2000 Fiscal Year Final Research Report Summary
ANALYTICAL RESEARCH FOR THE MOLECULAR MECHANISM OF MOUSE SMALL EYE AND CATARACT.
| Project/Area Number |
10671664
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Ophthalmology
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| Research Institution | INSTITUTE FOR DEVELOPMENTAL RESEARCH |
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Principal Investigator |
MASAKI Shigeo Institute for Developmental Research, Aichi Human Service Center, Department of Biochemistry, Senior Researcher, 生化学部, 主任研究員 (10157175)
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| Co-Investigator(Kenkyū-buntansha) |
TAKEHANA Makoto Kyoritsu Pharmaceutical College, Department of Physiology and Anatomy, Lecturer., 生理解剖学, 講師 (60121505)
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| Project Period (FY) |
1998 – 2000
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| Keywords | cataract / γ-crystallin / filensin / cell differentiation / organ-specific expression / transcription factor / expression abnormality / lens |
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| Research Abstract |
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 gene have reported yet in neither human nor animals, it is recently found the hereditary cataract caused by mutated CP49 was reported, suggesting the filensin gene 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 seemsed significant to u
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nderstand the expression mechanism of a filensin gene.
(1) Plasmid pEGFP-γElo 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
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