| Project/Area Number |
10470005
|
|---|
| Research Category |
Grant-in-Aid for Scientific Research (B)
|
| Allocation Type | Single-year Grants |
|---|
| Section | 一般 |
|---|
| Research Field |
General anatomy (including Histology/Embryology)
|
|---|
| Research Institution | Kyoto University |
|---|
Principal Investigator |
IDE Chizuka Kyoto Univ.Grad.School of Medicine, Professor, 医学研究科, 教授 (70010080)
|
|---|
| Co-Investigator(Kenkyū-buntansha) |
FUJIMOTO Kazushi Kyoto Univ.Grad.School of Medicine, Lecturer, 医学研究科, 講師 (50159125)
MIZOGUCHI Akira Kyoto Univ.Grad.School of Med, Associate Professor, 医学研究科, 助教授 (90181916)
KAWAGUCHI Saburo Kyoto Univ.Grad.School of Medicine, Professor, 医学研究科, 教授 (70024635)
NODA Tohru Kyoto Univ.Grad.School of Medicine, Instructor, 医学研究科, 助手 (50156204)
早柏 琢哉 京都大学, 医学研究科, 助手 (00273459)
|
|---|
| Project Period (FY) |
1998 – 1999
|
| Project Status |
Completed (Fiscal Year 1999)
|
| Budget Amount *help |
¥12,500,000 (Direct Cost: ¥12,500,000)
Fiscal Year 1999: ¥2,900,000 (Direct Cost: ¥2,900,000)
Fiscal Year 1998: ¥9,600,000 (Direct Cost: ¥9,600,000)
|
|---|
| Keywords | nerve regeneration / choroid plexus / ependymal cell / graft / spinal cord / coculture / dorsal root ganglion neuron / axonal outgrowth |
|---|
| Research Abstract |
1 Grafting into spinal cord
The choroid plexus from the 4th ventricle of adult rats (Wistar) were minced into small fragments and grafted into the dorsal funiculus at the C2 level in adult rat spinal cord from the same strain. Electron microscopy and fluorescence histochemistry showed that ependymal cells of the grafted choroid plexus intimately interacted with growing axons, serving to support the massive growth of regenerating axons. HRP injection at the sciatic nerve showed that numerous HRP-labeled regenerating fibers from the fasciculus gracilis extended into the graft 7 days after grafting. These regenerating axons from the fasciculus gracilis were maintained for at least 10 months, with some axons elongating rostrally into the dorsal funiculus. Evoked potentials of long duration were recorded at a level ca. 5mm rostral to the lesion in the rats 8 to 10 months after grafting. These findings indicate that choroid plexus ependymal cells have the ability to facilitate axonal growth in vivo, suggesting that they may be a promising candidate as graft for the promotion of nerve regeneration in the spinal cord.
2 Coculture experiment
Choroid plexuses from the 4th ventricle of postnatal day-1 to -5 mice, were mechanically dissociated and plated in bronectin-coated culture dishes. Ependymal cells spread in monolayer with few endothelial cells at 3-week culture. Dorsal root ganglia (DRG) were cocultured on the choroid plexus ependymal cell monolayer. It was demonstrated that neurons extended many long neurites with elaborate branchings on the surface of S100 β-stained ependymal cells. In contrast, DRG neurons cultured on the astrocytes and laminin-coated plates had much fewer and shorter primary neurites with fewer branches than those cultured on the ependymal cells. These results suggest that choroid plexus ependymal cells have a great capacity to promote neurite outgrowth from DRG neurons in vitro.
|
