Gevers Petra, Coenen Anthony J M, Schipper Henk, Stroband Henri W J, Timmermans Lucy P M
Department of Animal Morphology and Cell Biology, Agricultural University, Marijkeweg 40, P.O. Box 338, 6709 PG, Wageningen, The Netherlands.
Department of Animal Physiology, University of Nijmegen, Toernooiveld, 6525, ED, Nijmegen, The Netherlands.
Rouxs Arch Dev Biol. 1993 Feb;202(3):152-158. doi: 10.1007/BF00365305.
The present report firstly describes a pilot study in which, during early development of embryos of the common carp, Cyprinus carpio, the cellular adhesion to fibronectin (FN) was blocked by administration of GRGDS peptide (which binds to the FN-receptor). As this treatment resulted in developmental aberrations, suggesting a functional role for FN, the major part of the work was focussed on the distribution of reactivity of anti-FN antibodies during epiboly and gastrulation. GRGDS treatment had a concentration dependent effect on development. Incubation of embryos in 1.5 mg/ml from the 32-cell stage onwards caused a retardation of epiboly, which did not proceed beyond 60%. The embryos did not show involution, as was confirmed by histological study. These preliminary results suggest that FN is involved in both epiboly and gastrulation of carp embryos. During cleavage, no specific extracellular binding of anti-FN antiserum could be observed. However, binding to a number of cell membranes took place from early epiboly onwards. After the onset of gastrulation, we observed a gradually increasing number of the deepest epiblast cells, showing immunostaining on part of their surface, facing the yolk syncytial layer (YSL) or the involuted cells. During early epiboly, anti-FN binding was restricted to areas in front of the migratory hypoblast cells. Later on, binding was found at the border of hypoblast and epiblast cells. At 100% epiboly, some contact areas of epiblast and hypoblast showed a discontinuous lining of reactivity, whilst other areas appeared devoid of anti-FN binding sites. The results indicate that FN is involved in the migration and guidance of hypoblast cells during gastrulation in carp.
本报告首先描述了一项初步研究，在鲤鱼（Cyprinus carpio）胚胎的早期发育过程中，通过给予GRGDS肽（其与纤连蛋白受体结合）来阻断细胞与纤连蛋白（FN）的粘附。由于这种处理导致发育异常，表明FN具有功能性作用，因此大部分工作集中在抗FN抗体在卵裂和原肠胚形成过程中的反应性分布上。GRGDS处理对发育具有浓度依赖性影响。从32细胞期开始，将胚胎置于1.5 mg/ml的溶液中孵育会导致卵裂延迟，卵裂进程不会超过60%。组织学研究证实，胚胎未出现内卷。这些初步结果表明，FN参与了鲤鱼胚胎的卵裂和原肠胚形成过程。在卵裂期间，未观察到抗FN抗血清的特异性细胞外结合。然而，从早期卵裂开始，就有一些细胞膜发生了结合。原肠胚形成开始后，我们观察到最深层的上胚层细胞数量逐渐增加，其部分表面朝向卵黄合胞体层（YSL）或内卷细胞的表面呈现免疫染色。在早期卵裂期间，抗FN结合仅限于迁移的下胚层细胞前方的区域。后来，在下胚层和上胚层细胞的边界处发现了结合。在卵裂100%时，上胚层和下胚层的一些接触区域显示出反应性的不连续排列，而其他区域似乎没有抗FN结合位点。结果表明，FN参与了鲤鱼原肠胚形成过程中下胚层细胞的迁移和引导。
