In isolated resistance arterioles with spontaneous tone, ligation of α₄β₁- and α₅β₁-integrins induces vasoconstriction whereas ligation of α_vβ₃-integrin induces vasodilation. However, whether integrins directly participate in myogenic constriction to pressure elevation is not known. To answer this question, isolated rat skeletal muscle arterioles were exposed to step increments in pressure in the absence or presence of peptides and function-blocking antibodies known to bind α₄β₁-, α₅β₁-, or α_vβ₃-integrins while vessel diameter was continually monitored. Myogenic constriction, as assessed by the ability of isolated arterioles to reduce their diameter in response to two consecutive increments in intraluminal pressure (90–110 and 110–130 cmH₂O), was not affected by treatment with any of the control peptides (RAD, LEV), a control antibody (anti-rat major histocompatibility complex), an α₄β₁-integrin-binding peptide (LDV), or an anti-α₄-integrin antibody. In contrast, α₅β₁-integrin blockade with either anti-α₅- or anti-β₁-integrin antibody caused a significant inhibition of myogenic constriction. Also, both RGD peptide and anti-β₃-integrin antibody inhibited myogenic constriction. These results indicate that α₅β₁- and α_vβ₃-integrins are necessary for myogenic constriction and further suggest that integrins are part of the mechanosensory apparatus responsible for the ability of vascular smooth muscle cells to detect and/or respond to changes in intraluminal pressure.