Xenopus laevis Keller Explants.
Xenopus laevis Keller Explants.
复制标题
非洲爪蟾凯勒外植体。
DOI：
10.1101/pdb.prot4749
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发表时间：
2007
期刊：
影响因子：
--
通讯作者：
R. Harland
中科院分区：
文献类型：
--
作者：
H. Sive;R. Grainger;R. Harland
引言基本凯勒外植体是一个矩形的背中内胚层和外胚层早期原肠胚阶段非洲爪蟾胚胎。宽约60°至90°，从瓶状细胞延伸到动物极。本方案描述了如何解剖、组装和培养Keller外植体。凯勒外植体的目的最初是允许观察原肠胚形成运动，特别是收敛延伸，在文化。当外植体卷曲时，这是很难做到的，但在凯勒三明治中，外植体是平坦培养的，或者作为单片（开放面外植体），或者更常见的是作为两片夹在一起，它们的内表面并置（封闭三明治）。外植体在盖玻片碎片或置于硅真空油脂上的玻璃桥下培养，直到所需的阶段，通常在神经胚形成期间或之后。中胚层不是在外胚层下面内卷，而是与相邻的外胚层在一个平面上伸长。在原肠胚形成开始时，在外胚层和中胚层发生显著的垂直并置之前制作外植体。
INTRODUCTIONThe basic Keller explant is a rectangle of dorsal mesendoderm and ectoderm from an early-gastrula-stage Xenopus laevis embryo. It is ~60° to 90° wide, extending from the bottle cells to the animal pole. This protocol describes how to dissect, assemble, and cultivate Keller explants. The purpose of Keller explants was initially to allow observation of gastrulation movements, particularly convergent extension, in culture. This is difficult to do when explants curl up, but in Keller sandwiches, the explants are cultured flat, either as a single sheet (open-face explant) or more frequently as two sheets sandwiched together with their inner surfaces apposed (closed sandwich). Explants are cultured beneath a coverslip fragment or a glass bridge resting on silicone vacuum grease until the desired stage, usually during or after neurulation. Instead of involuting beneath the ectoderm, mesoderm elongates in a plane with adjacent ectoderm. Explants are made at the onset of gastrulation before significant vertical juxtaposition of ectoderm and mesoderm has occurred.