Viral vector-mediated expression of K+ channels regulates electrical excitability in skeletal muscle

被引:12
作者
Falk, T
Kilani, RK
Yool, AJ
Sherman, SJ
机构
[1] Univ Arizona, Coll Med, Dept Physiol, Tucson, AZ 85724 USA
[2] Univ Arizona, Coll Med, Dept Neurol, Tucson, AZ 85724 USA
[3] Univ Arizona, Coll Med, Dept Pharmacol, Tucson, AZ 85724 USA
[4] Univ Arizona, Program Neurosci, Tucson, AZ 85724 USA
关键词
hyperkalemic periodic paralysis; anemone toxin II; herpes simplex 1amplicon system; green fluorescent protein; Kv1.4; primary muscle culture;
D O I
10.1038/sj.gt.3301539
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Modification of K+ currents by exogenous gene expression may lead to therapeutic interventions in skeletal muscle diseases characterized by alterations in electrical excitability. In order to study the specific effects of increasing outward K+ currents, we expressed a modified voltage-dependent K+ channel in primary cultured rat skeletal muscle cells. The rat Kv1.4 channel was expressed as an N-terminal fusion protein containing a bioluminescent marker (green fluorescent protein). Transgene expression was carried out using the helper-dependent herpes simplex 1 amplicon system. Transduced myoballs, identified using fluorescein optics and stud-led electrophysiologically with single-cell patch clamp, exhibited a greater than two-fold increase in K+ conductance by 20-30 h after infection. This increase in K+ current led to a decrease in membrane resistance and a 10-fold increase in the current threshold for action potential generation. Electrical hyperexcitability induced by the Na+ channel toxin anemone toxin II (1 mum) was effectively counteracted by overexpression of Kv1.4 at 30-32 h after transduction. Thus, virally induced overexpression of a voltage-gated K+ channel in skeletal muscle has a powerful effect in reducing electrical excitability.
引用
收藏
页码:1372 / 1379
页数:8
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