| Project/Area Number |
01480126
|
|---|
| Research Category |
Grant-in-Aid for General Scientific Research (B)
|
| Allocation Type | Single-year Grants |
|---|
| Research Field |
Neurophysiology and muscle physiology
|
|---|
| Research Institution | Osaka University |
|---|
Principal Investigator |
MURAKAMI Fujio Osaka University, Fac. of Engineering Sci., Professor, 基礎工学部, 教授 (20089882)
|
|---|
| Co-Investigator(Kenkyū-buntansha) |
KATSUMARU Hironobu Osaka University, Fac. of Engineering Sci., Assistant, 基礎工学部, 助手 (40183264)
小田 洋一 大阪大学, 基礎工学部, 講師 (00144444)
|
|---|
| Project Period (FY) |
1989 – 1990
|
| Project Status |
Completed (Fiscal Year 1990)
|
| Budget Amount *help |
¥6,800,000 (Direct Cost: ¥6,800,000)
Fiscal Year 1990: ¥1,400,000 (Direct Cost: ¥1,400,000)
Fiscal Year 1989: ¥5,400,000 (Direct Cost: ¥5,400,000)
|
|---|
| Keywords | cell culture / development / brain / sprouting / nerve cells / regeneration / cryostat section / cell attachment |
|---|
| Research Abstract |
Chick neocortical cells were cultured on cryostat tissue sections of the brain. Cells preferentially attached to the gray matter of adult rat CNS tissues. In contrast, they attached to any part of the brain when cultured on developing rat or marure frog brain tissues. Transection of fiber bundles at the superior cerebellar peduncle decussation of adult rat, which reportedly causes regeneration of cerebellofugal axons, made nearby white matter permissive to cell attachment. Superimposition of the gray matter of one section onto the white matter of another converted the former into a nonpermissive substrate for cell attachment, evidence suggesting that preferential cell attachment to the gray matter of abult rat CNS is due to inhibitory factor (s) localized in the white matter. This inhibitory factor appears to be absent in frog brain and developing rat brain.
To clarify the molecular mechanisms subserving the inhibition of cell attachment, we attempted to characterize and purify the factor from adult rat CNS. The inhibitory activity was solubilized into cholate/deoxycholate solution following extraction into a mixture of chroloform/methanol. This factor required reconstitution into liposomes for expression of its activity. By gel filtration (Superose 12), the inhibitory activity peaked at 25-45 kD and this peak of activity coincided with a peak of 280 nm absorbance. When the fractions including the peak activity were analyzed by SDS-polyacrylamide electrophoresis, several bands were detected. Thin layer chromatography also revealed lipids, possibiy phosphatidyl -choline and -ethanolamine. Hence the factor might be a complex of strongly-hydrophobic protein (s) and lipid (s) with relatively low molecular weight.
|
