Laboratory of Extracellular Matrix Biochemistry, Institute for Protein Research, Osaka University, 3-2 Yamadaoka, Suita, Osaka 565-0871, Japan.
Shimoda Marine Research Center, University of Tsukuba, 5-10-1 Shimoda, Shizuoka 415-0025, Japan.
Exp Anim. 2021 Aug 6;70(3):378-386. doi: 10.1538/expanim.21-0001. Epub 2021 Apr 6.
Sea urchin embryos have been one of model organisms to investigate cellular behaviors because of their simple cell composition and transparent body. They also give us an opportunity to investigate molecular functions of human proteins of interest that are conserved in sea urchin. Here we report that human disease-associated extracellular matrix orthologues ECM3 and QBRICK are necessary for mesenchymal cell migration during sea urchin embryogenesis. Immunofluorescence has visualized the colocalization of QBRICK and ECM3 on both apical and basal surface of ectoderm. On the basal surface, QBRICK and ECM3 constitute together a mesh-like fibrillar structure along the blastocoel wall. When the expression of ECM3 was knocked down by antisense-morpholino oligonucleotides, the ECM3-QBRICK fibrillar structure completely disappeared. When QBRICK was knocked down, the ECM3 was still present, but the basally localized fibers became fragmented. The ingression and migration of primary mesenchymal cells were not critically affected, but their migration at later stages was severely affected in both knock-down embryos. As a consequence of impaired primary mesenchymal cell migration, improper spicule formation was observed. These results indicate that ECM3 and QBRICK are components of extracellular matrix, which play important role in primary mesenchymal cell migration, and that sea urchin is a useful experimental animal model to investigate human disease-associated extracellular matrix proteins.
海胆胚胎一直是研究细胞行为的模式生物之一,因为它们的细胞组成简单且身体透明。它们还为我们提供了一个机会,研究在海胆中保守的人类感兴趣的蛋白质的分子功能。在这里,我们报告人类疾病相关的细胞外基质同源物 ECM3 和 QBRICK 对于海胆胚胎发生期间间质细胞迁移是必需的。免疫荧光已可视化 QBRICK 和 ECM3 在外胚层的顶端和基底表面上的共定位。在基底表面,QBRICK 和 ECM3 一起构成沿囊胚腔壁的网状纤维状结构。当 ECM3 的表达被反义 - 吗啉代寡核苷酸敲低时,ECM3-QBRICK 纤维状结构完全消失。当 QBRICK 被敲低时,ECM3 仍然存在,但基底定位的纤维变得碎片化。初级间质细胞的内陷和迁移没有受到严重影响,但在两种敲低胚胎中,它们在后期的迁移受到严重影响。由于初级间质细胞迁移受损,观察到不合适的骨针形成。这些结果表明,ECM3 和 QBRICK 是细胞外基质的组成部分,它们在初级间质细胞迁移中发挥重要作用,并且海胆是研究人类疾病相关细胞外基质蛋白的有用实验动物模型。