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乳胶珠作为软骨形成过程中细胞表面-细胞外基质相互作用的探针:纤连蛋白氨基末端肝素结合域作用的证据

Latex beads as probes of cell surface-extracellular matrix interactions during chondrogenesis: evidence for a role for amino-terminal heparin-binding domain of fibronectin.

作者信息

Frenz D A, Akiyama S K, Paulsen D F, Newman S A

机构信息

Department of Cell Biology and Anatomy, New York Medical College, Valhalla 10595.

出版信息

Dev Biol. 1989 Nov;136(1):87-96. doi: 10.1016/0012-1606(89)90132-2.

Abstract

Fibronectin-rich mesenchymal condensations form at sites of incipient chondrogenesis in the developing vertebrate limb, and in cultures of limb bud mesenchyme. We have used 6 microns polystyrene latex beads coated with various substances as probes for adhesive interactions that may mediate the formation of these condensations. Beads coated with heparin, chondroitin sulfate, or poly L-lysine, that were mixed with limb bud mesenchymal cells were centripetally conveyed into fibronectin-rich regions of cell condensation over a period of several days. Beads coated with dextran sulfate remained uniformly dispersed throughout the cultures during the same period. A monoclonal antibody directed against the amino-terminal heparin-binding domain of fibronectin completely inhibited accumulation of heparin-coated beads at condensing foci, but monoclonal antibodies directed against the collagen- or cell-binding domains of fibronectin were not inhibitory. Accumulation of chondroitin sulfate- or poly L-lysine-coated beads at condensing foci was unaffected by the antibody against the fibronectin amino terminus. Peptides with the sequence arg-gly-asp-ser or gly-arg-gly-asp-ser, which inhibit adhesive interactions mediated by the integrin-binding domain of fibronectin, had no effect on conveyance or accumulation of heparin-coated beads, but the peptide with the sequence gly-arg-gly, a repeated motif in the amino-terminal heparin-binding domain was completely inhibitory. These findings indicate that the amino-terminal heparin-binding domain of fibronectin can, within a tissue microenvironment, interact adhesively with heparin-like components on the surfaces of polystyrene beads, and by implication, on mesenchymal cells themselves. This interaction may therefore be a component of the condensation-forming mechanism in chondrogenic mesenchyme.

摘要

富含纤连蛋白的间充质凝聚物在发育中的脊椎动物肢体的初始软骨形成部位以及肢体芽间充质培养物中形成。我们使用涂有各种物质的6微米聚苯乙烯乳胶珠作为可能介导这些凝聚物形成的粘附相互作用的探针。与肢体芽间充质细胞混合的涂有肝素、硫酸软骨素或聚L-赖氨酸的珠子在几天内被向心地输送到富含纤连蛋白的细胞凝聚区域。在同一时期,涂有硫酸葡聚糖的珠子在整个培养物中保持均匀分散。一种针对纤连蛋白氨基末端肝素结合域的单克隆抗体完全抑制了肝素包被珠子在凝聚灶的积累,但针对纤连蛋白胶原或细胞结合域的单克隆抗体没有抑制作用。硫酸软骨素或聚L-赖氨酸包被珠子在凝聚灶的积累不受抗纤连蛋白氨基末端抗体的影响。具有精氨酸-甘氨酸-天冬氨酸-丝氨酸或甘氨酸-精氨酸-甘氨酸-天冬氨酸-丝氨酸序列的肽,可抑制由纤连蛋白整合素结合域介导的粘附相互作用,对肝素包被珠子的输送或积累没有影响,但具有甘氨酸-精氨酸-甘氨酸序列的肽,即氨基末端肝素结合域中的重复基序,具有完全抑制作用。这些发现表明,纤连蛋白的氨基末端肝素结合域在组织微环境中可以与聚苯乙烯珠表面的肝素样成分发生粘附相互作用,由此推断,也可以与间充质细胞本身发生粘附相互作用。因此,这种相互作用可能是软骨形成间充质中凝聚物形成机制的一个组成部分。

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