Myh13敲除小鼠表型的初步分析

doi:10.3969/j.issn.1674-5817.2019.03.004

摘要/Abstract

摘要： 目的对Myh13基因敲除小鼠模型进行初步的表型分析,探讨Myh13的生物学功能。方法利用qRT-PCR和Western blotting技术检测小鼠中Myh13的表达情况; 采用基因测序、qRT-PCR和Western blotting法验证Myh13敲除小鼠; 统计杂合子小鼠自交产生的子代小鼠中各基因型比例; 观察小鼠体质量变化趋势,并检测血液生化指标; 对小鼠眼外肌进行HE染色和透射电子显微镜观察。结果在小鼠模型中,Myh13特异性高表达于眼外肌组织。经DNA、RNA和蛋白水平验证,Myh13敲除小鼠中目的基因成功剔除。杂合子小鼠子代中各基因型小鼠比例符合孟德尔遗传规律。Myh13敲除对小鼠的体质量、血生化指标无明显影响。HE染色结果提示,与野生型小鼠相比,杂合子和纯合子小鼠的眼外肌均无明显异常。电子显微镜下,杂合子和纯合子小鼠眼外肌中可见散在脂滴,并且纯合子小鼠还可见肌质网扩张的现象。结论 Myh13的敲除可改变小鼠眼外肌的微观结构,可能会影响眼外肌的功能。

关键词: Myh13, 小鼠模型, 眼外肌

Abstract: Objective A preliminary phenotypic analysis of the Myh13 knockout mouse model was performed in order to investigate the biological function of Myh13. Methods Expression of Myh13 in mice was determined by qRT-PCR and Western blotting. Validation of Myh13 knockout mouse was analyzed by gene sequencing, qRT-PCR and Western blotting. Genotypic ratio in the offsprings from the self-cross of heterozygotes was calculated. Changes in body weight and serum biochemistry parameters were measured. Histopathological features of mouse extraocular muscles were displayed by H&E staining and transmission electron microscopy examination. Results In mice, Myh13 is specifically expressed in extraocular muscle. It was verified that the target gene in Myh13 knockout mouse was successfully deleted at DNA, RNA and protein levels. Genotypic ratio of heterozygotes in the offsprings was consistent with Mendel’s law. No significant differences existed among different genotypes in body weight or serum biochemical analysis. HE staining revealed no remarkable change in homozygous and heterozygous mouse extraocular muscles compared to wild-type mouse extraocular muscles. Electron microscopy of mouse extraocular muscles showed that fat droplets deposited in heterozygotes and homozygotes. Sarcoplasmic reticulum dilation was also identified in homozygotes. Conclusions Knockout of Myh13 can alter the microstructure of extraocular muscles in mice, which may affect the function of extraocular muscles.

Key words: Myh13, Mouse model, Extraocular muscle

中图分类号:

Q95-33

何一旻, 顾鸣敏. Myh13敲除小鼠表型的初步分析[J]. 实验动物与比较医学, 2019, 39(3): 193-200.

HE Yi-min, GU Ming-min. Preliminary Phenotypic Analysis of Myh13 Knockout Mouse[J]. Laboratory Animal and Comparative Medicine, 2019, 39(3): 193-200.

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