Generation and phenotype analysis of zebrafish mutations of obesity-related genes lepr and mc4r
Generation and phenotype analysis of zebrafish mutations of obesity-related genes lepr and mc4r
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DOI：
10.13294/j.aps.2016.0076
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发表时间：
2017-02-25
期刊：
影响因子：
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通讯作者：
Wang Xu
中科院分区：
文献类型：
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作者：
Fei Fei;Sun Shao-Yang;Wang Xu
肥胖已成为一个严重的全球性公共卫生问题，严重影响人类的健康和生活质量。在这里，我们通过CRISPR/Cas9技术产生了lepr和mc 4 r突变斑马鱼，并对这些突变体进行了形态和功能表征。我们观察到，在受精后2.5个月（mpf）之前，纯合子突变体和野生型对照之间没有显著的表型差异。然而，成年lepr（-/-）和mc 4 r（-/-）个体表现出食物摄入量增加、体重增加和体脂百分比升高等肥胖表型特征。血糖测定结果表明，过量投喂可显著降低lepr（-/-）和mc 4 r（-/-）斑马鱼的糖耐量。此外，我们利用实时荧光定量RT-PCR技术分析了lepr（-/-）和mc 4 r（-/-）斑马鱼肝脏中76个能量代谢相关转录本，并将结果与已发表的Lep（ob/ob）小鼠肝脏基因芯片数据进行了比较，发现lepr（-/-）斑马鱼和Lep（ob/ob）小鼠胰岛素/IGF信号通路（IIS）基因的表达变化呈正相关，这表明IIS途径在肥胖调节分子网络的进化过程中保持了斑马鱼和哺乳动物之间的功能保守性。
Obesity has become a severe public health problem across the world, and seriously affects the health and life quality of human beings. Here we generated lepr and mc4r mutant zebrafish via the CRISPR/Cas9 technique, and performed morphological and functional characterizations of those mutants. We observed that there was no significant phenotypic difference between homozygous mutants and wild-type controls before 2.5 months post-fertilization (mpf). However, the adult lepr(-/-) and mc4r(-/-) individuals displayed increased food intake, heavier weight, and higher body fat percentage, the characteristics of obesity phenotypes. Blood glucose test showed that overfeeding induced significantly impaired glucose tolerance in adult lepr(-/-) and mc4r(-/-) zebrafish. Furthermore, we analyzed 76 energy metabolism-related transcripts in lepr(-/-) and mc4r(-/-) zebrafish livers by using real-time RT-PCR, and compared the results with the published microarray data of Lep(ob/ob) mouse livers, and found that the changes in the expression of insulin/IGF signaling (IIS) pathway genes in lepr(-/-) zebrafish and Lep(ob/ob) mouse were positively correlated, suggesting that the IIS pathway maintains functional conservation between zebrafish and mammals during the evolution of the obesity-regulating molecule network.