|Budget Amount *help
¥3,900,000 (Direct Cost: ¥3,900,000)
Fiscal Year 2002: ¥1,900,000 (Direct Cost: ¥1,900,000)
Fiscal Year 2001: ¥2,000,000 (Direct Cost: ¥2,000,000)
We used a couple of neural survival factor, BDNF (brain-derived neurotrophic factor) and CNTF (cilialy neurotrophic factor) provided by investigation of the culture system of adult mammalian retina in last year, and confirm their effects by in vivo analysis. Retinal ganglion cells of adult rat were pre-labeled by retrograde labeling of fluorescent dye (Dil) and used for examination. The left optic verve was completely cut intra-orbitally under an anesthesia, and both optic nerve stump were bridged by a silicon tube which packed with collagen gel. Two weeks after the operation, the retina was isolated after an extraction of eyeball, and the labeled retinal ganglion cells were observed and measured the number of the living cells under a fluorescent microscope. Furthermore, after perfusion fixation, the optic nerve-optic nerve joint and a regenerated axons were examined about various kinds of materials (c-jun, c-fos, GAP-43, L1, etc.) related to neuronal cell death and regeneration by immunohistochemistry. In order to confirm whether a regenerative axon grow to superior colliculus across a gap of the optic nerve in a silicon tube, other fluorescent dye (fast blue) was injected to distal optic nerve. Then the number of double-labeld cells were counted in the flat-mounted retina. As a result what both factor acted on as a preventive factor and a neurotrophic factor on neural cell death of an adult retina in vivo same as a result provided from the retinal culture system. In addition, a novel factor, liver-derived neuronal activator, and the other neurotrophic factors which we previously found out and promote axonal regeneration similarly, and are to investigate possibility to be useful in functional reconstruction of a visual system.