García Dana M, Bauer Hermann, Dietz Thomas, Schubert Thomas, Markl Jürgen, Schaffeld Michael
Department of Biology, Texas State University-San Marcos, San Marcos, TX 78666, USA.
Cell Tissue Res. 2005 Nov;322(2):245-56. doi: 10.1007/s00441-005-0031-1. Epub 2005 Nov 3.
With more than 50 genes in human, keratins make up a large gene family, but the evolutionary pressure leading to their diversity remains largely unclear. Nevertheless, this diversity offers a means to examine the evolutionary relationships among organisms that express keratins. Here, we report the analysis of keratins expressed in two cyprinid fishes, goldfish and carp, by two-dimensional polyacrylamide gel electrophoresis, complementary keratin blot binding assay, and immunoblotting. We further explore the expression of keratins by immunofluorescence microscopy. Comparison is made with the keratin expression and catalogs of zebrafish and rainbow trout. The keratins among these fishes exhibit a similar range of molecular weights and isoelectric points, with a similar overall pattern on two-dimensional gels. In addition, immunofluorescence microscopy studies of goldfish and carp tissues have revealed the expression of keratins in both epithelial and mesenchymally derived tissues, as reported previously for zebrafish and trout. We conclude that keratin expression is qualitatively similar among these fishes, with goldfish and carp patterns being more similar to each other than to zebrafish, and the cyprinid fishes being more similar to each other than to the salmonid trout. Because of the detected similarity of keratin expression among the cyprinid fishes, we propose that, for certain experiments, they are interchangeable. Although the zebrafish distinguishes itself as being a developmental and genetic/genomic model organism, we have found that the goldfish, in particular, is a more suitable model for both biochemical and histological studies of the cytoskeleton, especially since goldfish cytoskeletal preparations seem to be more resistant to degradation than those from carp or zebrafish.
人类中有50多个基因编码角蛋白，它们构成了一个庞大的基因家族，但导致其多样性的进化压力在很大程度上仍不清楚。然而，这种多样性为研究表达角蛋白的生物体之间的进化关系提供了一种方法。在这里，我们报告了通过二维聚丙烯酰胺凝胶电泳、互补角蛋白印迹结合分析和免疫印迹法对角蛋白在两种鲤科鱼类金鱼和鲤鱼中表达情况的分析。我们还通过免疫荧光显微镜进一步探究了角蛋白的表达。并与斑马鱼和虹鳟鱼的角蛋白表达及分类进行了比较。这些鱼类中的角蛋白表现出相似的分子量范围和等电点，在二维凝胶上具有相似的整体模式。此外，金鱼和鲤鱼组织的免疫荧光显微镜研究显示，角蛋白在上皮组织和间充质来源的组织中均有表达，这与之前报道的斑马鱼和鳟鱼情况一致。我们得出结论，这些鱼类之间角蛋白的表达在性质上相似，金鱼和鲤鱼的模式彼此之间比与斑马鱼的模式更相似，鲤科鱼类彼此之间比与鲑科鳟鱼更相似。由于在鲤科鱼类中检测到角蛋白表达的相似性，我们建议在某些实验中它们可以相互替代。尽管斑马鱼作为一种发育和遗传/基因组模式生物具有独特地位，但我们发现，特别是金鱼，对于细胞骨架的生化和组织学研究是更合适的模型，尤其是因为金鱼的细胞骨架制剂似乎比鲤鱼或斑马鱼的制剂更耐降解。
