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乳胶珠作为神经嵴通路的探针:层粘连蛋白、胶原蛋白和表面电荷对珠子移位的影响。

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

作者信息

Bronner-Fraser M

出版信息

J Cell Biol. 1984 Jun;98(6):1947-60. doi: 10.1083/jcb.98.6.1947.

Abstract

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.

摘要

在鸟类胚胎的躯干区域,神经嵴细胞沿两条路径迁移:一条是在背侧,紧贴外胚层下方;另一条是在腹侧,位于神经管和体节之间。本实验室先前的研究表明,将未包被的乳胶珠注射到幼胚后,它们能够沿着腹侧神经嵴路径移位;然而,包被纤连蛋白的珠子则被限制在腹侧路径之外(Bronner - Fraser, M.E., 1982, 《发育生物学》, 91: 50 - 63)。在此,我们拓展这些观察结果,以确定其他大分子对珠子分布的影响。数据显示,包被层粘连蛋白的珠子,与包被纤连蛋白的珠子一样,被限制在腹侧路径之外。相反,包被I型胶原的珠子在注射后会向腹侧移位。由于大分子具有特定的电荷特性,包被珠子所导致的表面电荷变化可能会混淆对结果的解释。为了可预测地改变初始表面电荷,通过用多聚氨基酸包被乳胶珠来研究静电对珠子移动的影响。测量了包被氨基酸聚合物或各种具有生物学重要性的大分子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移位能力。包被多聚赖氨酸的珠子(带正电荷)与包被纤连蛋白和层粘连蛋白的珠子一样,被限制在腹侧路径之外,尽管纤连蛋白和层粘连蛋白珠子都是带负电荷的。相反,包被聚酪氨酸的珠子(带中性电荷)与带负电荷的包被胶原或未包被的珠子一样向腹侧移位。结果表明,乳胶珠表面的电荷特性与其随后沿腹侧路径移位的能力之间不存在相关性。因此,一种独立于表面电荷效应的黏附机制必定可以解释乳胶珠在神经嵴路径上的受限或移位情况。

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