| Project/Area Number |
14570157
|
|---|
| Research Category |
Grant-in-Aid for Scientific Research (C)
|
| Allocation Type | Single-year Grants |
|---|
| Section | 一般 |
|---|
| Research Field |
Human pathology
|
|---|
| Research Institution | Dokkyo Medical University |
|---|
Principal Investigator |
UEDA Yoshihiko Dokkyo University, School of Medicine, Professor, 医学部, 教授 (50151808)
|
|---|
| Project Period (FY) |
2002 – 2004
|
| Project Status |
Completed (Fiscal Year 2004)
|
| Budget Amount *help |
¥3,600,000 (Direct Cost: ¥3,600,000)
Fiscal Year 2004: ¥500,000 (Direct Cost: ¥500,000)
Fiscal Year 2003: ¥500,000 (Direct Cost: ¥500,000)
Fiscal Year 2002: ¥2,600,000 (Direct Cost: ¥2,600,000)
|
|---|
| Keywords | glomerular sclerosis / cytokine / tight junction / ZO-1 / microdissection / サイトカン / Cadherin |
|---|
| Research Abstract |
1.We conducted a clinicopathological studies of renal biopsy cases including segmental sclerotic lesion.
We classified renal disease according to the WHO classification system and investigated for proteinuria, hematuria, hypertension and others.
2.We investigated staining of cytokines and the localization of mRNA by in situ hybridization in biopsy specimens. By the immunostain TNF demonstrated extensive location in patients with Lupus nephritis as well as deposition in GBM.
3.The signals of cytokines, especially TNF, were extensively located in the mesangium in patients with Lupus nephritis, MPGN, membranous GN and some of diffuse mesangial PGN, and interstitium in minor glomerular abnormalities.
4.There was a different pattern of localization of staining and mRNA expression of cytokine (TNF) in patients with Lupus nephritis.
5.We investigated the expression of ZO-1 and phosphotyrosine in the patients with Lupus nephritis. Glomeruli demonstrated a positive stain consistent with localization of ZO-1 and Phosphotyrosine in the junction of foot process and slit diaphragm. We confirmed ZO-1 expression of glomeruli with Lupus nephritis. We hypothesized that the changes of glomerular permeability in Lupus nephritis was not associated with loss of ZO-1 induced to ZO-1 expression by phosphorylation.
6.We determined the protocol for laser capture microdissection and DNA extraction.
20μ.m thick specimens were cut in cryostat, and then were mounted onto a clear polyethylene membrane that was attached to an aluminum frame slide. To obtain only epithelial crypts for DNA extraction, we conducted laser capture microdissection. Microdissection was performed using a UV laser microdissection system and dissected crypts were collected on individual plastic caps. The caps were placed in a microcentrifuge tube, and the DNA was extracted using a DNA isolator PS kit according to the manufacture's protocol.
|
