2002 Fiscal Year Final Research Report Summary
Adencviral vector-mediatedin vivogene transfer of neurotropnins for experimental spinal cord injury in rats
| Project/Area Number |
13470299
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Orthopaedic surgery
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| Research Institution | Chiba University |
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Principal Investigator |
YOSHINAGA Katsunori Chiba University, University Hospital, Associate Professor, 医学部附属病院, 助教授 (30270870)
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| Co-Investigator(Kenkyū-buntansha) |
MURATA Atsushi Chiba University, University Hospital, Assistant, 医学部附属病院, 助手 (20344997)
SHIRASAWA Hiroshi Chiba University, Graduate School of Medicine, Professor, 大学院・医学研究院, 教授 (00216194)
YAMAZAKI Masashi Chiba University, University Hospital, Assistant, 医学部附属病院, 助手 (50281712)
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| Project Period (FY) |
2001 – 2002
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| Keywords | Spinal cord injury / brain-derived neurotrophic factor / Adenoviral vector |
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| Research Abstract |
The aim of this study is to evaluate the effect of adenovirus vector-mediated gene transfer of brain-derived' neurotrophic factor (BDNF) on axonal regeneration after spinal cord injury. We prepared adenovirus vectors encoding either β-galactosidase (AxCALacZ) or BDNF (AxCABDNF). BDNF produced by the adenovirus was detected by Western blotting and the bioactivity of recombinant BDNF was confilmed by bioassay. AxCALacZ was used to assess infection levels of the adenovirus. As a model of spinal cord injury, the rat spinal cord was completely transected at the T8 level. Immediately after the transection, the vectors were injected into both stumps. Axonal regeneration after transaction was assessed by retrograde and anterograde tracing. In AxCALacZ-injected rats, adenovirus-infected cells were observed not only in the injected site but also in the brainstem nuclei, shown by LacZ expression. After the injection of the retrograde tracer fluoregold (FG) in the distal portion to the transacted site, AxCABDNF-injected rats showed FG-labeled neurons in the red nucleus. The anterograde tracer biotinylated dextran amine (BDA) injected into the red nucleus was also found in regenerating rubrospinal fibers at the distal portion to the transected site. These tracing experiments revealed the regeneration of descending axons. In addition, Rats of the AxCABDNF group showed significant locomotor recovery of hindlimb function, which was completely abolished by re-transection. These results indicate that this recovery was caused by the rubrospinal axonal regeneration, not by the simple enhancement of central pattern generator.
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