Normal newt limb regeneration requires matrix metalloproteinase function
Normal newt limb regeneration requires matrix metalloproteinase function
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DOI：
10.1016/j.ydbio.2004.12.003
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发表时间：
2005-03-01
影响因子：
2.7
通讯作者：
Odelberg, SJ
中科院分区：
文献类型：
--
作者：
Vinarsky, V;Atkinson, DL;Odelberg, SJ
蝾螈通过一个复杂的过程再生失去的肢体，包括去分化、迁移、增殖和截肢平面附近细胞的再分化。为了确定控制这些细胞事件的基因，我们进行了再生和nonregenerating从蝾螈Notophthalmus viridescens肢体之间的差异显示分析。这项分析，再加上直接克隆的方法，确定了一个以前未知的Notophthalmus胶原酶基因（nCol）和三个已知的基质金属蛋白酶（MMP）基因，MMP 3/10 a，MMP 3/10 b和MMP 9，所有这些都是在截肢数小时内上调。MMP 3/10 b表现出最高和最普遍的表达，并且似乎占肢体中蛋白水解活性的大部分，如通过酶谱法测量的。通过测试纯化的重组MMP蛋白对潜在的底物，我们表明，nCol是一个真正的胶原酶，MMP 9是明胶酶，MMP 3/10 a是一个溶基质，MMP 3/10 b有一个异常广泛的基板配置文件，作为溶基质和非经典的胶原酶。将再生肢体暴露于合成的MMP抑制剂GM 6001会产生矮小、畸形的肢体再生或具有远端瘢痕的肢体残端。这些数据表明，MMPs是正常蝾螈肢体再生所必需的，MMPs的功能部分是防止再生过程中的瘢痕形成。(C)2004年爱思唯尔公司All rights reserved.
Newts regenerate lost limbs through a complex process involving dedifferentiation, migration, proliferation, and redifferentiation of cells proximal to the amputation plane. To identify the genes controlling these cellular events, we performed a differential display analysis between regenerating and nonregenerating limbs from the newt Notophthalmus viridescens. This analysis, coupled with a direct cloning approach, identified a previously unknown Notophthalmus collagenase gene (nCol) and three known matrix metalloproteinase (MMP) genes, MMP3/10a, MMP3/10b, and MMP9, all of which are upregulated within hours of limb amputation. MMP3/10b exhibits the highest and most ubiquitous expression and appears to account for the majority of the proteolytic activity in the limb as measured by get zymography. By testing purified recombinant MMP proteins against potential substrates, we show that nCol is a true collagenase, MMP9 is a gelatinase, MMP3/10a is a stromelysin, and MMP3/10b has an unusually broad substrate profile, acting both as a stromelysin and noncanonical collagenase. Exposure of regenerating limbs to the synthetic MMP inhibitor GM6001 produces either dwarfed, malformed limb regenerates or limb stumps with distal scars. These data suggest that MMPs are required for normal newt limb regeneration and that MMPs function, in part, to prevent scar formation during the regenerative process. (C) 2004 Elsevier Inc. All rights reserved.