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Analysis of the lymphatic development using mutant mice

使用突变小鼠分析淋巴发育

基本信息

批准号：
12670294
负责人：
YOSHIDA Nobuaki
金额：
$ 2.05万
依托单位：
The University of Tokyo
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
2000
资助国家：
日本
起止时间：
2000 至 2001
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-12670294/
关键词：
lymphatic vessels mutant mouse lymphatic edema transgenic mouse LYVE-1 マウス lyve-1

项目摘要

The lymphatics are thought to be responsible for edematous condition in patients, especially in patients suffering from primary lymphedema. Resent studies show that lymphangiogenesis, as well as angiogenesis, also plays some roles on tumor metastasis. However, the lymphatic development in mammals has been unknown from lack of reliable anatomical / histological procedure for the detection of lymphatic vassals and useful mutant animal that has obvious lymphatic abnormality.In order to understand the mechanism of lymphatic development and functions, we are generating "reporter" strains by transgenesis or targeted mutagenesis, using marker genes (lacZ, GFP, PLAP) under transcriptional regulation of the lymphatic endothelial-specific genes, such as LYVE-1, prox-1 and VFGFR-3. We have cloned mouse LYVE-1 gene and determined the genomic structure and chromosomal location of the gene in mice. For the construction of the transgenes using Prox-1 gene or VEGFR-3 gene, we purchased and re-analyzed their P1 or BAC clones. We are now introducing some constructs into mouse fertilized eggs. We have also started on the conditional knockout analyses of potential growth factor / receptor and transcriptional factors in lymphangiogenesis.We are also under investigation of an original spontaneous mutant mouse line developing chylous ascites, intestinal lymphedema and edematous hindfoot that are thought to be due to lymphatic abnormality. The blood flow is found not only in blood vessels but also in lymphatic vessels in the homozygous mutants. According to the confocal microscopic analysis of TRITC-conjigated gelatin injected into blood vessels, the peripheral capillary-lacteal shunt at the intestinal villi is one of the cause for blood flow observed lymphatics of the homozygous mutant mice. We are trying forward genetic approaches to find the candidate gene (s) for this mutation.

这些药物被认为是导致患者水肿的原因，尤其是原发性水肿患者。近年来的研究表明，淋巴管生成与血管生成一样，在肿瘤转移中也起一定的作用。然而，由于缺乏可靠的解剖学/组织学方法来检测淋巴管的发育，以及缺乏有用的突变动物来检测淋巴管的发育和功能，我们正在利用标记基因通过转基因或靶向突变来产生“报告”菌株，以了解淋巴管发育和功能的机制在淋巴管内皮特异性基因如LYVE-1、prox-1和VFGFR-3的转录调控下，这些基因可被转录激活（lacZ、GFP、PLAP）。我们克隆了小鼠LYVE-1基因，并确定了该基因在小鼠中的基因组结构和染色体定位。为了使用Prox-1基因或VEGFR-3基因构建转基因，我们购买并重新分析了它们的P1或BAC克隆。我们现在正在将一些构建体引入小鼠受精卵中。我们还开始了对淋巴管生成中潜在的生长因子/受体和转录因子的条件性敲除分析。我们还在研究一种原始的自发突变小鼠系，这种小鼠系出现乳糜腹水、肠水肿和后足水肿，这些被认为是由于淋巴管异常。在纯合突变体中，不仅在血管中发现血流，而且在淋巴管中也发现血流。根据注射到血管中的TRITC-缀合的明胶的共聚焦显微镜分析，肠绒毛处的外周毛细血管-乳管分流是纯合突变小鼠的血液流变学观察到的原因之一。我们正在尝试遗传学方法来寻找这种突变的候选基因。