Matrix metalloproteinases MMP2 and MMP9 are produced in early stages of kidney morphogenesis but only MMP9 is required for renal organogenesis in vitro.
Matrix metalloproteinases MMP2 and MMP9 are produced in early stages of kidney morphogenesis but only MMP9 is required for renal organogenesis in vitro.
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DOI：
10.1083/jcb.136.6.1363
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发表时间：
1997-03-24
期刊：
影响因子：
--
通讯作者：
Ronco PM
中科院分区：
文献类型：
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作者：
Lelongt B;Trugnan G;Murphy G;Ronco PM
我们分析了基质金属蛋白酶（MMP）的生产11天胚胎小鼠肾脏和这些酶对随后的肾器官发生的影响。在体内，免疫定位的金属蛋白酶的激光扫描共聚焦显微镜和肾裂解物的酶谱表明，胚胎肾间充质合成MMP 9和MMP 2酶。在体外，胚胎肾脏也分泌这两种酶时，培养在培养基中没有激素，生长因子和血清为24小时，在此期间出现T形分支的输尿管芽。然后，我们通过将抗MMP 2或抗MMP 9 IgG加入11-d肾脏的培养液中24或72 h来评估MMP 2和MMP 9在肾脏形态发生中的作用。虽然它抑制小鼠酶的活性，但抗MMP 2 IgG对肾脏形态发生没有影响。相反，具有酶阻断活性的抗MMP 9 IgG通过抑制输尿管芽的T形分支和进一步分裂，以浓度依赖性方式损害肾形态发生。这种作用是不可逆的，诱导事件后仍观察到，并通过外源性金属蛋白酶组织抑制剂1（TIMP 1），MMP 9的天然抑制剂再现。这些数据提供了第一次证明MMP 9和MMP 2在体内生产的11-D胚胎肾脏，并进一步表明，MMP 9是所需的在体外的分支形态发生的输尿管芽。
We analyzed matrix metalloproteinase (MMP) production by 11-d embryonic mouse kidneys and the effects of these enzymes on subsequent renal organogenesis. In vivo, immunolocalization of metalloproteinases by laser scanning confocal microscopy and zymograms of kidney lysates showed that the mesenchyme of embryonic kidneys synthesized both MMP9 and MMP2 enzymes. In vitro, embryonic kidneys also secreted both enzymes when cultured in a medium devoid of hormone, growth factor, and serum for 24 h during which T-shaped branching of the ureter bud appeared. We then evaluated the role of MMP2 and MMP9 in kidney morphogenesis by adding anti-MMP2 or anti-MMP9 IgGs to the culture medium of 11-d kidneys for 24 or 72 h. Although it inhibited activity of the mouse enzyme, anti-MMP2 IgGs had no effect on kidney morphogenesis. In contrast, anti-MMP9 IgGs with enzyme-blocking activity impaired renal morphogenesis, in a concentration-dependent manner, by inhibiting T-shaped branching and further divisions of the ureter bud. This effect was irreversible, still observed after inductive events and reproduced by exogenous tissue inhibitor of metalloproteinase 1 (TIMP1), the natural inhibitor of MMP9. These data provide the first demonstration of MMP9 and MMP2 production in vivo by 11-d embryonic kidneys and further show that MMP9 is required in vitro for branching morphogenesis of the ureter bud.