Int J Biol Sci 2017; 13(1):110-121. doi:10.7150/ijbs.16312 This issue Cite

Research Paper

Involvement of Rictor/mTORC2 in cardiomyocyte differentiation of mouse embryonic stem cells in vitro

Bei Zheng1, Jiadan Wang1, Leilei Tang1, Chao Tan1, Zhe Zhao2, Yi Xiao2, Renshan Ge3, 4, Danyan Zhu1✉

1. Institute of Pharmacology and Toxicology, Zhejiang University, Hangzhou 310058, CHINA.
2. Undergraduate students in Research Training Project at Zhejiang University
Hangzhou 310058, CHINA.
3. The Population Council at the Rockefeller University, New York, NY 10021, USA.
4. Institute of Reproductive Biomedicine, the 2nd Affiliated Hospital of Wenzhou Medical University, Wenzhou 325027, CHINA.

Citation:
Zheng B, Wang J, Tang L, Tan C, Zhao Z, Xiao Y, Ge R, Zhu D. Involvement of Rictor/mTORC2 in cardiomyocyte differentiation of mouse embryonic stem cells in vitro. Int J Biol Sci 2017; 13(1):110-121. doi:10.7150/ijbs.16312. https://www.ijbs.com/v13p0110.htm
Other styles

File import instruction

Abstract

Graphic abstract

Rictor is a key regulatory/structural subunit of the mammalian target of rapamycin complex 2 (mTORC2) and is required for phosphorylation of Akt at serine 473. It plays an important role in cell survival, actin cytoskeleton organization and other processes in embryogenesis. However, the role of Rictor/mTORC2 in the embryonic cardiac differentiation has been uncovered. In the present study, we examined a possible link between Rictor expression and cardiomyocyte differentiation of the mouse embryonic stem (mES) cells. Knockdown of Rictor by shRNA significantly reduced the phosphorylation of Akt at serine 473 followed by a decrease in cardiomyocyte differentiation detected by beating embryoid bodies. The protein levels of brachyury (mesoderm protein), Nkx2.5 (cardiac progenitor cell protein) and α-Actinin (cardiomyocyte biomarker) decreased in Rictor knockdown group during cardiogenesis. Furthermore, knockdown of Rictor specifically inhibited the ventricular-like cells differentiation of mES cells with reduced level of ventricular-specific protein, MLC-2v. Meanwhile, patch-clamp analysis revealed that shRNA-Rictor significantly increased the number of cardiomyocytes with abnormal electrophysiology. In addition, the expressions and distribution patterns of cell-cell junction proteins (Cx43/Desmoplakin/N-cadherin) were also affected in shRNA-Rictor cardiomyocytes. Taken together, the results demonstrated that Rictor/mTORC2 might play an important role in the cardiomyocyte differentiation of mES cells. Knockdown of Rictor resulted in inhibiting ventricular-like myocytes differentiation and induced arrhythmias symptom, which was accompanied by interfering the expression and distribution patterns of cell-cell junction proteins. Rictor/mTORC2 might become a new target for regulating cardiomyocyte differentiation and a useful reference for application of the induced pluripotent stem cells.

Keywords: Rictor/mTORC2, embryonic stem cell, cardiomyocyte differentiation, cell-cell junction, electrophysiology.


Citation styles

APA
Zheng, B., Wang, J., Tang, L., Tan, C., Zhao, Z., Xiao, Y., Ge, R., Zhu, D. (2017). Involvement of Rictor/mTORC2 in cardiomyocyte differentiation of mouse embryonic stem cells in vitro. International Journal of Biological Sciences, 13(1), 110-121. https://doi.org/10.7150/ijbs.16312.

ACS
Zheng, B.; Wang, J.; Tang, L.; Tan, C.; Zhao, Z.; Xiao, Y.; Ge, R.; Zhu, D. Involvement of Rictor/mTORC2 in cardiomyocyte differentiation of mouse embryonic stem cells in vitro. Int. J. Biol. Sci. 2017, 13 (1), 110-121. DOI: 10.7150/ijbs.16312.

NLM
Zheng B, Wang J, Tang L, Tan C, Zhao Z, Xiao Y, Ge R, Zhu D. Involvement of Rictor/mTORC2 in cardiomyocyte differentiation of mouse embryonic stem cells in vitro. Int J Biol Sci 2017; 13(1):110-121. doi:10.7150/ijbs.16312. https://www.ijbs.com/v13p0110.htm

CSE
Zheng B, Wang J, Tang L, Tan C, Zhao Z, Xiao Y, Ge R, Zhu D. 2017. Involvement of Rictor/mTORC2 in cardiomyocyte differentiation of mouse embryonic stem cells in vitro. Int J Biol Sci. 13(1):110-121.

This is an open access article distributed under the terms of the Creative Commons Attribution (CC BY-NC) License. See http://ivyspring.com/terms for full terms and conditions.
Popup Image