Roles of the MFH-1 gene and WT 1 gene in kidney development

MFH-1 基因和 WT 1 基因在肾脏发育中的作用

基本信息

  • 批准号:
    08044238
  • 负责人:
  • 金额:
    $ 1.02万
  • 依托单位:
  • 依托单位国家:
    日本
  • 项目类别:
    Grant-in-Aid for international Scientific Research
  • 财政年份:
    1996
  • 资助国家:
    日本
  • 起止时间:
    1996 至 无数据
  • 项目状态:
    已结题

项目摘要

We isolated the MFH-1 gene as one of the forkhead gene family in 1993. The family are growing and more than 20 genes are known in mice. The MFH-1 gene is expressed at the somitic mesoderm and later expressed strongly in the sclerotome. It is also expressed in the developing metanephros and later on in the glomerulus after birth.We made knockout mice of the MFH-1 gene by homologous recombination in the ES cells. First, we cloned the mouse genomic MFH-1 gene and determined its genomic structure. We found that it has no introns. We made a targeting vector by replacing the exon with neo-resistance cassette and introduced it into ES cells by electroporation. Out of 480 neomycin- and gancyclovir-resistant clones, 6 were identified to be homologous recombinants. We made chimera mice by injecting them into a blastocyst and obtained heterozygotic mice by mating chimera mice and wild B6 mice. The heterozygotes were asymptomatic. However, homozygotes were dead within ten minutes after birth.Homozygotes had abnormalities in several tissues. Here we focused the phenotype of kidneys. The size of kidneys in homozygotes was half that of the wild-type. Histologically the glomerulus, renal tubular cells and blood vesssels were present normally. But dilated renal calces were seen in homozygotes.Next, we cloned the human genomic MFH-1 gene and determined its nucleotide sequence. We found that human MFH-1 protein is 501 amino acids long and the amino acid sequence of the forkheah domain is identical in human and mouse. Identity of the mouse and human MFH-1 protein is 94%.We also made monoclonal antibodies against the human MFH-1 protein. Using thses antibodies, we found that the MFH-1 protein is expressed in chondrosarcoma, osteosarcoma and Wilms tumor cell lines.
1993年,我们分离了叉头基因家族中的MFH-1基因。这个家族正在壮大,在小鼠中已知的基因超过20个。MFH-1基因在体节中胚层表达,随后在硬节中强烈表达。MFH-1基因在胚胎干细胞中的同源重组法构建了MFH-1基因敲除小鼠模型。首先,我们克隆了小鼠基因组MFH-1基因,并确定了其基因组结构。我们发现它没有内含子。我们将新耐药基因外显子替换成靶向载体,并通过电穿孔法将其导入ES细胞。在480个新霉素和更昔洛韦抗性克隆中,6个被鉴定为同源重组体。我们通过将它们注射到胚泡中来制造嵌合体小鼠,并通过嵌合体小鼠和野生B6小鼠交配来获得杂合子小鼠。杂合子无症状。然而,纯合子在出生后10分钟内死亡。纯合子在几个组织中有异常。在这里,我们重点关注肾脏的表型。纯合子的肾脏大小是野生型的一半。组织学上肾小球、肾小管细胞和血管正常。其次,我们克隆了人基因组MFH-1基因,并测定了其核苷酸序列。我们发现,人MFH-1蛋白的长度为501个氨基酸,并且叉头结构域的氨基酸序列在人和小鼠中是相同的。小鼠MFH-1蛋白与人MFH-1蛋白的同源性为94%。利用这些抗体,我们发现MFH-1蛋白在软骨肉瘤、骨肉瘤和Wilms肿瘤细胞系中表达。

项目成果

期刊论文数量(15)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)
Hirasawa,F.: "The effect of silver administration on the biosynthesis and molecular propartics of rat ceruloplasmin" Biochim.Biophys.Acta. (in press).
Hirasawa,F.:“银给药对大鼠铜蓝蛋白生物合成和分子原配体的影响”Biochim.Biophys.Acta。
  • DOI:
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    0
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Masuda Y.: "Platelet-derived growth factor B-chain homodimer suppressing a tonic-clonic convulsion of seizure-prone E1 mice" Biochim.Biophys.Res.Commun.233. 60-63 (1996)
Masuda Y.:“血小板衍生生长因子 B 链同二聚体抑制癫痫易发 E1 小鼠的强直阵挛惊厥”Biochim.Biophys.Res.Commun.233。
  • DOI:
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    0
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  • 通讯作者:
Masuda,Y.: "Platelet-derived growth factor B-chain homodimer suppressing a tonic-clonic convulsion fo seizure-prone El mouse." Biochim. Biophys. Res. Commun.223. 60-63 (1996)
Masuda,Y.:“血小板衍生生长因子 B 链同二聚体抑制癫痫易发性 El 小鼠的强直阵挛性惊厥。”
  • DOI:
  • 发表时间:
  • 期刊:
  • 影响因子:
    0
  • 作者:
  • 通讯作者:
Miura,N.: "Isolation of the mouse and human MFH-1 (Mesenchyme Fork Head-1) genes reveals conscrvation of their gene and protein structures." Genomics. (in press).
Miura,N.:“小鼠和人类 MFH-1(间充质叉头 1)基因的分离揭示了它们基因和蛋白质结构的保守性。”
  • DOI:
  • 发表时间:
  • 期刊:
  • 影响因子:
    0
  • 作者:
  • 通讯作者:
Masuda,Y: "Platclet-derived growth factor B-chain homodimer suppressing a tonic-clomic convulsion of scizure pronc El mouse." Biochim.Biophys.Res.Commun.223. 60-63 (1996)
Masuda,Y:“血小板源性生长因子 B 链同二聚体抑制 scizure pronc El 小鼠的强直性痉挛。”
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  • 影响因子:
    0
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MIURA Naoyuki其他文献

