The present study was performed to investigate the alternation of galanin in nociceptive modulation and galanin-like immunoreactivity in the central nervous system of rats after morphine tolerance. The hindpaw withdrawal latencies to both thermal and mechanical stimulation increased significantly after intracerebroventricular injection of 3 nmol of galanin in opioid-naive rats. The antinociceptive effect induced by galanin was attenuated remarkably at the same dose in morphine-tolerant rats. Furthermore, an up-regulation of galanin-like immunoreactivity in the arcuate nucleus of hypothalamus of morphine-tolerant rat was observed by immunohistochemical methods, whereas no significant changes were detected in periaqueductal gray. The present study demonstrated that there are alternations in both galanin-induced antinociception and galanin-like immunoreactivity in the brain of rat after morphine tolerance. The results suggest an involvement of galanin in the central nervous system in morphine tolerance.