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作 者:蒋欣 管英俊 陈燕春[1] 刘金梦 周风华[1] 王巧真[1] 张皓云[1] Jiang Xin;Guan Yingjun;Chen Yanchun;Liu Jinmeng;Zhou Fenghua;Wang Qiaozhen;Zhang Haoyun(Clinical Medical College,Weifang Medical University,Weifang 261053,China;Laboratory of Neurologic disorders and regenerative repair,Weifang Medical University,Weifang 261053,China)
机构地区:[1]潍坊医学院临床医学院,潍坊261053 [2]潍坊医学院神经疾病与再生修复实验室,潍坊261053
出 处:《中国组织化学与细胞化学杂志》2019年第4期318-323,共6页Chinese Journal of Histochemistry and Cytochemistry
基 金:国家自然科学基金(81871006);山东省高等学校科研发展计划项目重点项目(J18KZ013)
摘 要:目的探讨海马CaMKⅡ-α和CaMKⅡ-β表达变化与肌萎缩侧索硬化症(amyotrophic lateral sclerosis,ALS)病变的关系。方法运用ALS转基因小鼠,采用免疫荧光技术检测海马中CaMKⅡ-α、CaMKⅡ-β的表达情况和细胞类型;应用RT-PCR和Western blot技术检测CaMKⅡ-α、CaMKⅡ-βmRNA和蛋白的变化。结果免疫荧光染色技术显示ALS转基因小鼠海马的CaMKⅡ-α免疫反应性明显降低,CaMKⅡ-β则明显升高;CaMKⅡ-α和CaMKⅡ-β阳性细胞均与β-tubulinⅢ标记的神经元共存;RT-PCR结果显示CaMKⅡ-α、CaMKⅡ-βmRNA水平无显著性变化;Western blot结果显示CAMKⅡ-α蛋白表达降低,CAMKⅡ-β蛋白表达升高。结论海马神经元CaMKⅡ-α蛋白降低和CaMKⅡ-β蛋白升高与ALS病变相关。Objective To explore the relationship between the expression of CaMKⅡ-αand CaMKⅡ-βin the hippocampus and the pathogenesis of amyotrophic lateral sclerosis(ALS).Methods The expression and cell type of CaMKⅡ-αand CaMKⅡ-βin the hippocampus of ALS transgenic mice were detected by immunofluorescence technique,and the expression changes of CaMKⅡ-αand CaMKⅡ-βmRNA and protein in the hippocampus of ALS transgenic mice were detected by RT-PCR and Western blot.Results Immu-nofluorescence staining showed that CaMKⅡ-αimmunoreactivity of ALS transgenic mice was lower than that of WT mice,while the immunoreactivity of CaMKⅡ-βwas increased.Both CaMKⅡ-αpositive and CaMKⅡ-βpositive cells were co-localized withβ-tubulinⅢlabeled neurons in the hippocampus of ALS transgenic mice and WT mice.RT-PCR results showed that there was no obvious dif-ference in the levels of CaMKⅡ-αand CaMKⅡ-βmRNA.Western bolt analysis demonstrated that the expression of CaMKⅡ-αprotein was decreased and the expression of CaMKⅡ-βprotein was increased,compared with WT mice.Conclusion Decreased expression of CaMKⅡ-αprotein and increased expression of CaMKⅡ-βprotein in hippocampal neurons may be related to the pathogenesis of ALS.
关 键 词:萎缩侧索硬化症 CaMKⅡ-α CaMKⅡ-β 海马 转基因小鼠
分 类 号:R741.02[医药卫生—神经病学与精神病学]
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