lepr和mc4r基因突变斑马鱼的制备及表型分析

发布时间：2018-06-28 02:01

本文选题：肥胖 + 斑马鱼　；参考：《生理学报》2017年01期

【摘要】：肥胖已经成为全球性的公共卫生问题,严重影响人类的身体健康和生活品质。本文运用CRISPR/Cas9技术,构建了肥胖相关基因——瘦素受体(leptin receptor,lepr)和黑素皮质素受体4(melanocortin-4 receptor,mc4r)的斑马鱼突变体,并对其进行形态和功能分析。结果显示,lepr~(-/-)和mc4r~(-/-)斑马鱼在胚胎和幼鱼期没有明显异常,但在成年期,lepr~(-/-)和mc4r~(-/-)斑马鱼相比对照进食增多、体重增加、体脂含量升高,呈现肥胖表型。血糖测定结果显示,饱食喂养条件可以诱导lepr~(-/-)和mc4r~(-/-)成年斑马鱼出现糖耐量受损的现象。进一步地,运用real time RT-PCR对76个能量代谢相关基因在lepr~(-/-)和mc4r~(-/-)斑马鱼肝脏中转录表达水平进行检测,并与Lep~(ob/ob)小鼠肝脏c DNA microarray的数据比较,发现胰岛素/胰岛素样生长因子信号转导通路(insulin/IGF signaling pathway,ISS)相关的基因在lepr~(-/-)斑马鱼和Lep~(ob/ob)小鼠肝脏中表达变化具有较高的正相关性,提示肥胖调控网络在进化中维持了一定的功能保守性。
[Abstract]:Obesity has become a global public health problem, seriously affecting human health and quality of life. Using CRISPRR / Cas9 technique, the zebrafish mutants of leptin receptor and melanocortin-4 receptor 4 were constructed, and their morphology and function were analyzed. The results showed that there was no obvious abnormality in embryonic and juvenile stages of zebrafish, but in adult and adult, zebrafish fed more, gained weight, increased body fat, and showed obesity phenotype. The results showed that full feeding could induce impaired glucose tolerance in adult zebrafish. Furthermore, real time RT-PCR was used to detect the transcriptional expression of 76 energy metabolism-related genes in the liver of lepr-r ~ (- / -) and mc4r-r ~ (- / -) zebrafish, and the results were compared with those of Lep~ (ob/ob) mouse liver. It was found that the expression of insulin / insulin-like growth factor (insulin / insulin-like growth factor signal transduction pathway) genes in the liver of lepr-r ~ (-) zebrafish and Lep~ (ob/ob) mice was positively correlated with the expression of insulin / insulin-like growth factor signal transduction pathway (insulin / insulin-like growth factor signal transduction pathway). These results suggest that the obesity regulatory network maintains a certain degree of functional conservatism in evolution.
【作者单位】：复旦大学基础医学院生物化学与分子生物学系代谢与分子医学教育部重点实验室;
【基金】：supported by grants from the National Natural Science Foundation of China(No.81402582) the Scientific Research Foundation for Returned Scholars,Ministry of Education of China(No.48-12)
【分类号】：R589.2

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