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Research of the physiological functions of membrane-type matrix metalloproteinases

膜型基质金属蛋白酶的生理功能研究

基本信息

批准号：
13470050
负责人：
SEIKI Motoharu
金额：
$ 10.05万
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 (B)
财政年份：
2001
资助国家：
日本
起止时间：
2001 至 2002
项目状态：
已结题

项目摘要

In our murine strain deficient in MT1-MMP gene expression, the bone morphogenesis was severely impaired, consistent with the past reports. Next question would be an address to know which cell lineages in bone tissue are responsible to work with expressing MT1-MMP during the bone morphogenesis under the physiologic state. To see if a recovering of the MT1-MMP limitedly in bone tissue helps to revart the phenotype, a conditional transgenic strain is under construction based on the knock-out mice, in which expression of MT1-MMP is recovered restrictedly in osteoblasts. On the other hand, MT1-MMP plays a important role in angiogenesis around the ossification center during the second ossification. In order to understand the role more profoundly, three-dimensional culture of muscle chunks in type-I collagen has been conducted. Angiogenesis through the collagen was observed during the culturing period. The angiogenesis was severely impaired with the mucle chunk of the MT1-MMP deficient strain … More compared to the wild type or the hetero-zygote.MT1-MMP in the new vessels was predominantly expressed in endothelial cells. Questions remains how MT1-MMP plays a role in angiogenesis, why other proteinases can not compensate the functions sufficiently, or whether the impairment in second ossification in MT1-MMP deficient mice is induced predominantly by the poor angiogenesis around the bone. The conditional transgenic mice could provide us with some hints to ask for the queries.MT5-MMP has been found to expressed at neural cells such in cerebrum, cerebellum, and hypocampus. However, no distinct abnormality has been detected in a strain deficient in MT5-MMP gene. Based on the specificity in MT5-MMP expression, a neurite extension assay was performed with the murine neural cells derived from dorsal route ganglia, demonstrated that the protrusion of the neurite was facilitated on laminin, and inhibited on heparin binding proteoglycan. Adding a soluble MT5-MMP purified, The latter inhibition has released with a degradation of the substrate.Immunohistochemically, endogenous MT5-MMP was detected predominantly at the growth cone.MT4-MMP has been shown to be expressed such at a cortex in the frontal lobe of brain, hypocampus, cerebellum, nephron, smooth muscle cells in the vessels, heart, and red pulp in spleen. Histologically, no gloss abnormality was found in a strain- deficient in MT4-MMP gene. Several functional analyzes should be performed next to know how MT4-MMP plays roles in murine tissue. Less

在我们的 MT1-MMP 基因表达缺陷的小鼠品系中，骨形态发生严重受损，与过去的报道一致。下一个问题是了解生理状态下骨形态发生过程中骨组织中哪些细胞谱系负责表达 MT1-MMP。为了了解 MT1-MMP 在骨组织中的有限恢复是否有助于恢复表型，我们正在基于敲除小鼠构建条件转基因菌株，其中 MT1-MMP 的表达在成骨细胞中受到限制的恢复。另一方面，MT1-MMP在第二次骨化过程中骨化中心周围的血管生成中发挥重要作用。为了更深入地了解其作用，对I型胶原蛋白中的肌肉块进行了三维培养。在培养期间观察到通过胶原的血管生成。与野生型或杂合子相比，MT1-MMP 缺陷株的粘液块的血管生成严重受损。新血管中的 MT1-MMP 主要在内皮细胞中表达。问题仍然是 MT1-MMP 如何在血管生成中发挥作用，为什么其他蛋白酶不能充分补偿其功能，或者 MT1-MMP 缺陷小鼠的第二次骨化受损是否主要是由骨周围血管生成不良引起的。条件转基因小鼠可以为我们提出疑问提供一些线索。MT5-MMP已被发现在大脑、小脑和海丘等神经细胞中表达。然而，在MT5-MMP基因缺陷的菌株中没有检测到明显的异常。基于MT5-MMP表达的特异性，用源自背路神经节的小鼠神经细胞进行神经突延伸测定，证明层粘连蛋白促进神经突的突出，而肝素结合蛋白聚糖抑制神经突的突出。添加纯化的可溶性MT5-MMP，后者的抑制随着底物的降解而释放。免疫组织化学，内源性MT5-MMP主要在生长锥中检测到。MT4-MMP已被证明在大脑额叶皮层、海下丘、小脑、肾单位、血管平滑肌细胞等处表达，心，红髓在脾。组织学上，MT4-MMP基因缺陷菌株未发现光泽异常。接下来应该进行一些功能分析，以了解 MT4-MMP 如何在小鼠组织中发挥作用。较少的