To characterize the vasodilatory response in the transition from a single skeletal contraction to a series of contractions, we measured the response of hamster cremaster muscle arterioles associated with 4-5 skeletal muscle fibres stimulated to contract for 1, 2, 3, or 4 contractions (250ms train duration) at 4s intervals (15 contractions per minute (CPM)) for up to 12s, at stimulus frequencies of 4, 10, 20, 30, 40, 60 and 80Hz. To investigate the contribution of contraction frequency we stimulated muscle fibre bundles at 30 or 60CPM for 12s at stimulus frequencies of 4, 20 and 60Hz. Arteriolar diameters at the site of overlap with the stimulated muscle fibres were measured before and after each contraction. At 15CPM at 4, 20, and 60Hz, we observed a peak change in diameter following the first contraction of (1.1+/-0.1um, 1.6+/-0.2um, and 2.1+/-0.2um) that almost doubled in response to the second contraction (2.0+/-0.1um, 3.0+/-0.1um, and 3.8+/-0.1um respectively) but there was no further dilation following the third or fourth contraction. A similar response occurred at all stimulus and contraction frequencies tested. At 30 and 60CPM at 60Hz the plateau after 2 contractions was followed by a further increase in diameter to a second plateau at 7-8 seconds. Therefore, the vasodilatory response in the transition from single to multiple contractions had components that were stimulation parameter-dependent and -independent and showed a plateauing behavior indicative of rapid changes in either the nature and/or concentration of vasodilators released or changes in vascular reactivity.