Insulin resistance (IR) in skeletal muscle is a pre-requisite for type 2 diabetes and is often associated with obesity. IR also develops alongside muscle atrophy in older individuals in sarcopaenic obesity. The molecular defects that underpin this syndrome are not well characterised and there is no licenced treatment. Deletion of the transforming growth factor-β family member myostatin, or sequestration of the active peptide by overexpression of the myostatin propeptide/ latency associated peptide (ProMyo), results in both muscle hypertrophy and reduced obesity and IR. We aimed to establish whether local myostatin inhibition would have a paracrine/autocrine effect to enhance glucose disposal beyond that simply generated by increased muscle mass, and the mechanisms involved. We directly injected adeno-associated virus expressing ProMyo into right tibialis cranialis/ extensor digitorum longus muscles of rats and saline into left muscles and compared the effects after 17 days. Both test muscles were increased in size (by 7% and 11%) and showed increased radiolabelled 2-deoxyglucose uptake (26% and 47%) and glycogen storage (28% and 41%) per unit mass during an intraperitoneal glucose tolerance test. This was likely mediated through increased membrane protein levels of GLUT1 (19% higher) and GLUT4 (63% higher). Interestingly, phosphorylation of phosphoinositol 3-kinase signalling intermediates and AMP-activated kinase were slightly decreased, possibly due to reduced expression of insulin-like growth factor-1 in these muscles. Thus, myostatin inhibition has direct effects to enhance glucose disposal in muscle beyond that expected of hypertrophy alone and this approach may offer potential for the therapy of IR syndromes.