The present study was designed to investigate the role of glycine receptors in analgesia induced by injection of glycine in vivo. Glycine was injected intracerebroventricularly or intrathecally and strychnine, a glycine receptor antagonist, was injected intracerebroventricularly or intrathecally before glycine injection. The effects on the pain threshold index in hot-plate test and the writhing times in acetic acid-induced writhing test were observed. The locomotor activity and motor performance (rotarod test) were also observed. The dosages of glycine and strychnine we choose had no effect on locomotor activity or motor performance in conscious mice. Glycine increased the pain threshold index in hot-plate test and decreased the writhing times of the mice. Strychnine antagonized the effects induced by glycine above. These results demonstrated that intracerebroventricular or intrathecal injection of glycine can produce analgesia in thermal nociception and chemical nociception in vivo, which is mediated by glycine receptors.