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Overexpression of Cardiac-Specific Kinase TNNI3K Promotes Mouse Embryonic Stem Cells Differentiation into Cardiomyocytes
- Source :
- Cellular Physiology and Biochemistry, Vol 41, Iss 1, Pp 381-398 (2017)
- Publication Year :
- 2017
- Publisher :
- Cell Physiol Biochem Press GmbH & Co KG, 2017.
-
Abstract
- Backgroud/Aims: The biological function of cardiac troponin I-interacting kinase (TNNI3K), a cardiac-specific functional kinase, is largely unknown. We investigated the effect of human TNNI3K (hTNNI3K) on the differentiation of mouse embryonic stem cells (mESCs) into cardiomyocytes. Methods: First, the time-space expression of endogenous Tnni3k was detected by real-time polymerase chain reaction (PCR) and western blotting at 16 different time-points over a period of 28 days. Further, action potentials and calcium current with/without 5 µM nifedipine were measured by patch clamp for mESC-derived cardiomyocytes. HTNNI3K and mouse-derived siRNA were transfected into mESC using lentivirus vector to induce hTNNI3K overexpression and knock-down, respectively. Results: The number of troponin-T (cTnT) positive cells was greater in the group with TNNI3K overexpression as compared to that in control group, while less such cells were detected in the mTnni3k knock-down group as evaluated on flow cytometry (FCM) and ImageXpress Micro system. After upregulation of connexin43, cardiac troponin-I (Ctni), Ctni, Gata4 were detected in mESCs with TNNI3K overexpression; however, overexpression of α-Actinin and Mlc2v was not detected. Interestingly, Ctnt, connexin40 and connexin45, the markers of ventricular, atrial, and pacemaker cells, respectively, were detected in by real-time PCR in TNNI3K overexpression group. Conclusion: our study indicated that TNNI3K overexpression promoted mESC differentiating into beating cardiomyocytes and induced up-regulating expression of cTnT by PKCε signal pathway, which suggested a modulation of TNNI3K activity as a potential therapeutic approach for ischemic cardiac disease.
- Subjects :
- TNNI3K
mESC
Differentiation
Cardiomyocyte
Physiology
QP1-981
Biochemistry
QD415-436
Subjects
Details
- Language :
- English
- ISSN :
- 10158987, 14219778, and 00045640
- Volume :
- 41
- Issue :
- 1
- Database :
- Directory of Open Access Journals
- Journal :
- Cellular Physiology and Biochemistry
- Publication Type :
- Academic Journal
- Accession number :
- edsdoj.98646999b33340bd929d2c4ef8cec341
- Document Type :
- article
- Full Text :
- https://doi.org/10.1159/000456400