| Project/Area Number |
09671491
|
|---|
| Research Category |
Grant-in-Aid for Scientific Research (C)
|
| Allocation Type | Single-year Grants |
|---|
| Section | 一般 |
|---|
| Research Field |
Orthopaedic surgery
|
|---|
| Research Institution | Osaka University |
|---|
Principal Investigator |
NAKATA Ken Osaka University Medical School, Assistant Professor, 医学系研究科, 助手 (00283747)
|
|---|
| Co-Investigator(Kenkyū-buntansha) |
NAKASE Takanobu Osaka University Medical School, Assistant Professor, 医学系研究科, 助手 (00283755)
YASUI Natsuo Osaka University Medical School, Associate Professor, 医学系研究科, 助教授 (00157984)
|
|---|
| Project Period (FY) |
1997 – 1998
|
| Project Status |
Completed (Fiscal Year 1998)
|
| Budget Amount *help |
¥3,000,000 (Direct Cost: ¥3,000,000)
Fiscal Year 1998: ¥600,000 (Direct Cost: ¥600,000)
Fiscal Year 1997: ¥2,400,000 (Direct Cost: ¥2,400,000)
|
|---|
| Keywords | cartilage differentiation / transcription factor / tissue-specific transcription(control) / 軟骨 / 転写 / II型コラーゲン / 分化 / 組織特異的転写因子 |
|---|
| Research Abstract |
1. cDNA screening for candidate gene of tissue-specific transcription factors of cartilage
We have screened rhondrocyte cDNA expression library by South-western technique to identify cDNA clones which can interact with enhancer elements of type II collagen gene. Four positive clones (CSEBP ; cartilage specific enhancer binding protein) were further analyzed for their mRNA expressioin pattern by Nothern blotting and/or RT-PCR analysis. These clones were expressed in cartilage ant other tissues such as brain and hears in mouse embryos. One of these clones were cDNA for type II collagen C-propeptide.
2. Analysis of transgenic mouse overexpressing type II collagen C-propeptide.
In order to educidate in vivo function of type II collagen C-propeptide which was isolated as a binding protein of type II collagen gene enhancer, we have generated transgenic mice harboritig cDNA for type II collagen C-propeptide with type II collagen promoter/enhancer to achieve high level expression of this construc
… More
t in cartilage tissue These mice showed mild dwarfism with shortening of long bone and delayed calcification. Immunohistochemical analysis and immuno-electron microscopy demonstrated the expression of this construct in the extracelular matrix of hypertrophic cartilage in growth plate and in rER in hypersrophic chondrocyte. Furthermore, these transgene molecule was detectable in nucleus of hypertrophic cartilage.Taken together with our previous in vitro study, these results indicate the possibility that type II collagen C-propeptide was taken up hypertrophic chondrocyte, translocated to nucleus and bound to the enhancer region of type II collagen gene enhancer to reduce transcription of this gene as a negative feedback regulator.
3. Analysis of gene structure and production of knock-out mice of CSEBP
To analyze in vivo function of CSEBPs, we have screened mouse genomic library and constructed knock-out vector for one of these clones. CSEBP-2 has 14 exons expanding 8 Kb and this gene was expressed in ES cells. We have generated promoterless construct to generate knock-out mice for CSEBP-2. Less
|
