To assess the effects of microvascular temporary clip application on vessel relaxing capability and endothelial substance release, the carotid rings of rats clipped for various durations were studied via bioassay. Noradrenaline and phenylephrine produced an immediate contraction and subsequent relaxation that failed to be suppressed by lysine acetylsalicylate or nicotine in the controls and in the arteries clipped for 0.5, 1, and 5 minutes; however, this relaxation was greatly reduced when the duration was 10 minutes. The results suggest the possible role of inadequate endothelium-derived relaxing factor release following prolonged application of the clips.