| Project/Area Number |
15590906
|
|---|
| Research Category |
Grant-in-Aid for Scientific Research (C)
|
| Allocation Type | Single-year Grants |
|---|
| Section | 一般 |
|---|
| Research Field |
Neurology
|
|---|
| Research Institution | Dokkyo Medical University |
|---|
Principal Investigator |
MURAKAMI Nobuyuki Dokkyo Medical School, School of Medicine, Department of Pediatrics, Lecturer, 医学部, 講師 (00316598)
|
|---|
| Co-Investigator(Kenkyū-buntansha) |
SAKUTA Ryoichi Dokkyo Medical School, School of Medicine, Department of Pediatrics, Associate Professor, 医学部, 助教授 (40254974)
|
|---|
| Project Period (FY) |
2003 – 2005
|
| Project Status |
Completed (Fiscal Year 2005)
|
| Budget Amount *help |
¥3,500,000 (Direct Cost: ¥3,500,000)
Fiscal Year 2005: ¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 2004: ¥1,000,000 (Direct Cost: ¥1,000,000)
Fiscal Year 2003: ¥1,500,000 (Direct Cost: ¥1,500,000)
|
|---|
| Keywords | regenerating muscle / TGF-beta 2 / plasmid / direct gene transfer / RNA interference / プラスミド / βガラクトシダーゼ / 筋 |
|---|
| Research Abstract |
To investigate the function of TGF-beta 2 in regenerating fiber, we injected pBR322 plasmid attached with LacZ and LTR in mouse regenerating muscle into which we injected bupivacain before 72 hours. On 7days after plasmid injection, we took the regenerating mouse muscles. Frozen sections were β-galactosidase stained. However, we could not find any β-galactosidase positive muscle fibers in the regenerating fibers. To check how the plasmid works in normal mouse muscle, we injected 10μg, 50μg, 100μg of the plasmid in normal mouse muscle. We found a few β-galactosidase positive muscle fibers even in 100μg injected normal mouse muscles. We changed pBR322 plasmid vector into pCMVbeta one. We performed the same experiment using pCMVbeta vector. We could find β-galactosidase positive muscle fibers in the muscles that 50μg and 100μg of the plasmid were injected. We made plasmid attached with sense or anti-sense of TGF-beta 2 and LacZ. We injected 50μg of the plasmid into normal mouse muscles. On 7 days after the plasmid injection, we sacrificed mice and the muscles were immediately frozen in isopentane cooled with liquid nitrogen. Frozen sections were β-galactosidase stained and immunostained with anti-TGF beta 2 antibodies. However we could not find β-galactosidase or TGF-beta 2 positive muscle fibers. TGF beta 2 gene is not so long. Gene size might be a factor that prevents gene transfection of TGF beta 2.
|
