| Project/Area Number |
17590235
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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 |
General medical chemistry
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| Research Institution | Tokyo Medical and Dental University |
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Principal Investigator |
ABE Mayumi Tokyo Medical and Dental University, Graduate school academic staff of endowed department, 寄附講座教員 (80271980)
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| Co-Investigator(Kenkyū-buntansha) |
SATO Yasufumi Tohoku University Institute of Development, Aging and Cancer, Professor (50178779)
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| Project Period (FY) |
2005 – 2007
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| Project Status |
Completed (Fiscal Year 2007)
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| Budget Amount *help |
¥3,860,000 (Direct Cost: ¥3,500,000、Indirect Cost: ¥360,000)
Fiscal Year 2007: ¥1,560,000 (Direct Cost: ¥1,200,000、Indirect Cost: ¥360,000)
Fiscal Year 2006: ¥1,200,000 (Direct Cost: ¥1,200,000)
Fiscal Year 2005: ¥1,100,000 (Direct Cost: ¥1,100,000)
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| Keywords | vasculogenesis / angiogenesis / vascular endothelial cells / ES cells / vasohibin / PILSAP / differentiation / stem cells |
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| Research Abstract |
We examined the role of puromycin insensitive leucyl-specific aminopeptidase (PILSAP) in vasculogenesis. by using an embryoid bodies (Ebs) culture system. Flk-1 showed two expression peaks, days 4 and 10. These two peaks represent mesodermal precursors and mature Ecs, respectively. ES cells transfected with mutant PILSAP (mtPILSAP) cDNA of a dominant negative activity organized less vascular structure and showed decreased levels of endothelial markers. The similar results were obtained with an inhibitor of leucine aminopeptidase or siRNA. However, Flk-1 expression was unaffected on day 4. The expressions of markers for hematopoietic lineage and muscle cells in mtPILSAP-Ebs were lowered as well. These results suggest that although PILSAP may not function in the initial generation of Flk-1 positive mesodermal precursors, it dose play a role in growth of vascular, hematopoietic, and muscular lineage population from those precursors.
To clarify the mechanism how PILSAP is involved in differ
… More
entiation and/or proliferation of endothelial lineage, we performed proteome analysis using Mock-EB and mtPILSAP-EB of day 8 when other non-endothelial lineage cells are loosing Flk-1. Among the nuclear proteins whose expression was significantly decreased in mtPILSAP-EB, we focused our attention on Pigpen. Pigpen is one of the proteins in cajal/coiled body and has been reported to be expressed in growing and tumor vascular endothelial cells and angioblasts in embryo. However, the precise function of Pigpen is largely unknown. Inhibition of Pigpen expression in MG1.19 using siRNA, the expressions of markers for endothelia, hematopoietic, and muscle cell lineages were lowered as well as the case of PILSAP siRNA. Moreover, PILSAP and Pigpen can be associated in nucleus or on nuclear membrane and the binding of these molecules was enhanced by VEGF. Furthermore, this association was suppressed in mtPILSAP-EB. These data suggest that PILSAP and Pigpen cooperate in differentiation of endothelial lineage and vascular network formation. Less
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