利用TUNEL检测蜘蛛胚胎细胞死亡。

CSH protocols Pub Date : 2008-10-01 DOI:10.1101/pdb.prot5069
Nikola-Michael Prpic, Michael Schoppmeier, Wim G M Damen
{"title":"利用TUNEL检测蜘蛛胚胎细胞死亡。","authors":"Nikola-Michael Prpic,&nbsp;Michael Schoppmeier,&nbsp;Wim G M Damen","doi":"10.1101/pdb.prot5069","DOIUrl":null,"url":null,"abstract":"<p><p>INTRODUCTIONThe spider Cupiennius salei, commonly known as the American Wandering Spider, is a particularly useful laboratory model 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 in itself will be a necessary cornerstone for comparing the different metazoan phyla, including chordates. A feature of apoptosis (i.e., cell death) is the cleavage or fragmentation of DNA that occurs in dead or dying cells. This protocol describes the detection of fragmented DNA in whole-mount Cupiennius embryos. The 3'-OH ends of these DNA fragments can be labeled with the terminal deoxynucleotidyl-transferase-mediated dUTP-digoxigenin nick-end labeling (TUNEL) technique. This protocol uses a terminal deoxynucleotidyl transferase to add labeled dUTP to the fragmented DNA, and this label is then detected by immunocytochemistry. The TUNEL technique is a relatively easy way to obtain a reliable picture of the cell death pattern during normal and abnormal development.</p>","PeriodicalId":10835,"journal":{"name":"CSH protocols","volume":null,"pages":null},"PeriodicalIF":0.0000,"publicationDate":"2008-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1101/pdb.prot5069","citationCount":"3","resultStr":"{\"title\":\"Detection of Cell Death in Spider Embryos Using TUNEL.\",\"authors\":\"Nikola-Michael Prpic,&nbsp;Michael Schoppmeier,&nbsp;Wim G M Damen\",\"doi\":\"10.1101/pdb.prot5069\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><p>INTRODUCTIONThe spider Cupiennius salei, commonly known as the American Wandering Spider, is a particularly useful laboratory model 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 in itself will be a necessary cornerstone for comparing the different metazoan phyla, including chordates. A feature of apoptosis (i.e., cell death) is the cleavage or fragmentation of DNA that occurs in dead or dying cells. This protocol describes the detection of fragmented DNA in whole-mount Cupiennius embryos. The 3'-OH ends of these DNA fragments can be labeled with the terminal deoxynucleotidyl-transferase-mediated dUTP-digoxigenin nick-end labeling (TUNEL) technique. This protocol uses a terminal deoxynucleotidyl transferase to add labeled dUTP to the fragmented DNA, and this label is then detected by immunocytochemistry. The TUNEL technique is a relatively easy way to obtain a reliable picture of the cell death pattern during normal and abnormal development.</p>\",\"PeriodicalId\":10835,\"journal\":{\"name\":\"CSH protocols\",\"volume\":null,\"pages\":null},\"PeriodicalIF\":0.0000,\"publicationDate\":\"2008-10-01\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://sci-hub-pdf.com/10.1101/pdb.prot5069\",\"citationCount\":\"3\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"CSH protocols\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"https://doi.org/10.1101/pdb.prot5069\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"\",\"JCRName\":\"\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"CSH protocols","FirstCategoryId":"1085","ListUrlMain":"https://doi.org/10.1101/pdb.prot5069","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
引用次数: 3

摘要

蜘蛛Cupiennius salei,通常被称为美国流浪蜘蛛,是胚胎学研究中特别有用的实验室模型,因为研究和操纵其胚胎发育的工具是可用的。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。这本身将是比较不同后生动物门(包括脊索动物)的必要基础。细胞凋亡(即细胞死亡)的一个特征是发生在死亡或垂死细胞中的DNA分裂或断裂。本方案描述了在全胚库比尼乌斯胚胎中DNA片段的检测。这些DNA片段的3'-OH末端可以用末端脱氧核苷酸转移酶介导的dutp -地高igenin镍端标记(TUNEL)技术进行标记。该方案使用末端脱氧核苷酸转移酶将标记的dUTP添加到片段DNA上,然后通过免疫细胞化学检测该标记。TUNEL技术是一种相对简单的方法,可以获得正常和异常发育过程中细胞死亡模式的可靠图像。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Detection of Cell Death in Spider Embryos Using TUNEL.

INTRODUCTIONThe spider Cupiennius salei, commonly known as the American Wandering Spider, is a particularly useful laboratory model 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 in itself will be a necessary cornerstone for comparing the different metazoan phyla, including chordates. A feature of apoptosis (i.e., cell death) is the cleavage or fragmentation of DNA that occurs in dead or dying cells. This protocol describes the detection of fragmented DNA in whole-mount Cupiennius embryos. The 3'-OH ends of these DNA fragments can be labeled with the terminal deoxynucleotidyl-transferase-mediated dUTP-digoxigenin nick-end labeling (TUNEL) technique. This protocol uses a terminal deoxynucleotidyl transferase to add labeled dUTP to the fragmented DNA, and this label is then detected by immunocytochemistry. The TUNEL technique is a relatively easy way to obtain a reliable picture of the cell death pattern during normal and abnormal development.

求助全文
通过发布文献求助,成功后即可免费获取论文全文。 去求助
来源期刊
自引率
0.00%
发文量
0
×
引用
GB/T 7714-2015
复制
MLA
复制
APA
复制
导出至
BibTeX EndNote RefMan NoteFirst NoteExpress
×
提示
您的信息不完整,为了账户安全,请先补充。
现在去补充
×
提示
您因"违规操作"
具体请查看互助需知
我知道了
×
提示
确定
请完成安全验证×
copy
已复制链接
快去分享给好友吧!
我知道了
右上角分享
点击右上角分享
0
联系我们:info@booksci.cn Book学术提供免费学术资源搜索服务,方便国内外学者检索中英文文献。致力于提供最便捷和优质的服务体验。 Copyright © 2023 布克学术 All rights reserved.
京ICP备2023020795号-1
ghs 京公网安备 11010802042870号
Book学术文献互助
Book学术文献互助群
群 号:481959085
Book学术官方微信