Identification of RyR2-PBmice and the effects of transposon insertional mutagenesis of the RyR2 gene on cardiac function in mice

PeerJ

article

Qianqian Wang¹  Chao Wang¹  Bo Wang¹  Qirui Shen²  Leilei Qiu¹  Shuaijun Zou¹  Tao Wang³  Guoyan Liu¹  Beilei Wang¹  Liming Zhang¹ 

[1] Department of Marine Biotechnology, Faculty of Naval Medicine, Navy Medical University;School of Life Sciences, China Jiliang University;Department of Nuclear Medicine, Changhai Hospital, Navy Medical University

关键词: RyR2;    Piggybac;    Cardiomyocyte;    Mitochondrion;    Ca2+;   

DOI  :  10.7717/peerj.6942

学科分类：社会科学、人文和艺术（综合）

来源: Inra

【 摘 要 】

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.

【 授权许可】

CC BY