Fei Fei, Sun Shao-Yang, Yao Yu-Xiao, Wang Xu
Department of Biochemistry and Molecular Biology, School of Basic Medical Sciences; Key Laboratory of Metabolism and Molecular Medicine of Ministry of Education, Fudan University, Shanghai 200032, China.
Sheng Li Xue Bao. 2017 Feb 25;69(1):61-69.
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 mouse livers, and found that the changes in the expression of insulin/IGF signaling (IIS) pathway genes in lepr zebrafish and Lep 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.
肥胖已成为全球严重的公共卫生问题，严重影响人类健康和生活质量。在此，我们通过CRISPR/Cas9技术构建了lepr和mc4r突变斑马鱼，并对这些突变体进行了形态学和功能表征。我们观察到，在受精后2.5个月（mpf）之前，纯合突变体与野生型对照之间没有显著的表型差异。然而，成年lepr和mc4r个体表现出摄食量增加、体重增加和体脂百分比升高，这些都是肥胖表型的特征。血糖测试表明，过度喂养会导致成年lepr和mc4r斑马鱼的葡萄糖耐量显著受损。此外，我们通过实时RT-PCR分析了lepr和mc4r斑马鱼肝脏中76种与能量代谢相关的转录本，并将结果与已发表的Lep小鼠肝脏微阵列数据进行比较，发现lepr斑马鱼和Lep小鼠中胰岛素/IGF信号（IIS）通路基因表达的变化呈正相关，这表明在肥胖调节分子网络的进化过程中，IIS通路在斑马鱼和哺乳动物之间保持了功能保守性。
