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Systemic Delivery of MicroRNA Using Recombinant Adeno-associated Virus Serotype 9 to Treat Neuromuscular Diseases in Rodents
Published on: August 10, 2018
MicroRNA is a potential target for therapies to improve the physiological function of skeletal muscle after trauma
Xin-Yi Gu1, Bo Jin2, Zhi-Dan Qi1
1Department of Orthopedics and Traumatology, Peking University People's Hospital; Key Laboratory of Trauma and Neural Regeneration (Peking University), Beijing, China.
Abstract:
MicroRNAs can regulate the function of ion channels in many organs. Based on our previous study we propose that miR-142a-39, which is highly expressed in denervated skeletal muscle, might affect cell excitability through similar mechanisms. In this study, we overexpressed or knocked down miR-142a-3p in C2C12 cells using a lentivirus method. After 7 days of differentiation culture, whole-cell currents were recorded. The results showed that overexpression of miR-142a-3p reduced the cell membrane capacitance, increased potassium current density and decreased calcium current density. Knockdown of miR-142a-3p reduced sodium ion channel current density. The results showed that change in miR-142a-3p expression affected the ion channel currents in C2C12 cells, suggesting its possible roles in muscle cell electrophysiology. This study was approved by the Animal Ethics Committee of Peking University in July 2020 (approval No. LA2017128).
Insights
MicroRNA-142a-3p (miR-142a-3p) influences muscle cell excitability by altering ion channel function. Its expression changes affect potassium, calcium, and sodium currents in C2C12 cells.
Area of Science:
- Molecular Biology
- Cell Physiology
- Muscle Electrophysiology
Background:
- MicroRNAs (miRNAs) are key regulators of gene expression, impacting cellular functions.
- Previous research suggests miRNAs affect ion channel activity.
- miR-142a-3p is highly expressed in denervated skeletal muscle, hinting at a role in muscle excitability.
Purpose of the Study:
- To investigate the role of miR-142a-3p in regulating ion channel function and electrophysiology in skeletal muscle cells.
- To determine how altering miR-142a-3p expression affects ion channel currents in C2C12 myotubes.
Main Methods:
- C2C12 mouse myoblast cells were cultured and differentiated for 7 days.
- miR-142a-3p was overexpressed or knocked down using lentiviral vectors.
- Whole-cell patch-clamp electrophysiology was employed to record ion channel currents.
Main Results:
- Overexpression of miR-142a-3p led to decreased cell membrane capacitance.
- Increased potassium current density and decreased calcium current density were observed upon miR-142a-3p overexpression.
- Knockdown of miR-142a-3p resulted in reduced sodium ion channel current density.
Conclusions:
- Modulating miR-142a-3p expression significantly impacts ion channel currents in C2C12 cells.
- These findings suggest miR-142a-3p plays a crucial role in skeletal muscle cell electrophysiology.
- miR-142a-3p represents a potential target for understanding and treating muscle excitability disorders.
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