Max-Planck-Institute for Heart and Lung Research, Parkstr, 1, 61231 Bad Nauheim, Germany.
BMC Genomics. 2010 Jan 4;11:4. doi: 10.1186/1471-2164-11-4.
The newt Notophthalmus viridescens possesses the remarkable ability to respond to cardiac damage by formation of new myocardial tissue. Surprisingly little is known about changes in gene activities that occur during the course of regeneration. To begin to decipher the molecular processes, that underlie restoration of functional cardiac tissue, we generated an EST database from regenerating newt hearts and compared the transcriptional profile of selected candidates with genes deregulated during zebrafish heart regeneration.
A cDNA library of 100,000 cDNA clones was generated from newt hearts 14 days after ventricular injury. Sequencing of 11520 cDNA clones resulted in 2894 assembled contigs. BLAST searches revealed 1695 sequences with potential homology to sequences from the NCBI database. BLAST searches to TrEMBL and Swiss-Prot databases assigned 1116 proteins to Gene Ontology terms. We also identified a relatively large set of 174 ORFs, which are likely to be unique for urodele amphibians. Expression analysis of newt-zebrafish homologues confirmed the deregulation of selected genes during heart regeneration. Sequences, BLAST results and GO annotations were visualized in a relational web based database followed by grouping of identified proteins into clusters of GO Terms. Comparison of data from regenerating zebrafish hearts identified biological processes, which were uniformly overrepresented during cardiac regeneration in newt and zebrafish.
We concluded that heart regeneration in newts and zebrafish led to the activation of similar sets of genes, which suggests that heart regeneration in both species might follow similar principles. The design of the newly established newt EST database allows identification of molecular pathways important for heart regeneration.
蝾螈具有通过形成新的心肌组织来应对心脏损伤的显著能力。令人惊讶的是，对于在再生过程中发生的基因活性变化，人们知之甚少。为了开始破译构成功能性心脏组织恢复的分子过程，我们从再生蝾螈心脏中生成了一个 EST 数据库，并将选定候选基因的转录谱与斑马鱼心脏再生过程中失调的基因进行了比较。
从心室损伤后 14 天的蝾螈心脏中生成了 100000 个 cDNA 克隆的 cDNA 文库。对 11520 个 cDNA 克隆进行测序，得到 2894 个组装的连续序列。BLAST 搜索显示，1695 个序列与 NCBI 数据库中的序列具有潜在同源性。BLAST 搜索到 TrEMBL 和 Swiss-Prot 数据库将 1116 个蛋白分配到 Gene Ontology 术语。我们还鉴定了一组相对较大的 174 个 ORF，它们可能是蝾螈两栖动物所特有的。新蛙-斑马鱼同源物的表达分析证实了心脏再生过程中某些基因的失调。序列、BLAST 结果和 GO 注释在基于关系的网络数据库中可视化，然后将鉴定的蛋白质分组到 GO 术语的簇中。比较再生斑马鱼心脏的数据，确定了在蝾螈和斑马鱼心脏再生过程中普遍过表达的生物学过程。
我们得出结论，蝾螈和斑马鱼的心脏再生导致了相似的基因集的激活，这表明这两个物种的心脏再生可能遵循相似的原则。新建立的蝾螈 EST 数据库的设计允许鉴定对心脏再生重要的分子途径。
