c-fos expression in isolated rat spinal cord

Expression of c-fos-immunoreactivity (c-fos-ir) has been demonstrated in the dorsal horn of lumbar segments of an isolated spinal cord preparation from 3 week old rats. The method of preparation generated a low level of c-fos-ir activity which was not significantly altered by low intensity (1.5 times threshold) dorsal root stimulation, but was significantly increased by high intensity (20 times threshold) stimulation. Replacement of the calcium in the bathing medium by 2 mM manganese suppressed all detectable c-fos-ir, whereas inclusion of 0.5 microM capsaicin caused intense c-fos-ir expression in the absence of stimulation. The number of dorsal horn cells exhibiting c-fos-ir increased between 0.5 and 1 h after stimulation, reaching a maximum at 2 h, with no further increase at longer intervals. Few positive cells were found when the incubation temperature was reduced from 27 to 20 degrees C. The strongest increase in c-fos-ir was found in the dorsal horn ipsilateral to the stimulated dorsal root and a smaller, but significant, increase was also seen in the contralateral dorsal horn. Cords obtained from animals treated at 1 day old with capsaicin to destroy afferent C fibres showed a reduction in the number of c-fos-ir positive cells induced by high intensity dorsal root stimulation. This preparation will aid detailed investigation of the pharmacology of nociceptive pathways.