Phosphorylation of extracellular signal-regulated kinases 1/2 is predominantly enhanced in the microglia of the rat spinal cord following dorsal root transection

Abstract

The present study was initiated to investigate the role of extracellular signal-regulated kinases (ERK) 1/2 signaling pathway in the early response of spinal cord and associated dorsal root ganglion (DRG) to rhizotomy by using Western blotting and immunohistochemical techniques in a rat model of L3 and L4 dorsal root transection. The results showed that there were a considerable amount of total and phosphorylated ERK 1/2 protein in both spinal cord and DRG in normal animals killed under pentobarbital anesthesia. The total ERK 1/2 distributed in both glia and neurons, while phosphorylated ERK 1/2 dominantly existed in the latter in the gray matter of spinal cord, as demonstrated with double immunofluorescent staining. Twenty-four and forty-eight hours after axotomy, the phosphorylation level of ERK 1/2 in the operation side of dorsal spinal cord was much higher than that in the contralateral side, while the total ERK 1/2 level seemed unchanged. The increased expression of Fos protein was also seen in the dorsal spinal cord at lesion side twelve and twenty-four hours after axotomy. Double fluorescent staining proved that the phosphorylated ERK 1/2 positive cells in the ipsilateral dorsal spinal cord after axotomy predominantly were microglia and small portion was oligodendrocytes, whereas the Fos expression was mainly in neurons. In normal DRG, most neurons, especially the medium and small-sized ones, and the satellite cells contained total ERK 1/2-like immunoreactivity, whereas only a small portion of neurons and satellite cells contained phosphorylated ERK 1/2. After unilateral dorsal rhizotomy, there were no detectable changes for the phosphorylation of ERK 1/2 in either neurons or satellite cells in DRG. Collectively, the present results suggest that both ERK and Fos signal pathways involve the cellular activation in the spinal cord following dorsal rhizotomy, with ERK mainly in microglia and Fos in neurons. The increase of phosphorylation of ERK 1/2 in microglia of spinal cord after rhizotomy implicates that ERK signaling pathway involves intracellular activity of microglia responding to the experimental injury.