Identification of RyR2-PBmice and the effects of transposon insertional mutagenesis of the RyR2 gene on cardiac function in mice
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...
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PeerJ Inc.
2019-05-01
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| author | Qianqian Wang Chao Wang Bo Wang Qirui Shen Leilei Qiu Shuaijun Zou Tao Wang Guoyan Liu Beilei Wang Liming Zhang |
| author_facet | Qianqian Wang Chao Wang Bo Wang Qirui Shen Leilei Qiu Shuaijun Zou Tao Wang Guoyan Liu Beilei Wang Liming Zhang |
| author_sort | Qianqian Wang |
| collection | DOAJ |
| description | 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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| language | English |
| last_indexed | 2024-03-09T07:23:34Z |
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| spelling | doaj.art-090a3382881a40268dc698eef250938d2023-12-03T07:09:15ZengPeerJ Inc.PeerJ2167-83592019-05-017e694210.7717/peerj.6942Identification of RyR2-PBmice and the effects of transposon insertional mutagenesis of the RyR2 gene on cardiac function in miceQianqian Wang0Chao Wang1Bo Wang2Qirui Shen3Leilei Qiu4Shuaijun Zou5Tao Wang6Guoyan Liu7Beilei Wang8Liming Zhang9Department of Marine Biotechnology, Faculty of Naval Medicine, Navy Medical University, Shanghai, ChinaDepartment of Marine Biotechnology, Faculty of Naval Medicine, Navy Medical University, Shanghai, ChinaDepartment of Marine Biotechnology, Faculty of Naval Medicine, Navy Medical University, Shanghai, ChinaSchool of Life Sciences, China Jiliang University, Hangzhou, ChinaDepartment of Marine Biotechnology, Faculty of Naval Medicine, Navy Medical University, Shanghai, ChinaDepartment of Marine Biotechnology, Faculty of Naval Medicine, Navy Medical University, Shanghai, ChinaDepartment of Nuclear Medicine, Changhai Hospital, Navy Medical University, Shanghai, ChinaDepartment of Marine Biotechnology, Faculty of Naval Medicine, Navy Medical University, Shanghai, ChinaDepartment of Marine Biotechnology, Faculty of Naval Medicine, Navy Medical University, Shanghai, ChinaDepartment of Marine Biotechnology, Faculty of Naval Medicine, Navy Medical University, Shanghai, ChinaRyanodine 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.https://peerj.com/articles/6942.pdfRyR2PiggybacCardiomyocyteMitochondrionCa2+ |
| spellingShingle | Qianqian Wang Chao Wang Bo Wang Qirui Shen Leilei Qiu Shuaijun Zou Tao Wang Guoyan Liu Beilei Wang Liming Zhang Identification of RyR2-PBmice and the effects of transposon insertional mutagenesis of the RyR2 gene on cardiac function in mice PeerJ RyR2 Piggybac Cardiomyocyte Mitochondrion Ca2+ |
| title | Identification of RyR2-PBmice and the effects of transposon insertional mutagenesis of the RyR2 gene on cardiac function in mice |
| title_full | Identification of RyR2-PBmice and the effects of transposon insertional mutagenesis of the RyR2 gene on cardiac function in mice |
| title_fullStr | Identification of RyR2-PBmice and the effects of transposon insertional mutagenesis of the RyR2 gene on cardiac function in mice |
| title_full_unstemmed | Identification of RyR2-PBmice and the effects of transposon insertional mutagenesis of the RyR2 gene on cardiac function in mice |
| title_short | Identification of RyR2-PBmice and the effects of transposon insertional mutagenesis of the RyR2 gene on cardiac function in mice |
| title_sort | identification of ryr2 pbmice and the effects of transposon insertional mutagenesis of the ryr2 gene on cardiac function in mice |
| topic | RyR2 Piggybac Cardiomyocyte Mitochondrion Ca2+ |
| url | https://peerj.com/articles/6942.pdf |
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