Electroporated GLUT4-7myc-GFP detects in vivo glucose transporter 4 translocation in skeletal muscle without discernible changes in GFP patterns.

摘要: New findings What is the central question of this study? Resolving mechanism(s) leading to glucose transporter 4 (GLUT4) translocation muscle surface membrane has great therapeutic potential. However, measurement GLUT4 technically challenging. Here, we asked whether electroporation GLUT4-7myc-GFP into skeletal could be used as a tool study in vivo. main finding and its importance? By acutely inducing expression muscle, verified that vivo exercise AICAR stimulation increased presence sarcolemma measured myc signal. Importantly, signal was not accompanied by major visual changes distribution GFP Abstract Insulin lead fibres. This process pivotal for facilitating uptake muscle. To this, robust assay needed measure adult directly. aimed validate simple using genetically encoded biosensor mouse We transfected live movement evaluate insertion after running pharmacological activation AMP-activated protein kinase (AMPK). Transfection led dynamic flexor digitorum brevis fibres which, upon insulin stimulation, exposed epitope extracellularly. Running exercise, addition AMPK 5-aminoimidazole-4-carboxamide ribonucleotide, induced ∼125 ∼100% increase, respectively, extracellularly exposure tibialis anterior Interestingly, clear increase surface-exposed content insulin, or any discernible reorganization GLUT4-GFP In conclusion, provide detailed description an easy-to-use accumulation at exercise-mimicking stimuli. Notably, our analyses revealed change localization pattern.