| Budget Amount *help |
¥4,160,000 (Direct Cost: ¥3,200,000、Indirect Cost: ¥960,000)
Fiscal Year 2009: ¥1,690,000 (Direct Cost: ¥1,300,000、Indirect Cost: ¥390,000)
Fiscal Year 2008: ¥2,470,000 (Direct Cost: ¥1,900,000、Indirect Cost: ¥570,000)
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| Research Abstract |
In the present study, diacylglycerol kinase (DGK) expression in rat aortic cells was investigated. In rat aortic smooth muscle cells (RASMC), α, ε and ζ types of DGK isozymes mRNA were detected. Immunocytochemical analysis revealed that DGKα, ε, ζ were localized in nucleus and cytoplasm, cytoskelton and nucleus, respectively. Phalloidin staining demonstrated that DGK_ε was localized to actin stress fibers. When RASMC were treated with serotonin, a contractile agonist, subcellular localization of DGKε was altered to be diffusely in cytoplasm. These facts indicate that DGK_ε functions in the regulatory mechanism of contraction in RASMC. In rat aortic endothelial cells (RAEC), immunocytochemical analysis revealed that DGK_γ was localized in Golgi complex. Present study demonstrated that up-regulated expression of DGK_γ mRNA by IL-1β. We also investigated the expression of cyclooxygenase-2 (COX-2) which was induced by IL-1β. In endothelial denuded aortae, COX-2 expression was induced in tunica media, in addition to adventitia, whereas no expression was detected in tunica media of intact aortae. These facts can be significant to study functional role of DGK isozymes, for example regulation of TRP channels, in vascular cells.
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