| At present, there is no universally accepted treatment for paraplegia caused by injury to spinal cord, mianly because of the innocent of many genes and their functions believed to be involved in the complex pathophysiology of SCI. From the batch of upregulated EST sequences isolated by subtractive hybridization from full-transected spinal cord, this job tries to dig out more new genes and hints of their functions underlying injury and regeneration of spinal cord. Forty genes, among which twenty are new, were recognized from these ESTs. Functional annotation showed that they were most possibly engaged in widespread cell processes, which displayed again the complexity of spinal cord injury and regeneration. Among all these genes, Scirrl encoded a protein bearing both F-box and LRRs motifs, which is the typical motif structure of F-box proteins that recruit substrates for ubiquitination by E3 ubiquitin ligase complexes. Study of Scirrl may lead to the discovery of new molecules regulated by ubiquitin/proteasome pathway. Aided by bioinformatics, the coding region of Scirrl has been successfully cloned and expressed. Then the Dig-labeled riboprobe and rabbit antiserum were also prepared for further use. The up-regulation of SCIRR1 protein in spinal cord after full-transection was confirmed by Western Blot and immunohistochemistry. The whole-mount in situ hybridization of embryos showed the involvement of Scirrl in development of CNS. By in situ hybridization, Scirrl was also detected mainly in CNS and intestine, while little in pancreas, spleen and lung. Detailed analysis by in situ hybridization and immunohistochemistry showed a strong upregulation of Scirrl mRNA and protein in brain somatomotor area, hippocampus and gray matter of spinal cord especially the ninth layer neurons. The molecular cloning and characterization of Scirrl makes a good start on its functional research.
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