Identification of RyR2-PBmice and the effects of transposon insertional mutagenesis of the RyR2 gene on cardiac function in mice
Identification of RyR2-PBmice and the effects of transposon insertional mutagenesis of the RyR2 gene on cardiac function in mice
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RyR2-PB小鼠的鉴定及RyR2基因转座子插入突变对小鼠心功能的影响
DOI：
10.7717/peerj.6942
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发表时间：
2019-05
期刊：
影响因子：
2.7
通讯作者：
Zhang Liming
中科院分区：
文献类型：
--
作者：
Wang Qianqian;Wang Chao;Wang Bo;Shen Qirui;Qiu Leilei;Zou Shuaijun;Wang Tao;Liu Guoyan;Wang Beilei;Zhang Liming
Ryanodine receptor 2（RyR 2）在维持正常心脏功能中起重要作用，其突变可导致心律失常、心力衰竭等心脏疾病。本研究采用红色荧光蛋白（RFP）追踪和基因分型PCR技术，成功地鉴定了一种piggyBac易位RyR 2基因杂合子小鼠模型（RyR 2-PB小鼠）。心功能测试显示，RyR 2-PB小鼠和相应的野生型小鼠（WT小鼠）之间没有显著差异，无论是在基础状态还是注射肾上腺素和咖啡因。然而，通过测量咖啡因诱导的[Ca 2 +]i瞬变来评估，RyR 2-PB小鼠心肌细胞中肌浆网Ca 2+含量显著降低; RyR 2-PB小鼠心肌组织显示出显著的线粒体肿胀和线粒体嵴局灶性溶解，并且RyR 2-PB小鼠心脏中的组织ATP含量显著降低。为了进一步分析这些变化背后的分子机制，我们使用RT-PCR和Western blot分析检测了相关蛋白的表达水平。与WT小鼠相比，RyR 2-PB小鼠心肌组织中RyR 2的mRNA水平显著降低，呼吸链相关蛋白的表达也下调。这些结果表明，插入RyR 2基因的piggyBac转座子实质上影响了小鼠心肌细胞线粒体的结构和功能，导致能量代谢障碍。
Ryanodine receptor 2 (RyR2) plays an important role in maintaining the normal heart function, and mutantions can lead to arrhythmia, heart failure and other heart diseases. In this study, we successfully identified a piggyBac translocated RyR2 gene heterozygous mouse model (RyR2-PBmice) by tracking red fluorescent protein (RFP) and genotyping PCR. Cardiac function tests showed that there was no significant difference between the RyR2-PBmice and corresponding wild-type mice (WTmice), regardless of whether they were in the basal state or injected with epinephrine and caffeine. However, the sarcoplasmic reticulum Ca2+ content was significantly reduced in the cardiomyocytes of RyR2-PBmice as assessed by measuring caffeine-induced [Ca2+]i transients; the cardiac muscle tissue of RyR2-PBmice displayed significant mitochondrial swelling and focal dissolution of mitochondrial cristae, and the tissue ATP content in the RyR2-PBmice heart was significantly reduced. To further analyze the molecular mechanism behind these changes, we tested the expression levels of related proteins using RT-PCR and Western blot analyses. The mRNA level of RyR2 in RyR2-PBmice cardiac tissue decreased significantly compared with the WTmice, and the protein expression associated with the respiratory chain was also downregulated. These results suggested that the piggyBac transposon inserted into the RyR2 gene substantively affected the structure and function of mitochondria in the mouse cardiomyocytes, leading to disorders of energy metabolism.
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影响因子：
20.1
作者：
K. Sipido
通讯作者：
K. Sipido
PBMICE：PIGGYBAC（PB）插入突变的集成数据库系统及其在小鼠中的表征。
影响因子：
14.9
作者：
Sun, Ling V.;Jin, Ke;Liu, Yiming;Yang, Wenwei;Xie, Xing;Ye, Lin;Wang, Li;Zhu, Lin;Ding, Sheng;Su, Yi;Zhou, Jie;Han, Min;Zhuang, Yuan;Xu, Tian;Wu, Xiaohui;Gu, Ning;Zhong, Yang
通讯作者：
Zhong, Yang
DOI：
10.1016/s0735-1097(02)01946-0
发表时间：
2002-07-17
影响因子：
24
作者：
Bauce, B;Rampazzo, A;Nava, A
通讯作者：
Nava, A
影响因子：
64.5
作者：
Wehrens, XHT;Lehnart, SE;Marks, AR
通讯作者：
Marks, AR
影响因子：
20.1
作者：
Cerrone, M;Colombi, B;Priori, SG
通讯作者：
Priori, SG