Cloning and functional analysis of Cl-ATPase 51kDa subunit
| Project/Area Number |
14580753
|
|---|
| Research Category |
Grant-in-Aid for Scientific Research (C)
|
| Allocation Type | Single-year Grants |
|---|
| Section | 一般 |
|---|
| Research Field |
Neurochemistry/Neuropharmacology
|
|---|
| Research Institution | Kansai Medical University |
|---|
Principal Investigator |
HATTORI Naoki Kansai Medical University, Faculty of Medicine, Associate Professor, 医学部, 助教授 (80288828)
|
|---|
| Project Period (FY) |
2002 – 2003
|
| Project Status |
Completed (Fiscal Year 2003)
|
| Budget Amount *help |
¥3,600,000 (Direct Cost: ¥3,600,000)
Fiscal Year 2003: ¥1,400,000 (Direct Cost: ¥1,400,000)
Fiscal Year 2002: ¥2,200,000 (Direct Cost: ¥2,200,000)
|
|---|
| Keywords | chloridw / ATPase / amyloid β / Alzheimer's disease / phosphatidylinositol-4-phosphate / Cl^--ATPase / イノシトール / グルタミン酸 / PI / PIP4 |
|---|
| Research Abstract |
Cl-ATPase enables the inhibitory neurotransmission by GABA by keeping the intracellular chloride concentration l* we previously reported that Cl-ATPase activity decreased in Alzheimer's disease brains and that low dose amyloid β(Aβ) inhibited Cl-ATPase activity leading to the elevation of intracellular chloride levels. In. the pres* study, (1)we performed the cloning of Cl-ATPase 51kDa subunit and (2)we examined the effects phosphatidylinositol 4 phosphate on Ap-induced inhibition of Cl-ATPase.
(1)We introduced reactive red column for the purification of Cl-ATPase to increase the recovery of the protein and set up sensitive assay for Cl-ATPase activity. We have identified several candidate proteins by mass spectrometric analysis on prot* spots separated two-dimensional electrophoresis.
(2)Low dose(10nM) of Aβ treatment for 2 days decreased Cl-ATPase activity by 47%, while did not affect Na/K-ATPase * anion insentive ATPase activities. Phosphatidylinositol(PI 50-750nM) and phosphatidylinositol-4-phosphate do* dependency recovered the Cl-ATPase activity that was suppressed by Aβ. Intracellular Cl concentration that was increa* three times by Aβ returned to normal by adding 75nM PI or PI4P. Low dose of Aβ(10nM) plus 10μM glutam* administration to neurons induced DNA fragmentation and reduced cell viability as shown by increased LDL release and WST assay. Seventy-five nM PI or PI4P recovered these cell damages. It is widely accepted that glutamate neurotoxicity involved in the pathogenesis of Alzheimer's disease. We demonstrated that elevation of intracellular chloride concentration * to the reduction of Cl-ATPase was possibly involved in the pathogenesis of neurodegeneration in Alzheimer's disease, * addition, we suggested that inositol might be used for the treatment or prevention of the progression of neurodegeneraton Alzheimer's disease.
|
Report
(3 results)
Research Products
(17 results)
