Objective To clone injury-regeneration-related genes after primary neurons of spinal cord were injured in rat, and to elucidate the molecular mechanism of injury and regeneration in CNS. Methods An in vitro spinal cord primary neuron injury model was replicated in rat. The differentially expressed genes during injury and regeneration periods of spinal cord primary neurons were cloned by improved subtractive hybridization. Results Twenty-seven differentially expressed sequences were obtained, 23 were known genes and the others were unknown. Among these known sequences, cDNA for synuclein and clusterin, together with several other gene sequences, were closely related with nerve degenerative diseases. Conclusion Primary neuron injury-regeneration-related genes of the spinal cord can be successfully cloned by improved subtractive hybridization.

Spinal cord injury and regeneration involves transcriptional activity of many genes, of which many remain unknown. Using the rat spinal cord full- transection model, bioinformatics, cloning, expression assays, fusion proteins, and transfection techniques, we identified and characterized one such differentially expressed gene, termed scirr1 (spinal cord injury and/or regeneration related gene 1). Fourteen orthologs were found in 13 species from echinoderm to insect and human by Blast search of NCBI protein reference sequence database. However, no further information is available for these homologues. Using whole-mount in situ hybridization, mouse scirr1 mRNA was expressed temporally and spatially in accordance with the early development sequence of the central nervous system. In adult rat spinal cord, expression of scirr1 mRNA was localized to neurons of gray matter by in situ hybridization. Using immunohistochemistry, SCIRR1 protein was found to be up-regulated and expressed more highly in spinal cord neurons farther from the epicenter of injury. Although the precise function of SCIRR1 is unknown, its unique pattern of expression during CNS early development and up-regulation after spinal cord injury suggest that SCIRR1 should be involved in the succeeding injury and/or repair processes of the injured spinal cord. Also, the typical F-box and leucine-rich repeat (LRR) architecture of rat SCIRR1 indicated that it may play an important substrate recruiting role in the pleiotropic ubiquitin/proteasome pathway. All these make scirr1 a new interesting start to study the spinal cord injury and regeneration mechanism.
Amino Acid Sequence ; Animals ; Base Sequence ; Brain/embryology/metabolism ; Embryo, Mammalian/metabolism ; F-Box Proteins/*biosynthesis ; Gene Expression Regulation, Developmental ; Male ; Mice ; Molecular Sequence Data ; Organ Specificity ; PC12 Cells ; Phylogeny ; Rats ; Rats, Wistar ; Spinal Cord/embryology/metabolism ; Spinal Cord Injuries/*metabolism ; Up-Regulation