| Project/Area Number |
06807137
|
|---|
| Research Category |
Grant-in-Aid for General Scientific Research (C)
|
| Allocation Type | Single-year Grants |
|---|
| Research Field |
Ophthalmology
|
|---|
| Research Institution | Juntendo University School of Medicine |
|---|
Principal Investigator |
UEDA Shunsuke Juntendo University School of Medicine, Assistant Professor, 医学部, 講師 (70129774)
|
|---|
| Project Period (FY) |
1994 – 1995
|
| Project Status |
Completed (Fiscal Year 1995)
|
| Budget Amount *help |
¥800,000 (Direct Cost: ¥800,000)
Fiscal Year 1995: ¥400,000 (Direct Cost: ¥400,000)
Fiscal Year 1994: ¥400,000 (Direct Cost: ¥400,000)
|
|---|
| Keywords | Rat cornea / sympathetic fibers / corneal nerve fibers / 角膜神経 / 定量的観察 / 神経線維密度 / 免疫組織化学的染色法 |
|---|
| Research Abstract |
The purpose of the study was to investigate the best procedure for corneal nerve density analysis after the wounding. Rat, rabbit and guinea pig corneal epithelia were subjected to the histological and immunohistochemical study. Free epithelial sheets were obtained by incubating the corneas in balanced salt solution with 20 mM ethyl-enediaminetetraacetate. The epithelial sheets were processed by the histological procedure (gold impregnation technique) or immunohitochemical procedure (avidin biotin complex method). Neuron specific enolase (NSE : nonspecific neural tissue marker), calcitonin gene related peptide (CGRP : sensory nerve marker) and tyrosine hydroxylase (TH : sympathtic nerve marker) were targeted by the immunohistochemical procedure.
The epithelial sheets stained by the gold impregnation almost always revealed the perfect profile of intraepithelial nerve fibers in all specimens examined. The quality of the staining was kept good enough for quantitative analysis for at least several weeks. The gold impregnated epithelial sheets seemed to be more useful for the quantitative study of the intraepithelial nerve than conventional procedures. The epithelial sheets stained by NSE and CGRP immunohistochemical rocedure revealed the similar nerve fiber pattern observed in the gold impregnated specimen, however the quality of staining was worse than that of the gold impregnated specimen. TH positive fibers which seemed to be sympathetic fibers were occasionally observed. The immunohistochemical procedure was incomplete for the quantitative analysis of intraepithelial nerve fibers currently.
|
