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Published on: May 17, 2016
Intracellular TMEM16A is necessary for myogenesis of skeletal muscle
Wen Yuan1, Cong-Cong Cui1, Jing Li1
1The Jiangsu Key Laboratory for Molecular and Medical Biotechnology, School of Life Sciences, Nanjing Normal University, Nanjing 210023, China.
Abstract:
Transmembrane protein 16A (TMEM16A) localizes at plasma membrane and controls chloride influx in various type of cells. We here showed an intracellular localization pattern of TMEM16A molecules. In myoblasts, TMEM16A was primarily localized to the cytosolic compartment and partially co-localized with intracellular organelles. The global deletion of TMEM16A led to severe skeletal muscle developmental defect. In vitro observation showed that the proliferation of Tmem16a-/- myoblasts was significantly promoted along with activated ERK1/2 and Cyclin D expression; the myogenic differentiation was impaired accompanied by the enhanced caspase 12/3 activation, implying enhanced endoplasmic reticulum (ER) stress. Interestingly, the bradykinin-induced Ca2+ release from ER calcium store was significantly enhanced after TMEM16A deletion. This suggested a suppressing role of intracellular TMEM16A in ER calcium release whereby regulating the flux of chloride ion across the ER membrane. Our findings reveal a unique location pattern of TMEM16A in undifferentiated myoblasts and its role in myogenesis.
Insights
Transmembrane protein 16A (TMEM16A) has a novel intracellular role in myoblasts, impacting skeletal muscle development by regulating endoplasmic reticulum calcium release and chloride ion flux.
Area of Science:
- Cell Biology
- Molecular Biology
- Muscle Development
Background:
- Transmembrane protein 16A (TMEM16A) is known to localize at the plasma membrane and regulate chloride influx.
- Its precise role and localization within muscle precursor cells (myoblasts) remain less understood.
Purpose of the Study:
- To investigate the intracellular localization and function of TMEM16A in myoblasts.
- To elucidate the role of TMEM16A in skeletal muscle development and myogenesis.
Main Methods:
- Immunofluorescence microscopy to determine TMEM16A localization in myoblasts.
- Genetic deletion of TMEM16A in mice and subsequent in vitro analysis of myoblast proliferation and differentiation.
- Assessment of signaling pathways (ERK1/2, Cyclin D), apoptosis markers (caspase 12/3), and endoplasmic reticulum (ER) calcium release.
Main Results:
- TMEM16A was found to localize primarily in the cytosolic compartment of myoblasts, partially co-localizing with intracellular organelles.
- Global deletion of TMEM16A resulted in severe skeletal muscle developmental defects.
- Tmem16a-/- myoblasts exhibited increased proliferation, impaired myogenic differentiation, enhanced ER stress, and elevated bradykinin-induced Ca2+ release from ER stores.
- Intracellular TMEM16A appears to suppress ER calcium release by regulating chloride ion flux across the ER membrane.
Conclusions:
- TMEM16A exhibits a unique intracellular localization pattern in undifferentiated myoblasts.
- Intracellular TMEM16A plays a crucial role in regulating ER calcium homeostasis and chloride ion transport, thereby influencing myogenesis and skeletal muscle development.
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