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Studies on the Pinctada fucata BMP-2 Gene: Structural Similarity and Functional Conservation of Its Osteogenic Potential within the Animal Kingdom

机译:Pinctada fucata BMP-2基因的研究:在动物界内其成骨潜力的结构相似性和功能保守性

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Bone morphogenetic protein (BMP) -2 plays an important role in morphogenesis in both vertebrates and invertebrates. BMP-2 is one of the most powerful bioactive substances known to induce the osteogenic differentiation of mesenchymal cells. We examinedthe structural and functional conservation of Pinctada fucata BMP-2 in inducing osteogenesis in the murine mesenchymal stem cells, C3H10T1/2. Exposure of C3H10T1/2 cells to the recombinant mature fragment of Pinctada fucata BMP-2 resulted in osteoblastic differentiation. The sequence, S VPKPCC VPTELSSL, within the C-terminal portion of Pinctada fucata BMP-2, is homologous to the knuckle epitope of human BMP-2. This synthetic polypeptide was able to induce differentiation of C3H10T1/2 along the osteoblastic lineage, as confirmed by an increase in alkaline phosphatase activity, and the accumulation of calcium, as determined by von Kossa staining. Furthermore, using immunohistochemical staining, we observed an increased expression of collagen type I, osteopontin, and osteocalcin, which are known markers of osteogenesis. These results show that BMP-2 is conserved, not only in terms of its homology at the amino acid sequence, but also in terms of driving the formation of hard tissues in vertebrates and invertebrates.
机译:骨形态发生蛋白(BMP)-2在脊椎动物和无脊椎动物的形态发生中都起着重要作用。 BMP-2是已知最强大的生物活性物质之一,可诱导间充质细胞的成骨分化。我们检查了Pinctada fucata BMP-2在诱导小鼠间充质干细胞C3H10T1 / 2中成骨的结构和功能保守性。 C3H10T1 / 2细胞暴露于Pinctada fucata BMP-2的重组成熟片段会导致成骨细胞分化。 Pinctada fucata BMP-2 C端部分的序列S VPKPCC VPTELSSL与人BMP-2的指关节表位同源。该合成的多肽能够诱导成骨细胞谱系的C3H10T1 / 2分化,这可以通过碱性磷酸酶活性的增加和钙的积累来证实,如von Kossa染色所确定的。此外,使用免疫组织化学染色,我们观察到了I型胶原蛋白,骨桥蛋白和骨钙素的表达增加,它们是成骨的已知标志。这些结果表明,BMP-2不仅在氨基酸序列上具有同源性,而且在驱动脊椎动物和无脊椎动物中硬组织形成方面均是保守的。

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