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Latex beads as probes of a neural crest pathway: effects of laminin, collagen, and surface charge on bead translocation

机译:乳胶珠作为神经c通路的探针:层粘连蛋白,胶原蛋白和表面电荷对珠转运的影响

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摘要

In the trunk region of avian embryos, neural crest cells migrate along two pathways: dorsally just under the ectoderm, and ventrally between the neural tube and the somites. Previous work from this laboratory has shown that uncoated latex beads are able to translocate along the ventral neural crest pathway after injection into young embryos; however, beads coated with fibronectin are restricted from the ventral route ( Bronner -Fraser, M.E., 1982, Dev. Biol., 91: 50-63). Here, we extend these observations to determine the effects of other macromolecules on bead distribution. The data show that laminin-coated beads, like fibronectin-coated beads, are restricted from the ventral pathway. In contrast, beads coated with type I collagen translocate ventrally after injection. Because macromolecules have characteristic charge properties, changes in surface charge caused by coating the beads may confound interpretation of the results. Electrostatic effects on bead movement were examined by coating the latex beads with polyamino acids in order to predictably alter the initial surface charge. The surface charge before injection was measured for beads coated with amino acid polymers or with various biologically important macromolecules; the subsequent translocation ability of these beads was then monitored in the embryo. Polylysine-coated beads (positively charged) were restricted from the ventral pathway as were fibronectin and laminin-coated beads, even though fibronectin and laminin beads were both negatively charged. In contrast, polytyrosine -coated beads ( neutrally charged) translocated ventrally as did negatively charged collagen-coated or uncoated beads. The results demonstrate that no correlation exists between the charge properties on the latex bead surface and their subsequent ability to translocate along the ventral pathway. Therefore, an adhesion mechanism independent of surface charge effects must explain the restriction or translocation of latex beads on a neural crest pathway.
机译:在禽类胚胎的躯干区域,神经cells细胞沿着两条途径迁移:背侧正好位于外胚层下方,而腹侧则位于神经管和体节之间。该实验室先前的工作表明,未包被的乳胶珠在注入年轻胚胎后能够沿着腹侧神经c通路移位。然而,用纤连蛋白包被的珠子从腹侧途径受到限制(Bronner-Fraser,M.E.,1982,Dev.Biol。,91:50-63)。在这里,我们扩展这些观察以确定其他大分子对珠分布的影响。数据表明,层粘连蛋白包被的小珠,像纤连蛋白包被的小珠一样,受到腹侧通路的限制。相反,注射后,涂有I型胶原蛋白的珠子在腹腔内移位。因为大分子具有特征性的电荷特性,所以由包覆珠粒引起的表面电荷变化可能会混淆结果的解释。通过用聚氨基酸包被乳胶珠来检查对珠运动的静电影响,以便可预测地改变初始表面电荷。测量了用氨基酸聚合物或各种生物学上重要的大分子包被的珠子在注射前的表面电荷;然后在胚胎中监测这些珠子随后的易位能力。尽管纤连蛋白和层粘连蛋白珠均带负电,但聚赖氨酸包被的珠(带正电荷)与纤连蛋白和层粘连蛋白的珠一样受到腹侧通路的限制。相反,聚酪氨酸涂层的珠子(带中性电荷)与带负电荷的胶原蛋白涂层或未涂层的珠子一样在腹侧移位。结果表明,乳胶珠表面上的电荷性质与其随后沿腹侧途径移位的能力之间不存在相关性。因此,独立于表面电荷效应的粘附机制必须解释乳胶珠在神经c通路上的限制或易位。

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    Bronner-Fraser Marianne;

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  • 年度 1984
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