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1 Acta Neurobiologiae Experimentalis

1 2011

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Intraspinal injection of AAV5_eGFP or AAV5_L1: extent of transduction and influence on regrowth of CST axons in adult rats after spinal cord transection

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Platek R. , Korczynski J. , Czarkowska-Bauch J. , Skup M.

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Among approaches targeting restoration of function after spinal cord injury a promising one is to use L1 cell adhesion molecule, known to promote axon outgrowth, fasciculation, guidance and myelination in regeneration. L1 is upregulated after injury, manifesting requirements of the impaired networks for successful repair. Previously, our strategy to overexpress L1 gene in the lesion site was found to support reinnervation of the lumbar spinal cord after thoracic spinal cord compression in mice. Here we verified whether L1 overexpression induced caudal to complete spinal transection, may still have an impact on rostral segments affecting a regrowth of corticospinal tract (CST). AAV5 vector encoding eGFP or L1 protein under mCMV promoter was injected bilaterally into spinal L1 segment 30min after spinal cord transection at Th10/11. To label CST, rats were injected with anterograde tracer DiI to sensorimotor cortex, 1 week after spinalization. Effectiveness of transduction with AAV5 vector was evaluated based on distribution of eGFP protein at 7, 14 and 35 days. To evaluate CST regrowth, distances between lesion rostral border and tips of regrowing/sprouting single CST axons as well as the majority of axons were measured. eGFP expression occured throughout entire dorsoventral axis, in spinal gray and white matter, already 7 days postlesion and maintained up to 35. eGFP (+) fibers were traversing all segments caudally to lesion, some closing near its border but none seen in the lesion. It indicated that L1 transgene may be long-term available within segments below transection. CST tracing in AAV5_L1 revealed that in 3 out of 5 rats majority of CST axons reached the lesion border, whereas in AAV5_eGFP group only 1 out of 6 rats showed similar contact. In conclusion, AAV5_L1 overexpressed in segments caudal to complete transection may affect cellular milieu in transection proximity which results in better CST regrowth. Support: S007/Polish-German/2007/01 grant.