Abstract

The activation of potassium channels and following hyperpolarization in skeletal myoblasts is essential for myogenic differentiation. However, the effects of K⁺ channel opening in myoblasts on skeletal muscle mass are unclear. Our previous study revealed that pharmacological activation of intermediate conductance Ca²⁺-activated K⁺ channels (IK_Ca channels) increases myotube formation. In this study, we investigated the effects of DCEBIO, a K_Ca channel opener, on the mass of skeletal muscle. Application of DCEBIO to C2C12 cells during myogenesis increased the diameter of C2C12 myotubes in a concentration-dependent manner. This DCEBIO-induced hypertrophy was abolished by gene-silencing of IK_Ca channels. However, it was resistant to 1 µM, but sensitive to 10 µM TRAM-34, a specific IK_Ca channel blocker. Furthermore, DCEBIO reduced the mitochondrial membrane potential by opening IK_Ca channels. Therefore, DCEBIO should increase myotube mass by opening of IK_Ca channels distributed in mitochondria. Pharmacological studies revealed that mitochondrial reactive oxygen species (mitoROS), Akt, and mTOR are involved in DCEBIO-induced myotube hypertrophy. An additional study demonstrated that DCEBIO-induced muscle hypertrophic effects are only observed when applied in the early stage of myogenic differentiation. In an in vitro myotube inflammatory atrophy experiment, DCEBIO attenuated the reduction of myotube diameter induced by endotoxin. Thus, we concluded that DCEBIO increases muscle mass by activating the IK_Ca channel/mitoROS/Akt/mTOR pathway. Our study suggests the potential of DCEBIO in the treatment of muscle wasting diseases.

Significance Statement Our study shows DCEBIO, a small molecule opener of Ca²⁺ activated K⁺ channel, increased muscle diameter via mitochondria ROS/Akt/mTOR pathway. And DCEBIO overwhelms C2C12 myotube atrophy induced by endotoxin challenge. Our report should inform novel role of K⁺ channel in muscle development and novel usage of K⁺ channel opener such as for the treatment of muscle wasting diseases.