Related Experiment Video
Updated: Jul 2, 2025
Engineering Skeletal Muscle Tissues from Murine Myoblast Progenitor Cells and Application of Electrical Stimulation
Published on: March 19, 2013
A Pharmacological Investigation of the TMEM16A Currents in Murine Skeletal Myogenic Precursor Cells
Marina Sciancalepore1, Asja Ragnini1, Paola Zacchi1
1Department of Life Sciences, University of Trieste, I-34127 Trieste, Italy.
Abstract:
TMEM16A is a Ca2+-activated Cl- channel expressed in various species and tissues. In mammalian skeletal muscle precursors, the activity of these channels is still poorly investigated. Here, we characterized TMEM16A channels and investigated if the pharmacological activation of Piezo1 channels could modulate the TMEM16A currents in mouse myogenic precursors. Whole-cell patch-clamp recordings combined with the pharmacological agents Ani9, T16inh-A01 and Yoda1 were used to characterize TMEM16A-mediated currents and the possible modulatory effect of Piezo1 activity on TMEM16A channels. Western blot analysis was also carried out to confirm the expression of TMEM16A and Piezo1 channel proteins. We found that TMEM16A channels were functionally expressed in fusion-competent mouse myogenic precursors. The pharmacological blockage of TMEM16A inhibited myocyte fusion into myotubes. Moreover, the specific Piezo1 agonist Yoda1 positively regulated TMEM16A currents. The findings demonstrate, for the first time, a sarcolemmal TMEM16A channel activity and its involvement at the early stage of mammalian skeletal muscle differentiation. In addition, the results suggest a possible role of mechanosensitive Piezo1 channels in the modulation of TMEM16A currents.
Insights
Calcium-activated chloride channel TMEM16A is active in mouse skeletal muscle precursors and essential for myocyte fusion. Piezo1 channel activation enhances TMEM16A currents, suggesting a role in muscle development.
Area of Science:
- Ion channel physiology
- Skeletal muscle biology
- Cellular mechanotransduction
Background:
- TMEM16A (Anoctamin-1) is a calcium-activated chloride channel with diverse physiological roles.
- Its function in mammalian skeletal muscle precursor cells remains largely uncharacterized.
- Understanding these channels is crucial for insights into muscle development and regeneration.
Purpose of the Study:
- To characterize TMEM16A channel activity in mouse myogenic precursors.
- To investigate the modulatory effect of Piezo1 channel activation on TMEM16A currents.
- To determine the role of TMEM16A in skeletal muscle differentiation.
Main Methods:
- Whole-cell patch-clamp electrophysiology to record TMEM16A currents.
- Pharmacological agents (Ani9, T16inh-A01, Yoda1) to modulate TMEM16A and Piezo1 channels.
- Western blot analysis to confirm protein expression of TMEM16A and Piezo1.
Main Results:
- TMEM16A channels are functionally expressed in fusion-competent mouse myogenic precursors.
- Inhibition of TMEM16A channels significantly impaired myocyte fusion into myotubes.
- Activation of the mechanosensitive Piezo1 channel by Yoda1 enhanced TMEM16A currents.
Conclusions:
- This study provides the first evidence of sarcolemmal TMEM16A channel activity in early mammalian skeletal muscle differentiation.
- TMEM16A channels play a critical role in the fusion of myogenic precursors.
- Mechanosensitive Piezo1 channels may modulate TMEM16A channel activity during skeletal muscle development.