MIURA Naoyuki的其他文献

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{{ truncateString('MIURA Naoyuki', 18)}}的其他基金

Generation of the HCV-infectable mouse-an animal model for inflammation cancer
HCV感染小鼠的产生——炎症性癌症动物模型
  • 批准号:
    24659603
  • 财政年份:
    2012
  • 资助金额:
    $ 1.02万
  • 项目类别:
    Grant-in-Aid for Challenging Exploratory Research
Generation of mice in which the mouse hepatocytes are replaced with the human hepaocytes and its application
人肝细胞替代小鼠肝细胞的小鼠产生及其应用
  • 批准号:
    19390347
  • 财政年份:
    2007
  • 资助金额:
    $ 1.02万
  • 项目类别:
    Grant-in-Aid for Scientific Research (B)
Molecular mechanism of hepatic carcinogenesis and hepatocyte apoptosis in the Rb transgenic mice
Rb转基因小鼠肝癌发生及肝细胞凋亡的分子机制
  • 批准号:
    15390393
  • 财政年份:
    2003
  • 资助金额:
    $ 1.02万
  • 项目类别:
    Grant-in-Aid for Scientific Research (B)
Molecular machanism of the MFH-1 gene in, the aoortic arch formation, Skeletogenesis and kidney formation
MFH-1基因在主动脉弓形成、骨骼发生和肾脏形成中的分子机制
  • 批准号:
    11694239
  • 财政年份:
    1999
  • 资助金额:
    $ 1.02万
  • 项目类别:
    Grant-in-Aid for Scientific Research (B)
Molecular investigation on the hepatic caicinogenesis-resistant model animals
肝脏抗癌模型动物的分子研究
  • 批准号:
    11557090
  • 财政年份:
    1999
  • 资助金额:
    $ 1.02万
  • 项目类别:
    Grant-in-Aid for Scientific Research (B)
Molecular investigation on the fulminant hepatitis-resistant model animals
暴发性肝炎耐药模型动物的分子研究
  • 批准号:
    10470253
  • 财政年份:
    1998
  • 资助金额:
    $ 1.02万
  • 项目类别:
    Grant-in-Aid for Scientific Research (B).
ROLES OF THE MFH-1 AND WT1 GENES IN KIDNEY DEVELOPMENT
MFH-1 和 WT1 基因在肾脏发育中的作用
  • 批准号:
    09044252
  • 财政年份:
    1997
  • 资助金额:
    $ 1.02万
  • 项目类别:
    Grant-in-Aid for international Scientific Research
GENERATION AND APPLICATION OF THE MODEL MICE WHICH SHOW RESISTANCE TO RENAL CARCINOGENESIS
抗肾癌小鼠模型的产生及应用
  • 批准号:
    08557089
  • 财政年份:
    1996
  • 资助金额:
    $ 1.02万
  • 项目类别:
    Grant-in-Aid for Scientific Research (A)
Lsolation and characterization of the Brain Forkhead gene
脑叉头基因的分离和表征
  • 批准号:
    05680592
  • 财政年份:
    1993
  • 资助金额:
    $ 1.02万
  • 项目类别:
    Grant-in-Aid for General Scientific Research (C)
Molecular Cloning of Rat Hepatocyte Nuclear Factor 1 Gene and Analysis of Its Promotor
大鼠肝细胞核因子1基因的分子克隆及其启动子分析
  • 批准号:
    02670108
  • 财政年份:
    1990
  • 资助金额:
    $ 1.02万
  • 项目类别:
    Grant-in-Aid for General Scientific Research (C)

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阐明金属硫蛋白基因敲除小鼠的寿命与肉碱生物合成能力之间的关系。
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