|Budget Amount *help
¥3,100,000 (Direct Cost : ¥3,100,000)
Fiscal Year 2000 : ¥700,000 (Direct Cost : ¥700,000)
Fiscal Year 1999 : ¥800,000 (Direct Cost : ¥800,000)
Fiscal Year 1998 : ¥1,600,000 (Direct Cost : ¥1,600,000)
The research which explores the cause of a small eye or a cataract is important for the development the methods for prevention and treatment, as well as for understanding the lens development system. This is the purpose of this research project. In this research, we have tried to know how mutated γE^crystallin caused the abnormal eye lens formation and to find a new physiological meaning for γE^crystallin. Further, lens filensin, which forms "bead-like filament" structure with CP49 and α-crystallin, is considered to be important for maintenance of lens transparency as well as lens formation. As no mutation for filensin genen have reported yet in neither human nor animals, it is recently found the hereditary cataract caused by mutated CP49 was reported, suggesting the filensin gane mutation also will cause similarly the cataract. There are many primary factors causing small eye or cataract, but in order to systematically understand the development of symptoms, it seemed significant to u
nderstand the expression mechanism of a filensin gene.
(1) Plasmid pEGFP-γ Elo involving mutated γE-crystallin cDNA was developed in cooperation with Dr. Quinlan in England. When the vector was introduced into the cultured lens cells amyloids were formed in the lens.
(2) We found a filensin gene promoter in the 5'-upstream region of a filensin gene, next the shortest region was defined as the promoter "an activity core." When the reporter plasmid, which put the "activity core" into the upstream of a luciferase gene, was transfected into the cell, the activity was shown in the non-lens cells, suggesting the existence of other active fragment for filensin gene lens specific expression. The fragment S12 (1.7kb) was found at about 6.5 Kb upstream region. The fragment S12 activated SV40 promoter and the HSV-TK promoter irrespective of the direction and position of insertion. The base sequence of S12 was determined and compared to human corresponding sequence, it was clear that these identity is intentionally high. Less