The purpose of this study is to evaluate the eligibility of gene therapy in treating glaucoma by constructing a replication deficient recombinant adenovirus carrying cDNA of stromelysin and by observing its expression in the rat eye. Stromelysin cDNA was obtained by RT-PCR using total RNA extracted from the cultured human trabecular cells. The cDNA was cloned into TA vector and subsequently into p deltaA. CMV. pA. The adenovirus vector containing stromelysin cDNA was constructed by co-transfection of pJM17 and p delta A. CMV-str. pA into the 293 cells. The expression of stromelysin in the trabecular meshwork and uveoscleral outflow channel of the rat eyes was detected by in situ hybridization and immunohistochemistry. In conclusion, this study showed the possibility of gene therapy in glaucoma by showing the expression of stromelysin with recombinant adenovirus vector containing stromelysin cDNA in the rat eye.
Adenoviridae* ; Animals ; Clone Cells ; DNA, Complementary ; Gene Expression* ; Genetic Therapy ; Glaucoma ; Humans ; Immunohistochemistry ; In Situ Hybridization ; Matrix Metalloproteinase 3* ; Rats* ; RNA ; Trabecular Meshwork

The purpose of this study is to establish the criteria to discriminate whether the difference of the optic disc parameters compared actually reflects the progress of the disease or it is merely caused by the intraexaminer or interexaminer error. Using the confocal scanning laser ophthalmoscope (TopSS), one eye of each of 20 healthy adults were exmained by the same examiner and one eye of each of 10 adults were examined by different examiners for two consecutive days. The results out of these repeated examinations were analysed to obtain 95%confidence intervals by analysis of the change of the 12 optic disc parameters, and to determine whether the results have been influenced by the disc size or the disc tilt. The criteria to differentiate actual progression of disease from potential errors of the test were obtained. In addition, it was confirmed that changes in the parameters were not influenced by the disc size or the contour tilt. In conclusion,the outcomes of this study can be utilized as a guiding criteria when judging the relevance of the parameter changes to the progress of the disease.
Adult ; Humans ; Ophthalmoscopes