Matrix metalloproteinase expression and function during fin regeneration in zebrafish: Analysis of MT1-MMP, MMP2 and TIMP2
Matrix metalloproteinase expression and function during fin regeneration in zebrafish: Analysis of MT1-MMP, MMP2 and TIMP2
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DOI：
10.1016/j.matbio.2005.03.007
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发表时间：
2005-06-01
期刊：
影响因子：
6.9
通讯作者：
Sarras, MP
中科院分区：
文献类型：
--
作者：
Bai, S;Thummel, R;Sarras, MP
基质金属蛋白酶（MMPs）在细胞外基质（ECM）的周转中起关键作用，因此，在发育过程中作为细胞-ECM相互作用的关键调节剂发挥作用。尽管它们在发育过程中的重要性，相对较少的报道，这些金属蛋白酶在肢体发育和再生过程中的作用。为了解决肢体（鳍）再生过程中细胞-ECM相互作用的问题，我们利用斑马鱼作为实验模型。本研究在本实验室前期MMP基因克隆研究的基础上，对膜型金属蛋白酶（MT 1-MMP）、明胶酶A（MMP-2）和内源性金属蛋白酶组织抑制剂2（TIMP-2）在成年斑马鱼鳍再生过程中的表达进行了研究。原位分析表明，共表达zmt 1-mmp，zmmp-2和ztimp-2 mRNA转录在再生尾鳍。原位明胶酶谱证实了再生鳍中存在活性金属蛋白酶。在尾鳍再生过程中，zmt 1-mmp、zmmp-2和ztimp-2 mRNA转录物在胚基和基底上皮中表达，而IV型胶原[zcol-IV（a5）]转录物（基底层组分）的表达仅限于基底上皮。在GM 6001（MMP活性抑制剂）的存在下，鳍生长大大减少，表明这些酶在鳍再生过程中的重要性。Itch的先前研究（EMBO，2001）表明，仅含有血红素跨膜胞质结构域（MT 1HPX）的脊椎动物MT 1-MMP构建体的表达导致MT 1-MMP嗜同性复合物形成的阻断和随后的pro-MMP-2活化的抑制。干扰嗜同性复合物的形成归因于在细胞表面的血红素结合蛋白结构域的表达。在这些早期发现的基础上，目前的研究发现，鳍再生中MT 1HPX的异位表达抑制了再生过程，并导致芽基中细胞增殖的减少。综上所述，这些结果表明，MMPs在斑马鱼鳍再生过程中具有重要作用。(c)2005 Elsevier B.V/国际基质生物学学会。All rights reserved.
Matrix metalloproteinases (MMPs) play key roles in the turnover of extracellular matrix (ECM) and, thereby, function as key regulators of cell-ECM interactions during development. In spite of their importance during developmental processes, relatively little has been reported about the role of these metalloproteinases during limb development and regeneration. To approach the problem of cell-ECM interactions during limb (fin) regeneration, we have utilized zebrafish as an experimental model. Based on previous MMP cloning studies from our laboratory, the current study has focused on the expression of membrane-type I metalloproteinase (MT1-MMP), gelatinase A (MMP-2) and endogenous tissue inhibitor 2 of metalloproteinases (TIMP-2) during fin regeneration in adult zebrafish. In situ analysis indicated co-expression of zmt1-mmp, zmmp-2, and ztimp-2 mRNA transcripts in regenerating caudal fins. In situ gelatin-zymography confirmed the presence of active metalloproteinases in regenerating fins. zmt1-mmp, zmmp-2, and ztimp-2 mRNA transcripts were expressed in the blastema and basal epithelium during caudal fin regeneration while expression of type IV collagen [zcol-IV(a5)] transcripts (a basal lamina component) was restricted to the basal epithelium. Fin outgrowth was greatly reduced in the presence of GM6001 (an inhibitor of MMP activity) indicating the importance of these enzymes during fin regeneration. Previous studies by Itch (EMBO, 2001) indicated that expression of a vertebrate MT1-MMP construct containing only the hemopexin-transmembrane-cytoplasmic domains (MT1HPX) resulted in blockage of MT1-MMP homophilic complex formation and subsequent inhibition of pro-MMP-2 activation. Interference with homophilic complex formation was attributed to expression of the hemopexin domain at the cell surface. Building upon these earlier findings, the current study found that ectopic expression of MT1HPX in fin regenerates inhibited the regeneration process and resulted in a reduction in cell proliferation in the blastema. Taken together, these results indicate that MMPs have an important role during fin regeneration in zebrafish. (c) 2005 Elsevier B.V/Intemational Society of Matrix Biology. All rights reserved.