Andersen’s syndrome mutants produce a knockdown of inwardly rectifying K+ channel in mouse skeletal muscle in vivo

Abstract

Andersen's syndrome (AS) is a rare autosomal disorder that has been defined by the triad of periodic paralysis, cardiac arrhythmia, and developmental anomalies. AS has been directly linked to over 40 different autosomal dominant negative loss-of-function mutations in the KCNJ2 gene, encoding for the tetrameric strong inward rectifying K channel K2.1. While K2.1 channels have been suggested to contribute to setting the resting membrane potential (RMP) and to control the duration of the action potential (AP) in skeletal and cardiac muscle, the mechanism by which AS mutations produce such complex pathophysiological symptoms is poorly understood. Thus, we use an adenoviral transduction strategy to study in vivo subcellular distribution of wild-type (WT) and AS-associated mutant K2.1 channels in mouse skeletal muscle. We determined that WT and D71V AS mutant K2.1 channels are localized to the sarcolemma and the transverse tubules (T-tubules) of skeletal muscle fibers, while the ∆314-315 AS K2.1 mutation prevents proper trafficking of the homo- or hetero-meric channel complexes. Whole-cell voltage-clamp recordings in individual skeletal muscle fibers confirmed the reduction of inwardly rectifying K current (I) after transduction with ∆314-315 K2.1 as compared to WT channels. Analysis of skeletal muscle function revealed reduced force generation during isometric contraction as well as reduced resistance to muscle fatigue in extensor digitorum longus muscles transduced with AS mutant K2.1. Together, these results suggest that K2.1 channels may be involved in the excitation-contraction coupling process required for proper skeletal muscle function. Our findings provide clues to mechanisms associated with periodic paralysis in AS.

Ion Channel Contributions to Morphological Development: Insights From the Role of Kir2.1 in Bone Development

“…Mutations that disrupt the inwardly rectifying potassium channel (Kir2.1), encoded by the KCNJ2 gene, cause Andersen-Tawil syndrome. Patients with this syndrome have higher incidence of digital abnormalities, as well as cleft palate and other craniofacial abnormalities (Nguyen et al, 2013;Simkin et al, 2018). Cell culture work using Andersen Syndrome induced pluripotent stem cell mesenchymal stem cells (AS-IPSC-MSCs) revealed lower chondrogenic differentiation potential compared to wild type cells (Sacco et al, 2015;Pini et al, 2018).…”