Nofima Mat AS, Ås, Norway.
Fish Physiol Biochem. 2011 Dec;37(4):821-31. doi: 10.1007/s10695-011-9480-5. Epub 2011 Mar 31.
We have previously characterized the development of vertebral fusions induced by elevated water temperature in Atlantic salmon. Molecular markers of bone and cartilage development together with histology were used to understand the complex pathology and mechanism in the development of this spinal malformation. In this study, we wanted to use proteomics, a non-hypothetical approach to screen for possible new markers involved in the fusion process. Proteins extracted from non-deformed and fused vertebrae of Atlantic salmon were therefore compared by two-dimensional electrophoresis (2DE) and MALDI-TOF analysis. Data analysis of protein spots in the 2DE gels demonstrated matrilin-1, also named cartilage matrix protein, to be the most highly up-regulated protein in fused compared with non-deformed vertebrae. Furthermore, real-time PCR analysis showed strong up-regulation of matrilin-1 mRNA in fused vertebrae. Immunohistochemistry demonstrated induced matrilin-1 expression in trans-differentiating cells undergoing a metaplastic shift toward chondrocytes in fusing vertebrae, whereas abundant expression was demonstrated in cartilaginous tissue and chordocytes of both non-deformed and fused vertebrae. These results identifies matrilin-1 as a new interesting candidate in the fusion process, and ratify the use of proteomic as a valuable technique to screen for markers involved in vertebral pathogenesis.
我们之前已经描述了高温诱导大西洋鲑鱼椎体融合的发展过程。利用骨和软骨发育的分子标记物和组织学方法,我们试图了解这种脊柱畸形发生的复杂病理学和机制。在这项研究中,我们想使用蛋白质组学这一非假设性方法来筛选可能参与融合过程的新标记物。因此,我们通过二维电泳(2DE)和 MALDI-TOF 分析比较了来自未变形和融合的大西洋鲑鱼椎体的蛋白质提取物。2DE 凝胶中蛋白质斑点的数据分析表明,与未变形的椎体相比,聚集蛋白-1(也称为软骨基质蛋白)在融合的椎体中表达上调最明显。此外,实时 PCR 分析显示融合椎体中 matrilin-1 mRNA 强烈上调。免疫组织化学显示,在融合的椎体中,发生向软骨细胞的过渡性分化的细胞中诱导了 matrilin-1 的表达,而在未变形和融合的椎体的软骨组织和脊索细胞中均有大量表达。这些结果表明 matrilin-1 是融合过程中的一个新的候选标志物,并且证实了蛋白质组学作为一种筛选与椎体发病机制相关标志物的有价值技术的应用。