Large-scale analysis of the genes involved in fin regeneration and blastema formation in the medaka, Oryzias latipes
Large-scale analysis of the genes involved in fin regeneration and blastema formation in the medaka, Oryzias latipes
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DOI：
10.1016/j.mod.2004.03.015
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发表时间：
2004-07-01
影响因子：
2.6
通讯作者：
Kudo, A
中科院分区：
文献类型：
--
作者：
Katogi, R;Nakatani, Y;Kudo, A
青竹是研究表型再生的一个很有吸引力的模型。鳍具有显著的再生能力，截肢后约14天可更换。胚泡是一组未分化的细胞，它的形成是再生所必需的；然而，其分子机制还不完全清楚。为了确定鳍再生所需的基因，特别是胚泡的形成，我们构建了截肢后3天和10天的鳍再生的cDNA文库。共测序了16,866个表达序列标签(EST)，并进行了BLASTX分析。结果表明，其中约60%的蛋白质与已鉴定的蛋白质具有较强的匹配性，并分离出了与发育相关的主要信号分子，包括成纤维细胞生长因子、骨形态发生蛋白、Wnt、Notch/Delta和EPhin/Eph信号通路。为了寻找在鳍再生过程中特异表达的新基因，从每个与已知蛋白质序列没有相似性的文库中筛选出2900个独立的EST，构建了基因芯片。通过竞争杂交和原位杂交的基因表达谱筛选，获得了6个与胚泡形成相关的候选基因。Olrfe16d23和olrfe14k04仅在胚泡形成的早期再生阶段表达。Olrf5n23编码一种新的信号肽，在创面再生的整个过程中都有表达。Olrfe23122、olrfe20n22和olrfe24i02在胚胎区有显著表达。因此，我们的研究确定了基因表达谱和一些新的候选基因，以便于阐明鳍再生的分子机制。(C)2004爱思唯尔爱尔兰有限公司。保留所有权利。
Medaka is an attractive model to study epimorphic regeneration. The fins have remarkable regenerative capacity and are replaced about 14 days after amputation. The formation of blastema, a mass of undifferentiated cells, is essential for regeneration; however, the molecular mechanisms are incompletely defined. To identify the genes required for fin regeneration, especially for blastema formation, we constructed cDNA libraries from fin regenerates at 3 days postamputation and 10 days postamputation. A total of 16,866 expression sequence tags (ESTs) were sequenced and subjected to BLASTX analysis. The result revealed that about 60% of them showed strong matches to previously identified proteins, and major signaling molecules related to development, including FGF, BMP, Wnt, Notch/Delta, and Ephrin/Eph signaling pathways were isolated. To identify novel genes that showed specific expression during fin regeneration, cDNA microarray was generated based on 2900 independent ESTs from each library which had no sequence similarity to known proteins. We obtained 6 candidate genes associated with blastema formation by gene expression pattern screening in competitive hybridization analyses and in situ hybridization. Olrfe16d23 and olrfe14k04 were expressed only in early regenerating stages when blastema formation was induced. The expression of olrf5n23, which encodes a novel signal peptide, was detected in wound epidermis throughout regeneration. Olrfe23122, olrfe20n22 and olrfe24i02 were expressed notably in the blastema region. Our study has thus identified the gene expression profiles and some novel candidate genes to facilitate elucidation of the molecular mechanisms of fin regeneration. (C) 2004 Elsevier Ireland Ltd. All rights reserved.