2000 Fiscal Year Final Research Report Summary
Analysis of the regulatory mechanism of the differentiation of epidermal keratinocytes mediated by fat-soluble ligands
| Project/Area Number |
10460117
|
|---|
| Research Category |
Grant-in-Aid for Scientific Research (B).
|
| Allocation Type | Single-year Grants |
|---|
| Section | 一般 |
|---|
| Research Field |
Zootechnical science/Grassland science
|
|---|
| Research Institution | TOKYO UNIVERSITY OF AGRICULTURE AND TECHNOLOGY |
|---|
Principal Investigator |
UEHARA Kohkichi Tokyo Univ. Agricul. & Technol. Dept. of Tissue Physiol. Prof., 農学部, 教授 (10014953)
|
|---|
| Co-Investigator(Kenkyū-buntansha) |
ARAI Koji Tokyo Univ. Agricul. & Technol. Dept. of Tissue Physiol. Assis. Prof., 農学部, 助手 (70293016)
ARAI Katsuhiko Tokyo Univ. Agricul. & Technol. Dept. of Vet Clinic. Assoc. Prof., 農学部, 助教授 (60175940)
MATSUDA Hiroshi Tokyo Univ. Agricul. & Technol. Dept. of Tissue Physiol. Prof., 農学部, 教授 (80145820)
|
|---|
| Project Period (FY) |
1998 – 2000
|
| Keywords | KERATINOCYTES / DIFFERENTIATION / KERATOHYALIN / MONOCLONAL ANTIBODY / 分化 |
|---|
| Research Abstract |
The monoclonal antibody reacted with keratohyalin granules was cloned. This antibody, MAbKH1 was generated by the immunization of cytoskeletal fraction prepared from canine mammary cacinoma against mice. MAbKH1 reacted the epidermis of several animal species including dog, rat, cattle and pig. In neonatal rat tissue, MAbKH1 also reacted with arterial and intestinal smooth muscle in addition of keratohyalin granules and in adult rat, this positive reaction was restricted in the epidermis immunohistochemically. Immunoblot analysis revealed that MAbKH1-recognizing protein in addition of 165 kDa and 200 kDa bands in both neonatal and adult rat skin extracts. The 1.7 kbp cDNA was cloned by the immunoscreening with MAbKH1 against lamda gt 11 constructed neonatal rat brain cDNA library. In vitro, expression of KH1 protein was upregulated under high calcium conditions accompanied with increased expression of transglutaminase, which is a molecular marker for the terminal differentiation of keratinocytes in rat keratinocytes. By double-labeled immunofluorescent staining with MAbKH1 and anti-cytokeratin rabbit serum, keratohyalin granules appeared by higher calcium ion were strongly stained with MAbKH1 in the cytoplasm of keratinocytes, suggesting that 55 kDa KH1 protein is one of constitution of keratohyalin granules. In conclusion, we characterized a new keratohyalin protein, which gave a molecular weight of 55 kDa and demonstrated that expression of this protein was regulated by the concentration of calcium ion in vitro.
|
