|Budget Amount *help
¥2,100,000 (Direct Cost : ¥2,100,000)
Fiscal Year 1994 : ¥400,000 (Direct Cost : ¥400,000)
Fiscal Year 1993 : ¥600,000 (Direct Cost : ¥600,000)
Fiscal Year 1992 : ¥1,100,000 (Direct Cost : ¥1,100,000)
There is a discrepancy between the data in vitro and that in vivo concerning nerve promoting factors. This is because that in vivo system might contain some additional factors which were not suspected in vitro. Then we intend to establish a new experimental system in vivo using killed nerve graft and/or DRG-transplantation, which will make possible to analyze the mechanism of peripheral nerve regeneration under in vivo condition.
Because of the difficulty of obtaining neurotrophic reagents, BDNF and CNTF,only a few experiments using these neurotrophic factors were performed. However, our experiments revealed that immunostained teased specimens are useful for the observation under confocal laser scanning microscope. Additional important findings are as follows ;
(1) Several microns to the tips of regenerating axons growing through basal lamina tubes in vivo exhibited immunoreactivities for GAP43, PGP9.5 and synaptophysin, indicating that these region might have some growth cone-like functions as in development or in culture.
(2) Fifth day after the injection of various tracers, i.e., Dil, WGA-HRP,Fast Blue, and fluorescent latex microspheres, into rat sciatic nerve, dense accumulation of Dil in satellite cells associated with large and small Dil-labeled DRG cells was visible. It indicates a possibility of molecular transport from DRG neurons to satellite cells in mammalian nervous system.
(3) In normal rat spinal cord and facial nucleus, residential ramified microglial cells were immunoreactive for thymosin and OX-42 in both gray and white matter. The morphological characteristics of thymosin-immunoreactive cells correspond with those of Hortega cells, indicating that this anti-thymosin antibody is a good marker for microglia.