| Budget Amount *help |
¥3,600,000 (Direct Cost: ¥3,600,000)
Fiscal Year 2003: ¥500,000 (Direct Cost: ¥500,000)
Fiscal Year 2002: ¥2,000,000 (Direct Cost: ¥2,000,000)
Fiscal Year 2001: ¥1,100,000 (Direct Cost: ¥1,100,000)
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| Research Abstract |
The expression levels of Growth arrest and DNA damage inducible gene (GADD) 153 are very low in normal growing cells and are highly induced in response to a variety of cellular stresses, including glucose deprivation, exposure to alkylating agents, oxidative stress, and growth-arresting situations. We previously reported that GADD153 is highly induced by oxidative stress, PPAR-gamma activator in vascular smooth muscle cells (VSMC). Further over-expression of GADD153 gene induced cell growth and apoptosis. In this study, to evaluate the GADD153 gene function in vivo, we generated transgenic rats over-expressing GADD153 through a mouse smooth muscle alpha-actin (SMA) promoter. The SMA-GADD153 chimeric gene was constructed by fusing a 3.6-kb fragment of the mouse SMA promoter to an entire coding region of rat GADD153. Total four times injection were performed, however only 4 SMA-GADD153 founder rats, which were identified by polymerase chain reaction, were obtained. We compared GADD153 gene and protein expression in the aortas between transgenic and non-transgenic rat by Northern and Western blotting. Expression levels of GADD153 gene and protein were almost same between them. On the same time, we generated SMA-CCAAT/enhancer binding protein-delta (CEBPD) transgenic rat In this case, only one injection. we obtained 8 SMA-CEBPD transgenic rats, which highly expressed CEBPD gene and protein in the aortas. These results and in vitro function of GADD153 suggest that SMA-GADD153 transgenic rat might be lethal in the embryo. We could not get the gadd153 transgenic rat, however. this study is useful to understand the in vivo function of GADD153 gene.
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