Physical inactivity alters glucose homeostasis in skeletal muscle, potentially developing into overt metabolic disease. The present study sought to investigate the role of skeletal muscle capillarization in glucose tolerance and insulin sensitivity (IS) using a classic human model of physical inactivity. Thirteen healthy males (age=23±2 yr) underwent 4 days of full-time supervised and diet-controlled bed rest. Oral glucose tolerance test (OGTT), indices of insulin sensitivity (IS) (quantitative IS check index (QUICKI), Matsuda index) as well as skeletal muscle biopsies with measurement of fiber type distribution, fiber cross-sectional area (FCSA), capillary-to-fiber ratio (C/F ratio) and capillary density (CD) were assessed prior to and after bed rest. Body weight and composition were unaltered by bed rest. Fasting glucose/insulin ratio (G₀/I₀ ratio) (-25 %, P=0.016), QUICKI (-7 %, P=0.023) and Matsuda index (-24 %, P=0.003) diminished with bed rest. Skeletal muscle FCSA decreased (-737.4±763.2 µm^-2 (-12 %), P=0.005) while C/F ratio was preserved, resulting in augmented CD (+93.9±91.5 capillaries•mm^-2 (+37 %), P=0.003) with bed rest. No association was detected between changes in skeletal muscle variables and metabolic outcomes. Independently of bed rest-induced effects, a positive linear relationship was detected between C/F ratio and G₀/I₀ ratio (β=17.09, P=0.021). In conclusion, impaired glucose homeostasis with bed rest is not prevented nor associated with enhanced skeletal muscle capillarization in healthy individuals.