Intense cold noxious stimulation of the rat hindpaw induces c-fos expression in lumbar spinal cord neurons.

This study evaluated Fos-like immunoreactivity in lumbar spinal cord neurons following intense cold stimulation and then the modifications induced by opioid administration. Under urethane anaesthesia, the rat's right foot was stimulated by holding it in a regulated temperature bath at 15, 10, 0, -10, -15, -17.5 or -20 degrees C. There was no or little Fos-like immunoreactivity in lumbar spinal cord neurons when the paw was at temperatures between 15 and -10 degrees C (0-5 Fos-like immunoreactive neurons/section). The threshold to induce consistent c-fos expression was -15 degrees C. From -15 to -20 degrees C, the number of Fos-like immunoreactive neurons increased with decreases in temperature. At -20 degrees C, Fos-like immunoreactive neurons were numerous in L3 and L4 segments, in laminae I-II (approximately 60 Fos-like immunoreactive neurons/section) and to a lesser extent in laminae V-VI (approximately 20). Almost no Fos-like immunoreactivity was present in laminae III-IV (< 5). At -20 degrees C, the number of Fos-like immunoreactive neurons increased with the duration of the stimulation. The number of Fos-like immunoreactive neurons induced by the cold stimulation temperatures was significantly decreased by pretreatment with 10 mg/kg s.c. morphine and moderately decreased by 5 mg/kg s.c. This effect was antagonized by the combined administration of morphine (10 mg/kg s.c.) and naloxone (2 mg/kg s.c.). Naloxone (2 mg/kg s.c.) significantly increased the number of Fos-like immunoreactive neurons induced by -20 degrees C as compared to saline-injected rats. This study showed that Fos-like immunoreactivity distribution is in good agreement with the location of neurons receiving noxious inputs and that the threshold to induce c-fos expression with cold was unexpectedly low at -15 degrees C. Taking into account, on the one hand, previous investigations using the same technique using noxious heat stimulation and, on the other hand, electrophysiological and psychophysiological studies using cold stimulation in animals and humans, our results suggest that Fos-like immunoreactivity induced by extremely cold stimulation, which seems to reproduce frostbite, may reflect activation of nociceptors due to vasoconstriction.