可能减轻KA癫痫引起的行为异常，提示raptor蛋白在癫痫合并症中起重要作用。
AimsPrevious study suggests that mTOR signaling pathway may play an important role in epileptogenesis. The present work was designed to explore the contribution of raptor protein to the development of epilepsy and comorbidities.MethodsMice with conditional knockout of raptor protein were generated by cross-bred Rptor(flox/flox) mice with nestin-CRE mice. The expression of raptor protein was analyzed by Western blotting in brain tissue samples. Neuronal death and mossy fiber sprouting were detected by FJB staining and Timm staining, respectively. Spontaneous seizures were recorded by EEG-video system. Morris water maze, open field test, and excitability test were used to study the behaviors of Rptor CKO mice.ResultsAs the consequence of deleting Rptor, downstream proteins of raptor in mTORC1 signaling were partly blocked. Rptor CKO mice exhibited decrease in body and brain weight under 7 weeks old and accordingly, cortical layer thickness. After kainic acid (KA)-induced status epilepticus, overactivation of mTORC1 signaling was markedly reversed in Rptor CKO mice. Although low frequency of spontaneous seizure and seldom neuronal cell death were observed in both Rptor CKO and control littermates, KA seizure-induced mossy fiber spouting were attenuated in Rptor CKO mice. Additionally, cognitive-deficit and anxiety-like behavior after KA-induced seizures were partly reversed in Rptor CKO mice.ConclusionLoss of the Rptor gene in mice neural progenitor cells affects normal development in young age and may contribute to alleviate KA seizure-induced behavioral abnormalities, suggesting that raptor protein plays an important role in seizure comorbidities.