Expression of junctional adhesion molecule-1 in rat corneal tissue
10.3760/cma.j.issn.2095-0160.2012.03.003
- VernacularTitle:接合黏附分子-1在大鼠角膜中的表达研究
- Author:
Guang-jie, WANG
;
Hua, JIANG
- Publication Type:Journal Article
- Keywords:
Junctional adhesion molecule-1;
Tight junction;
Cornea;
Reverse transcription polymerase chain reaction;
hnmunohistochemistry
- From:
Chinese Journal of Experimental Ophthalmology
2012;30(3):204-208
- CountryChina
- Language:Chinese
-
Abstract:
BackgroundJunctional adhesion molecule-1 (JAM-1) is intercellular transmembrane protein newly discovered and associated with the tight junction.Tight junction plays an important role in keeping the transparency of cornea,but there are few studies about JAM-1 in cornea tight junction.ObjectiveThis study was to determine the expression of JAM-1 in corneal epithelium,stroma,endothelium,and locate the distribution of JAM-1.MethodsThe corneas of two SFP Wistar rats were obtained,and the samples of epithelial lamella,corneal stroma and endothelium with Descemet membrane were prepared respectively for the detection of the expression of JAM-1 mRNA using reverse transcription polymerase chain reaction(RT-PCR).Primers were designed according to the genes as RGD web provided,and the objective genes were JAM-1,occludin and claudin-1.Products of PCR were examined by 1.5％ agarose gel electrophoresis and assayed with GelDoc-lt UVP Imaging System.The corneal paraffin sections and stretched preparations of epithelium and endothelium of corneal sections from other two SFP Wistar rats were prepared for the examination of JAM-1protein expression and location by immunohistochemistry.The use of experimental animals followed the Statement of ARVO. Results JAM-1,occludin and claudin-1 mRNA were expressed in rat cornea epithelium,stroma and endothelium.PCR melting curves showed the limpid unimedality.The expression level of JAM-1 mRNA was similar to occludin mRNA and higher than that of claudin-1 with the highest level in the epithelium layer.Immunohistochemistry assay revealed that there was definite JAM-1 antibody staining in the cornea epithelium,stroma and endothelium layers.However,the basal layer of corneal epithelium presented with the strongest staining in comparison with stromal and endothelial layers.Stretched preparations of corneal epithelium and endothelium showed that JAM-1 protein appeared at the junction site of epithelial cells and endothelial cells.The basal layer of the corneal epithelium showed the strongest response,and the staining of corneal endothelium was extensive and diffuse. Conclusions JAM-1 is a composition of intercellular tight junction which expresses in cornea epithelium,endothelium and stroma.However,its appearance and level vary from different corneal layers.
