Effect of insulin insufficiency on ultrastructure and function in skeletal muscle

Background Decreased insulin availability and high blood glucose levels, the hallmark features of poorly controlled diabetes, drive disease progression and are associated with decreased skeletal muscle mass. We have shown that mice with β-cell dysfunction and normal insulin sensitivity have decreased skeletal muscle mass. This project asks how insulin deficiency impacts on the structure and function of the remaining skeletal muscle in these animals. Methods Skeletal muscle function was determined by measuring exercise capacity and specific muscle strength prior to and after insulin supplementation for 28 days in 12-week-old mice with conditional β-cell deletion of the ATP binding cassette transporters ABCA1 and ABCG1 (β-DKO mice). Abca1 and Abcg1 floxed (fl/fl) mice were used as controls. Expression of genes encoding for skeletal muscle atrophy markers and signaling pathways were quantified by RNA-seq expression profiling. Skeletal muscle and mitochondrial morphology were assessed by transmission electron microscopy. Sarcomere patterns were assessed by label-free Second Harmonic Generation microscopy. Myofibrillar Ca2+ sensitivity and maximum isometric single muscle fibre force were assessed using MyoRobot biomechatronics technology. Results 59 RNA transcripts involved in muscle contraction and movement were significantly altered in β-DKO mice compared to fl/fl controls. Exercise capacity and muscle strength were significantly decreased in β-DKO mice compared to fl/fl controls (p=0.012), and a loss of structural integrity was also observed in skeletal muscle from the β-DKO mice. Supplementation of β-DKO mice with insulin restored muscle integrity, strength and expression of 21 of the dysregulated transcripts. Conclusions Insulin insufficiency due to β-cell dysfunction perturbs the structure and function of skeletal muscle. These adverse effects are rectified by insulin supplementation.