Unique expression patterns of matrix metalloproteinases in regenerating newt limbs
Unique expression patterns of matrix metalloproteinases in regenerating newt limbs
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DOI：
10.1002/dvdy.10247
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发表时间：
2003-02-01
影响因子：
2.5
通讯作者：
Yoshizato, K
中科院分区：
文献类型：
--
作者：
Kato, T;Miyazaki, K;Yoshizato, K
下肢再生的过程包括由基质金属蛋白酶（MMPs）诱导的细胞外基质（ecm）的剧烈重塑，这被认为是再生的触发因素之一。本研究以日本蝾螈MMPs （nMMPs）的5个基因（nMMP9、nMMP3/10-a、nMMP3/10-b、nMMP13和本研究新克隆的nMMPe）为探针，研究了日本蝾螈四肢的这种重塑。nMMPe全长502个氨基酸残基，与其他已知的脊椎动物MMPs具有较低的同源性。逆转录聚合酶链反应分析表明，nMMPe的转录本定位于顶端表皮帽（AEC）和非胚芽创面表皮，而非胚芽间充质和正常表皮。Northern blot分析发现，nMMP9、nMMP3/10-a和nMMP13转录本定位于再生肢体骨，而nMMP3/10-b转录本定位于AEC骨。mRNA原位杂交实验鉴定了表达nmmp的细胞。nMMP9基因在骨骺软骨软骨细胞中强烈表达。有趣的是，AEC的基底细胞，而非正常皮肤的基底细胞，强烈表达nMMP3/10-b。免疫组化分析显示，软骨细胞合成的nMMP9蛋白在截肢面骨基底膜和ecm中广泛分泌和分布。本研究中描述的这些nMMPs可能共同作用于重塑再生肢体的ecm。(C) 2003 Wiley-Liss, Inc。
The process of regeneration of urodele limbs includes a drastic remodeling of extracellular matrices (ECMs) that is induced by matrix metalloproteinases (MMPs) and is thought to be one of the triggers of the regeneration. We studied this remodeling in limbs of Japanese newt, Cynops pyrrhogaster, by using five genes of newt MMPs (nMMPs) as probes: nMMP9, nMMP3/10-a, nMMP3/10-b, and nMMP13 that had been characterized previously, and nMMPe that was newly cloned in the present study. nMMPe was 502 amino acid residues long and showed a low homology to other known vertebrate MMPs. Reverse transcriptase-polymerase chain reactions analysis localized the transcript of nMMPe in the apical epidermal cap (AEC) and the non-blastemal wound epidermis but not in the blastemal mesenchyme or the normal epidermis. Northern blot analysis localized the transcripts of nMMP9, nMMP3/10-a, and nMMP13 in the bone of regenerating limbs, whereas those of nMMP3/10-b in AEC. mRNA in situ hybridization experiments identified the nMMP-expressing cells. nMMP9 gene was strongly expressed in chondrocytes of the cartilage of epiphysis. Of interest, basal cells of AEC, but not those of the normal skin, expressed nMMP3/10-b intensely. Immunohistochemical analysis showed that the nMMP9 proteins synthesized by chondrocytes were secreted and distributed widely in the basement membrane of bone and ECMs of the amputation plane. These nMMPs characterized in the present study might cooperatively work to remodel ECMs of regenerating limbs. (C) 2003 Wiley-Liss, Inc.