Johnson K E, Darribère T, Boucaut J C
Department of Anatomy, George Washington University Medical Center, Washington, D.C. 20037.
Dev Biol. 1990 Jan;137(1):86-99. doi: 10.1016/0012-1606(90)90010-g.
Rana pipiens eggs fertilized by Rana esculenta sperm (ESC) hybrid embryos develop until gastrulation in control Rana pipiens embryos (PIP) and then show morphogenetic arrest. After arrest, ESC do not gastrulate but live for 5 days as blastula-like embryos. We studied the distribution of fibronectin (FN)-containing fibrils and integrin (INT) in PIP and ESC. There are many FN-fibrils in PIP organized in anastomosing networks radiating away from the center of individual cells and across intercellular boundaries. ESC have fewer fibrils compared to PIP. These fibrils are first located between cells in disorganized arrays. After arrest in ESC, when PIP are Stage 14 neurulae, many more FN-fibrils appear. INT-staining occurs in both embryos in similar patterns. In xenoplastic transplantations, we found that the extracellular matrix on the inner surface of the ESC blastocoel roof serves as a substratum for PIP cell migration. In an in vitro assay, we found more cell adhesion to FN-substrata in PIP than in ESC. Cell locomotion rates on FN-substrata were 1.70 +/- 0.85 microns/min for PIP but only 0.46 +/- 0.56 microns/min for ESC. We also found that the inner surface of the blastocoel roof from ESC can not promote cell adhesion and locomotion when Stage 11 fragments are used for conditioning but that Stage 14 fragments can deposit a FN-fibril-rich extracellular matrix which supports PIP mesodermal cell migration at a rate of 1.26 +/- 0.38 microns/min.
用食用蛙精子(ESC)使豹蛙卵受精后,杂交胚胎在对照豹蛙胚胎(PIP)中发育至原肠胚形成阶段,然后出现形态发生停滞。停滞之后,ESC不会形成原肠胚,而是以囊胚样胚胎的形式存活5天。我们研究了PIP和ESC中含纤连蛋白(FN)的纤维和整合素(INT)的分布情况。PIP中有许多FN纤维,它们以从单个细胞中心辐射并穿过细胞间边界的吻合网络形式组织起来。与PIP相比,ESC中的纤维较少。这些纤维最初以无序排列位于细胞之间。在ESC出现停滞时,当PIP处于14期神经胚时,会出现更多的FN纤维。INT染色在两个胚胎中的模式相似。在异种移植中,我们发现ESC囊胚腔顶内表面的细胞外基质可作为PIP细胞迁移的底物。在体外试验中,我们发现PIP对FN底物的细胞黏附比ESC更多。PIP在FN底物上的细胞运动速率为1.70±0.85微米/分钟,而ESC仅为0.46±0.56微米/分钟。我们还发现,当使用11期片段进行预处理时,ESC囊胚腔顶的内表面不能促进细胞黏附和运动,但14期片段可以沉积富含FN纤维的细胞外基质,该基质支持PIP中胚层细胞以1.26±0.38微米/分钟的速率迁移。