YTHDF1和YTHDF2在hSOD1G93A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 hSOD1G93A ALS transgenic mice
苏琦 1管英俊 2吕舒畅 1陈晓素 1陈燕春 1张皓云 3刘金梦 3鲍炜炜1
作者信息
- 1. 山东第二医科大学组织学与胚胎学教研室,潍坊 261042;山东省高校神经疾病与再生修复实验室,潍坊 261042
- 2. 山东第二医科大学组织学与胚胎学教研室,潍坊 261042
- 3. 山东省高校神经疾病与再生修复实验室,潍坊 261042
- 折叠
摘要
目的 检测m6A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hSOD1G93A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.
Abstract
Objective To detect the expression of m6A reader YTHDF1 and YTHDF2 in the spinal cord of amyotrophic lateral sclerosis(ALS)transgenic mice expressing mutant hSOD1G93A,elucidating the dynamic relationship between their abnormal ex-pression and the progression of ALS disease.Methods ALS transgenic mice expressing mutant hSOD1G93A 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/m6A修饰/YTHDFKey words
Amyotrophic lateral sclerosis/transgenic mice/spinal cord/N6-methyladenosine(m6A)/YTHDF引用本文复制引用
基金项目
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)
出版年
2024
NETL
NSTL
万方数据