| Project/Area Number |
10557011
|
|---|
| Research Category |
Grant-in-Aid for Scientific Research (B)
|
| Allocation Type | Single-year Grants |
|---|
| Section | 展開研究 |
|---|
| Research Field |
General medical chemistry
|
|---|
| Research Institution | KYOTO UNIVERSITY |
|---|
Principal Investigator |
TSUKITA Shoichiro KYOTO UNIVERISTY, FACULTY of MEDICINE PROFESSOR, 医学研究科, 教授 (50155347)
|
|---|
| Co-Investigator(Kenkyū-buntansha) |
ITOH Masahiko KYOTO UNIVERSTIY, FACULTY of MEDICINE ASSISTANT, 医学研究科, 助手 (70270486)
FURUSE Mikio KYOTO UNIVERSTIY, FACULTY of MEDICINE ASSISTANT, 医学研究科, 助手 (90281089)
TSUKITA Sachiko KYOTO UNIVERSITY COLLEGE of MEDICAL TECHNOLOGY PROFESSOR, 医療技術短期大学部, 教授 (00188517)
|
|---|
| Project Period (FY) |
1998 – 1999
|
| Project Status |
Completed (Fiscal Year 1999)
|
| Budget Amount *help |
¥12,200,000 (Direct Cost: ¥12,200,000)
Fiscal Year 1999: ¥5,400,000 (Direct Cost: ¥5,400,000)
Fiscal Year 1998: ¥6,800,000 (Direct Cost: ¥6,800,000)
|
|---|
| Keywords | occludin / tight junction / embryonic stem cell / ZO-1 / embryoid body / paracellular pathway / drug delivery / barrier / ノックアウト / ドミナントネガティブ / 内皮細胞 / 阻害ペプチド |
|---|
| Research Abstract |
Occludin is the only known integral membrane protein of tight junctions (TJ), and is now believed to be directly involved in the barrier and fence functions of TJ. Occludin-deficient embryonic stem (ES) cells were generated by targeted disruption of both alleles of the occludin gene. When these cells were subjected to suspension culture, they aggregated to form simple, and then cystic embryoid bodies (EBs) with the same time course as EB formation from wild-type ES cells. Immunofluorescence microscopy and ultrathin section electron microscopy revealed that polarized epithelial (visceral endoderm-like) cells were differentiated to delineate EBs not only from wild-type but also from occludin-deficient ES cells. Freeze fracture analyses indicated no significant differences in number or morphology of TJ strands between wild-type and occludin-deficient epithelial cells. Furthermore, ZO-1, a TJ-associated peripheral membrane protein, was still exclusively concentrated at TJ in occludin-deficient epithelial cells. In good agreement with these morphological observations, TJ in occludin-deficient epithelial cells functioned as a primary barrier to the diffusion of a low molecular mass tracer through the paracellular pathway. These findings indicate that there are as yet unidentified TJ integral membrane protein (s) which can form strand structures, recruit ZO-1, and function as barrier without occludin.
|
