The aim of this study was to measure vascular reactivity in the isolated middle cerebral artery (MCA) after brain injury. Segments of MCA were prepared from control, sham-operated, and cold-lesioned rats. Cold lesion was induced by application of a precooled (-78 degrees C) copper cylinder (diameter 5 mm) for 60 sec to the intact dura over the parietal cortex. Endothelin-1 (ET-1) (10(-12) to 3 x 10(-7) M) induced a dose-dependent contraction with a pD2 (-log10 EC50) of 8.36+/-0.12 (mean+/-SEM) and an Emax (maximal response) of 2.41+/-0.15 mN (millinewton) at 10(-7) M in sham-operated animals under resting conditions. This maximum contraction induced by 10(-7) M ET-1 was significantly (p < 0.05) reduced 24 and 48 h after cold lesion by 41% and 30%, respectively. After precontraction with 10(-5) M prostaglandin (PG) F2alpha, ET-3 (10(-12) to 10(-8) M) relaxed the MCA with an Emax of 0.42+/-0.07 mN at 10(-8) M and a pD2 of 9.20+/-0.19 in sham-operated animals. This relaxation was reduced 24 and 48 h after cold lesion by 19% and 62% at 10(-8) M, respectively. Concentration-effect curves for bradykinin (BK, 10(-8) to 10(-5) M) in uridine triphosphate (UTP, 10(-4) M)-precontracted MCA segments revealed relaxation with a pD2 of 7.08+/-0.10 and an Emax of 0.65+/-0.06 mN at 10(-6) M in sham-treated animals. This effect of BK was reduced by 35% and 20% at 10(-6) M 24 and 48 h after cold lesion, respectively. In addition, the contractile responses to 124 mM K+, 10(-5) M PGF2alpha and the dilation induced by 10(-3) sodium nitroprusside (SNP) were reduced in MCA segments taken 24 and 48 h after lesion compared with shams. We conclude that attenuation of ET effects can be explained, at least in part, by tachyphylaxis to ETs. The unspecific reduction of vascular reactivity may result from spreading depression.