| Project/Area Number |
15300131
|
|---|
| Research Category |
Grant-in-Aid for Scientific Research (B)
|
| Allocation Type | Single-year Grants |
|---|
| Section | 一般 |
|---|
| Research Field |
Neurochemistry/Neuropharmacology
|
|---|
| Research Institution | TOKYO METROPOLITAN ORGANIZATION FOR MEDICAL RESEARCH |
|---|
Principal Investigator |
SHIBASAKI Futoshi TOKYO METROPOLITAN ORGANIZATION FOR MEDICAL RESEARCH, Tokyo Metropolitan Institute of Medical Science, 東京都臨床医学総合研究所, 副参事研究員 (90300954)
|
|---|
| Co-Investigator(Kenkyū-buntansha) |
KATO Hiroyuki TOKYO METROPOLITAN ORGANIZATION FOR MEDICAL RESEARCH, Tokyo Metropolitan Institute of Medical Science (30301732)
UCHINO Hiroyuki Tokyo Medical University, Hachioji Medical Center, 八王子医療センター・麻酔科, 講師 (60266476)
TAKENAWA Tadaomi The University of Tokyo, The Institute of Medical Science, 医科学研究所・癌・細胞増殖大部門・腫瘍分子医学研究部, 教授 (40101315)
|
|---|
| Project Period (FY) |
2003 – 2004
|
| Project Status |
Completed (Fiscal Year 2004)
|
| Budget Amount *help |
¥16,900,000 (Direct Cost: ¥16,900,000)
Fiscal Year 2004: ¥5,700,000 (Direct Cost: ¥5,700,000)
Fiscal Year 2003: ¥11,200,000 (Direct Cost: ¥11,200,000)
|
|---|
| Keywords | Brain Ischemia / Hypoxia / mouse ischemic model / calcineurin / calcium / siRNA / immunosuppressant / angiogenesis / 遅発性神経細胞死 / 虚血 / カルシウム受容体 / 遺伝子解析 / サイクロスポリンA / FK506 |
|---|
| Research Abstract |
We have reported that signal transduction pathway involving calcineurin and cydophilin D is a novel target for developing anti-ischemic drugs. For further analysis to clarify the role of both enzymes, we measured the intracellular calcium in pyramidal neuronal cells in hippocampal shoes after stimulation of hypoxia and low glucose with or without immunosuppressants cydosporin A or FK506. The results suggested that calcineurin activity might regulate the uptake of calcium through unknown calcium channel existed on plasma membranes.
In addition, we found the new relation between calcineurin and hypoxia responsive factor (HIF), which was reported to be critical in inducing factors in angiogenesis, glycolytic metabolism, apoptosis, and erythrocytosis. This hypoxic factors deeply involved in mechanisms by which hypoxia played an important role in ischemic brain damage and degenerative diseases such as Parkinson disease and Alzheimer disease. In our experiments, calcineurin can stabilize the
… More
expression and activate the transcription of HIFs. We performed yeast two hybrid to discover novel factors regulating HIF and found novel positive clones which suppressed HIF2 transcription activity as well as expression. One of fished done identified is Int6/eIF3e, which involed in breast cancer caused by mouse mammalian tumor virus (MMTV). MMTV was integrated into genome at several integration sites and damaged the target genes. Int6 was one of the target gene. Further analysis using recombinant Int6 mutants showed that Int6 was a tumor suppressor by direct binding to HIF2 and the degradation. We demonstrated the potent angiogenesis in mouse skin after injection of si RNA which specifically suppressed HIF2. Under these results, we started analysis using mice transgened with a Int6 dominant negative mutant or siRNA expression vector.
According to our specific aims in this research plan, we tried to establish a mouse ischemic model (forebrain mouse ischemic model). Two factors were critical One was the maintenance of body temperature. We equipped acryl bords surrounding microscope to keep 37℃. Another factor was selection of ischemic mouse after ligation of carotid arteries. Around 20 % of operated mice was observed to have enough blood supply through co-lateral blood flow by Doppler flow meter. It was regret that we could not evaluate PLC D1 KO mice nor NCX2 KO mice, but we plan to perform evaluation of Int6 siRNA or dominant negative Tg mice even after finishing this project. Less
|
