YTHDF1和YTHDF2在hSOD1^(G93A)突变型ALS转基因小鼠脊髓中的表达  

Expression of YTHDF1 and YTHDF2 in the spinal cord of mutant hSOD1^(G93A)ALS transgenic mice

作　　者：苏琦 管英俊 吕舒畅 陈晓素 陈燕春 张皓云 刘金梦 鲍炜炜 Su Qi;Guan Yingjun;Lv Shuchang;Chen Xiaosu;Chen Yanchun;Zhang Haoyun;Liu Jinmeng;Bao Weiwei(Histology and Embryology Department,Shandong Second Medical University,Weifang 261042,China;Neurologic Disorders and Regenerative Repair Lab of Shandong Higher Education,Weifang 261042,China)

机构地区：[1]山东第二医科大学组织学与胚胎学教研室,潍坊261042 [2]山东省高校神经疾病与再生修复实验室,潍坊261042

出　　处：《中国组织化学与细胞化学杂志》2024年第1期1-6,共6页Chinese Journal of Histochemistry and Cytochemistry

基　　金：国家自然科学基金(82271483);山东省自然科学基金(ZR2020MH150);山东省高等学校青创科技支持计划(2019KJK004)。

摘　　要：目的检测m^(6)A识别蛋白YTHDF1和YTHDF2在ALS转基因小鼠脊髓中的表达变化,阐明其异常表达与ALS疾病进展的动态关系。方法选取hSOD1^(G93A)突变型ALS转基因小鼠与同窝野生型(WT)小鼠,分别在发病前期(出生后70 d)、发病早期(出生后95 d)、发病中期(出生后108 d)和发病晚期(出生后122 d)分离脊髓,Western blot和免疫荧光染色双标技术检测YTHDF1和YTHDF2在小鼠脊髓中的蛋白表达与定位,qRT-PCR检测小鼠脊髓中YTHDF1和YTHDF2 mRNA表达。结果与WT小鼠相比,在70 d、95d、108 d和122 d ALS转基因小鼠脊髓中YTHDF1的mRNA水平和蛋白水平表达均降低,YTHDF2的mRNA水平和蛋白水平表达均升高。免疫荧光双标染色结果显示,在ALS转基因小鼠和WT小鼠脊髓前角均可检测到YTHDF1/神经元核抗原(NeuN)和YTHDF2/NeuN双阳性细胞,在疾病后期可检测到YTHDF1和YTHDF2与胶质纤维酸性蛋白(GFAP)双阳性细胞,提示YTHDF1和YTHDF2主要在脊髓神经元中表达。结论YTHDF1和YTHDF2在ALS转基因小鼠脊髓中表达异常与ALS脊髓运动神经元退变密切相关。Objective To detect the expression of m6A reader YTHDF1 and YTHDF2 in the spinal cord of amyotrophic lateral sclerosis(ALS)transgenic mice expressing mutant hSOD1^(G93A),elucidating the dynamic relationship between their abnormal ex-^(G93A)pression and the progression of ALS disease.Methods ALS transgenic mice expressing mutant hSOD1 and the same number of littermates wild-type mice were selected,and the spinal cords were isolated in the pre-symptomatic period(70th day after birth),early symptomatic(95th day after birth),mid-symptomatic(108th day after birth)and late symptomatic(122nd day after birth),and the pro-tein expression and localization of YTHDF1 and YTHDF2 in the spinal cord were detected by Western blot and immunofluorescence technique,and the mRNA expression of YTHDF1 and YTHDF2 in the spinal cord was detected by qRT-PCR.Results Compared to WT mice,in the spinal cords of ALS transgenic mice at 70,95,108 and 122 days,the mRNA and protein levels of YTHDF1 were decreased,while the mRNA and protein levels of YTHDF2 were increased.Immunofluorescence staining showed that YTHDF1/neu-ronal nuclear antigen(NeuN)and YTHDF2/NeuN double-positive cells were detectable in the anterior horn of the spinal cord of both ALS transgenic and WT mice.In the later stage of the disease,YTHDF1 and YTHDF2 double-positive cells with glial fibrillary acidic protein(GFAP)were detectable.It is indicated that YTHDF1 and YTHDF2 were mainly expressed in spinal neurons.Conclusion The abnormal expression of YTHDF1 and YTHDF2 in the spinal cord of ALS transgenic mice is closely related to degeneration of spinal motor neurons.

关 键 词：肌萎缩侧索硬化症 转基因小鼠 脊髓 m^(6)A修饰 YTHDF 

分 类 号：R744.8[医药卫生—神经病学与精神病学]