| Project/Area Number |
15591752
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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 |
Obstetrics and gynecology
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| Research Institution | HIROSHIMA UNIVERSITY |
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Principal Investigator |
KUDO Yoshiki Hiroshima University, Graduate School of Biomedical Sciences, Professor, 大学院・医歯薬学総合研究科, 教授 (80241082)
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| Project Period (FY) |
2003 – 2005
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| Project Status |
Completed (Fiscal Year 2005)
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| Budget Amount *help |
¥3,200,000 (Direct Cost: ¥3,200,000)
Fiscal Year 2005: ¥900,000 (Direct Cost: ¥900,000)
Fiscal Year 2004: ¥900,000 (Direct Cost: ¥900,000)
Fiscal Year 2003: ¥1,400,000 (Direct Cost: ¥1,400,000)
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| Keywords | Placenta / Trophoblast / Syncytialisation / Cell fusion / Amino acid transport / Pre-eclampsia |
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| Research Abstract |
(1)A unique biological process of placental trophoblast syncytialisation has been studied using a cell model of syncytialisation (the cytotrophoblast cell line BeWo following increased intracellular cAMP by forskolin).
(2)Stable BeWo cell lines constitutively expressing either a fusion protein green fluorescent protein and human histone H2B or a fusion protein of red fluorescent protein and the mitochondrial targeting sequence from subunit VIII of human cytochrome c oxidase have been generated. Using these two BeWo cell lines expressing fluorescent protein of different colour cell fusion (syncytialisation) has been visualized and a fluorescence activated cell sorting based assay for determining quantitatively the rate of syncytialisation has been developed.
(3)The time course of syncytialisation has been studied using DNA microarray in a cell model of syncytialisation. Among the genes with increased expression following forskolin treatment were many required for cellular communication and metabolism. Several genes known to be involved in cell adhesion and fusion have markedly changed expression levels very early following forskolin exposure, thus preceding morphological fusion of BeWo cells. Further analysis of this data and expression profiling in general will be able to contribute to understanding the functional basis for the formation of the placental syncytiotrophoblast.
(4)The physiological importance of CD98 surface antigen in regulating syncytialisation and amino acid transport activity has been studied in parallel using either antisense oligonucleotides or RNA interference. The results show that CD98 is involved in the process of cell fusion necessary for syncytiotrophoblast formation and that during this physiologically important event amino acid transport activity is also regulated through expression of this membrane protein.
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