2004 Fiscal Year Final Research Report Summary
Molecular mechanisms for hemato-vascular formation
| Project/Area Number |
15390086
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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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 | University of Tsukuba |
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Principal Investigator |
YAMAMOTO Masayuki University of Tsukuba, Graduate School of Comprehensive Human Sciences, Professor, 大学院・人間総合科学研究科, 教授 (50166823)
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| Co-Investigator(Kenkyū-buntansha) |
SHIMIZU Ritsuko University of Tsukuba, Graduate School of Comprehensive Human Sciences, Lecturer, 大学院・人間総合科学研究科, 講師 (40226262)
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| Project Period (FY) |
2003 – 2004
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| Keywords | Hematopoietic stem cell / GATA-2 / GFP knock-in mouse in the GATA-2 locus / GATA-2 conditional knock-out mouse / Bone marrow transplantation |
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| Research Abstract |
We have established green fluorescent protein (GFP) knock-in knockout mice (G2-GFP), in which the GFP gene was inserted into Gata2 gene under the hematopoietic specific promoter/ first exon and analyzed the phenotypes of GFP expressing cells. The percentage of GFP positive cells was 0.5 % in the bone marrow mononuclear cells. These GFP positive cells, expressing c-Kit without any staining lineage markers (Lin), had a potential to develop several lineage committing cells on the OP9 cell layer, suggesting that GFP-positive cells were belonging to either the population of hematopoietic progenitor cells or that of hematopoietic stem cells. The expression of cellular proliferation-related molecules, such as c-Myc, Cyclin A2 and Cyclin B1, in the GFP-positive cells were higher than those in the GFP-negative cells. In addition, more than 30 % of the cells in the population of Lin-/c-Kit^+/GFP^+ accumulated in S/G2/M phase. These findings indicated that GATA-2 had an important role for the tra
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nscriptional regulation of genes relating to cell cycle progression of the immature hematopoietic progenitor cells.
To further investigate the function of GATA-2 on the hematopoietic cell proliferation in vivo, we established Gata2 gene conditional knockout allele (G2-CKO), in which GATA-2 cDNA sandwiched between two loxP sequences was inserted into the translation initiation site of Gata2 gene. Because the simple Gata2 gene knockout mice were embryonic lethal, this G2-CKO mouse line might be one of the most appropriate means to examine the function of GATA-2 on bone marrow hematopoiesis. We intercrossed the compound GATA-2 targeting mice (G2^<CKO/GFP>) with Mx1-Cre transgenic mice, in which the expression of Cre recombinase was induced by the treatment of interferon, and successfully generated the triple compound mouse colony (Gata2^<CKO/GFP>::Mx1-Cre). We are now ready for bone marrow transplantation analyses using the bone marrow cells of these mice to examine the function of GATA-2 for adult hematopoiesis. Less
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