Modulation of gene expression during early stages of reconnection of the turtle spinal cord

The spinal cord of the freshwater turtle Trachemys dorbignyi regenerates after complete transection (Rehermann et al. J. Comp. Neurol. 515, 2009, 197-214). This remarkable ability may be related to the persistence around the central canal (CC) of progenitors functionally clustered via connexin 43 (Cx43) that express brain lipid binding protein (BLBP) and the transcription factor Pax6 (Russo et al. J. Neurosci. 28, 2008, 8510-8516). Indeed, because BLBP+ cells appear in the bridge joining the rostral and caudal stumps, we speculated that progenitors contacting the central canal may play a key part in spinal cord regeneration. To test this hypothesis, we designed degenerated primers pairing conserved regions for key proteins synthesized in progenitors (BLBP, Cx43, and Pax6) and the neuronal protein HuB. Fragments of these proteins were amplified, cloned, and sequenced. Based on these sequences, we analyzed the changes in the expression levels using quantitative real-time RT-PCR with specific primers, comparing the injured spinal cord at different times after injury (4, 12, 20, and 60 days) with uninjured spinal cords. We found a transient, early increase of BLBP, Cx43 and HuB mRNA, with Pax6 remaining unchanged. These results suggest that the selected genes--active in progenitor cells--play an important part in early mechanisms of spinal cord regeneration.