Vascular endothelial growth factor (VEGF) is exercise-responsive, pro-angiogenic, and expressed in several muscle cell types. We hypothesized that in adult mice, VEGF generated within skeletal myofibers (and not other cells within muscle) is necessary for the angiogenic response to exercise training. This was tested in adult conditional, skeletal myofiber-specific VEGF gene-deleted mice (skmVEGF-/-), with VEGF levels reduced by >80%. After 8 weeks of daily treadmill training, speed and endurance were unaltered in skmVEGF-/- mice, but increased by 18% and 99% (p < 0.01), respectively, in controls trained at identical absolute speed, incline, and duration. In vitro, isolated soleus and EDL contractile function was not impaired in skmVEGF-/- mice. However, training-induced angiogenesis was inhibited in plantaris (WT, 38%, skmVEGF-/- 18%, p < 0.01), and gastrocnemius (WT, 43%, p < 0.01; skmVEGF-/-, 7%, n.s.). Capillarity was maintained (different from VEGF gene deletion targeted to multiple cell types) in untrained skmVEGF-/- mice. Arteriogenesis (smooth muscle actin+, artery number and diameter) and remodeling (vimentin+, BrDU+ and F4/80+ cells) occurred in skmVEGF-/- mice, even in the absence of training. SkmVEGF-/- mice also displayed a limited oxidative enzyme (citrate synthase and β -HAD) training response; β-HAD activity levels were elevated in the untrained state. These data suggest that myofiber expressed VEGF is necessary for training responses in capillarity and oxidative capacity, and for improved running speed and endurance.