Detection of Cell Proliferation in Spider Embryos Using BrdU Labeling.

CSH protocols Pub Date : 2008-10-01 DOI:10.1101/pdb.prot5071
Nikola-Michael Prpic, Michael Schoppmeier, Wim G M Damen
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引用次数: 2

Abstract

INTRODUCTIONThe spider Cupiennius salei, commonly known as the American wandering spider, is particularly useful for embryological studies because of the availability of tools to study and manipulate its embryonic development. Cupiennius is used to study axis formation, segmentation, appendage development, neurogenesis, and silk production. These studies contribute to our understanding of the evolution of these processes, but they also help us to understand the origin and diversification of evolutionary novelties. Comparisons between spiders and insects can show the degree of conservation and divergence of developmental mechanisms during arthropod evolution. Any embryological feature conserved between spiders and insects is likely to represent an ancestral feature for arthropods. Comparative molecular embryological work in insects and spiders should eventually allow us to define a molecular archetype for the phylum Arthropoda. This will be a necessary cornerstone for comparing the different metazoan phyla, including chordates. This protocol describes the detection of proliferating cells in whole-mount Cupiennius embryos. When labeled nucleotides are introduced into mitotically dividing cells, these cells incorporate the labels into the newly synthesized DNA. Thus, only cells that have synthesized DNA after the addition of the label will be detected. This protocol uses 5-bromo-2'-deoxy-uridine (BrdU) as a label that is subsequently detected by immunocytochemistry. BrdU labeling is a relatively easy way to detect cells that have recently synthesized DNA. The main advantage of this technique is that the label accumulates over time and, by varying the incubation time before fixation, an increasingly cumulative picture of cell proliferation activity can be obtained.

利用BrdU标记检测蜘蛛胚胎细胞增殖。
蜘蛛Cupiennius salei,通常被称为美国流浪蜘蛛,对于胚胎学研究特别有用,因为可以使用工具来研究和操纵其胚胎发育。Cupiennius被用来研究轴的形成、分割、附属物的发育、神经发生和丝的产生。这些研究有助于我们理解这些过程的进化,但它们也有助于我们理解进化新事物的起源和多样化。蜘蛛和昆虫之间的比较可以显示节肢动物进化过程中发育机制的保守程度和分化程度。蜘蛛和昆虫之间的任何胚胎学特征都可能代表节肢动物的祖先特征。昆虫和蜘蛛的分子胚胎学比较工作最终将使我们能够确定节肢动物门的分子原型。这将是比较包括脊索动物在内的不同后生动物门的必要基础。本方案描述了在整座秋葵胚胎中增殖细胞的检测。当标记的核苷酸被引入有丝分裂的细胞时,这些细胞将标记结合到新合成的DNA中。因此,只有在添加标签后合成DNA的细胞才会被检测到。该方案使用5-溴-2'-脱氧尿苷(BrdU)作为标记,随后通过免疫细胞化学检测。BrdU标记是检测最近合成DNA的细胞的一种相对简单的方法。该技术的主要优点是,标记随着时间的推移而积累,并且通过改变固定前的孵育时间,可以获得细胞增殖活性的逐渐累积的图像。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
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