PIP5K1α promotes myogenic differentiation via AKT activation and calcium release
Abstract Background Skeletal muscle satellite cell-derived myoblasts are mainly responsible for postnatal muscle growth and injury-induced regeneration. Many intracellular signaling pathways are essential for myogenic differentiation, while a number of kinases are involved in this modulation process...
| Main Authors: | , , , , |
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| Format: | Article |
| Language: | English |
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BMC
2018-02-01
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| Series: | Stem Cell Research & Therapy |
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| Online Access: | http://link.springer.com/article/10.1186/s13287-018-0770-z |
| _version_ | 1818140449670430720 |
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| author | Xiaofan Chen Jun Wan Bo Yu Yarui Diao Wei Zhang |
| author_facet | Xiaofan Chen Jun Wan Bo Yu Yarui Diao Wei Zhang |
| author_sort | Xiaofan Chen |
| collection | DOAJ |
| description | Abstract Background Skeletal muscle satellite cell-derived myoblasts are mainly responsible for postnatal muscle growth and injury-induced regeneration. Many intracellular signaling pathways are essential for myogenic differentiation, while a number of kinases are involved in this modulation process. Type I phosphatidylinositol 4-phosphate 5-kinase (PIP5KI) was identified as one of the key kinases involved in myogenic differentiation, but the underlying molecular mechanism is still unclear. Methods PIP5K1α was quantified by quantitative reverse transcriptase PCR and western blot assay. Expression levels of myogenin and myosin heavy chain, which showed significant downregulation in PIP5K1α siRNA-mediated knockdown cells in western blot analysis, were confirmed by immunostaining. Phosphatidylinositol 4,5-bisphosphate in PIP5K1α siRNA-mediated knockdown cells was also measured by the PI(4,5)P2 Mass ELISA Kit. C2C12 cells were overexpressed with different forms of AKT, followed by western blot analysis on myogenin and myosin heavy chain, which reveals their function in myogenic differentiation. FLIPR assays are used to test the release of calcium in PIP5K1α siRNA-mediated knockdown cells after histamine or bradykinin treatment. Statistical significances between groups were determined by two-tailed Student’s t test. Results Since PIP5K1α was the major form in skeletal muscle, knockdown of PIP5K1α consistently inhibited myogenic differentiation while overexpression of PIP5K1α promoted differentiation and rescued the inhibitory effect of the siRNA. PIP5K1α was found to be required for AKT activation and calcium release, both of which were important for skeletal muscle differentiation. Conclusions Taken together, these results suggest that PIP5K1α is an important regulator in myoblast differentiation. |
| first_indexed | 2024-12-11T10:44:10Z |
| format | Article |
| id | doaj.art-bab1997051d24ae1874058460914db80 |
| institution | Directory Open Access Journal |
| issn | 1757-6512 |
| language | English |
| last_indexed | 2024-12-11T10:44:10Z |
| publishDate | 2018-02-01 |
| publisher | BMC |
| record_format | Article |
| series | Stem Cell Research & Therapy |
| spelling | doaj.art-bab1997051d24ae1874058460914db802022-12-22T01:10:31ZengBMCStem Cell Research & Therapy1757-65122018-02-01911710.1186/s13287-018-0770-zPIP5K1α promotes myogenic differentiation via AKT activation and calcium releaseXiaofan Chen0Jun Wan1Bo Yu2Yarui Diao3Wei Zhang4Shenzhen Key Laboratory for Translational Medicine of Dermatology, Biomedical Research Institute, Shenzhen Peking University—the Hong Kong University of Science and Technology Medical CenterShenzhen Key Laboratory for Neuronal Structural Biology, Biomedical Research Institute, Shenzhen Peking University—the Hong Kong University of Science and Technology Medical CenterShenzhen Key Laboratory for Translational Medicine of Dermatology, Biomedical Research Institute, Shenzhen Peking University—the Hong Kong University of Science and Technology Medical CenterShenzhen Key Laboratory for Translational Medicine of Dermatology, Biomedical Research Institute, Shenzhen Peking University—the Hong Kong University of Science and Technology Medical CenterShenzhen Key Laboratory for Translational Medicine of Dermatology, Biomedical Research Institute, Shenzhen Peking University—the Hong Kong University of Science and Technology Medical CenterAbstract Background Skeletal muscle satellite cell-derived myoblasts are mainly responsible for postnatal muscle growth and injury-induced regeneration. Many intracellular signaling pathways are essential for myogenic differentiation, while a number of kinases are involved in this modulation process. Type I phosphatidylinositol 4-phosphate 5-kinase (PIP5KI) was identified as one of the key kinases involved in myogenic differentiation, but the underlying molecular mechanism is still unclear. Methods PIP5K1α was quantified by quantitative reverse transcriptase PCR and western blot assay. Expression levels of myogenin and myosin heavy chain, which showed significant downregulation in PIP5K1α siRNA-mediated knockdown cells in western blot analysis, were confirmed by immunostaining. Phosphatidylinositol 4,5-bisphosphate in PIP5K1α siRNA-mediated knockdown cells was also measured by the PI(4,5)P2 Mass ELISA Kit. C2C12 cells were overexpressed with different forms of AKT, followed by western blot analysis on myogenin and myosin heavy chain, which reveals their function in myogenic differentiation. FLIPR assays are used to test the release of calcium in PIP5K1α siRNA-mediated knockdown cells after histamine or bradykinin treatment. Statistical significances between groups were determined by two-tailed Student’s t test. Results Since PIP5K1α was the major form in skeletal muscle, knockdown of PIP5K1α consistently inhibited myogenic differentiation while overexpression of PIP5K1α promoted differentiation and rescued the inhibitory effect of the siRNA. PIP5K1α was found to be required for AKT activation and calcium release, both of which were important for skeletal muscle differentiation. Conclusions Taken together, these results suggest that PIP5K1α is an important regulator in myoblast differentiation.http://link.springer.com/article/10.1186/s13287-018-0770-zPIP5K1αMyogenic differentiationAKT activationCalcium release |
| spellingShingle | Xiaofan Chen Jun Wan Bo Yu Yarui Diao Wei Zhang PIP5K1α promotes myogenic differentiation via AKT activation and calcium release Stem Cell Research & Therapy PIP5K1α Myogenic differentiation AKT activation Calcium release |
| title | PIP5K1α promotes myogenic differentiation via AKT activation and calcium release |
| title_full | PIP5K1α promotes myogenic differentiation via AKT activation and calcium release |
| title_fullStr | PIP5K1α promotes myogenic differentiation via AKT activation and calcium release |
| title_full_unstemmed | PIP5K1α promotes myogenic differentiation via AKT activation and calcium release |
| title_short | PIP5K1α promotes myogenic differentiation via AKT activation and calcium release |
| title_sort | pip5k1α promotes myogenic differentiation via akt activation and calcium release |
| topic | PIP5K1α Myogenic differentiation AKT activation Calcium release |
| url | http://link.springer.com/article/10.1186/s13287-018-0770-z |
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