Name: a practical approach to genetic inducible fate mapping: a visual guide to mark and track cells in vivo
Uploaded: 2009-12-30T15:00:00
Description: Watch this Scientific Journal Video about A Practical Approach to Genetic Inducible Fate Mapping: A Visual Guide to Mark and Track Cells In Vivo at JoVE.com

S. mGFP小鼠阿伯尔和批判地阅读文件，并提供技术援助的Zervas实验室的成员，我们非常感谢。答：布朗支持大脑科学卡普兰夏季研究生研究奖。 E.诺曼德是支持的脑科学计划的研究生研究奖。这项研究是由启动研究经费（MZ）。

他莫昔芬的制备及灌胃程序（E8.5） <ul><li>通过溶解在他莫昔芬预热玉米油，准备了他莫昔芬20毫克/毫升的原液。 </li><li>解决方案在37℃，2小时一个nutator和涡间歇。 </li><li>他莫昔芬的股票免受光，储存在4 ° C，并使用长达1个月。 </li><li>命运映射实验，建立一个养殖对瑞士韦伯斯特女性（野生型;从Taconic公司购买）组成的和男性携带一个基因的特定CreER T等位基因和记者等位基因。...

<ol>
	<li>Ellisor, D., Koveal, D., Hagan, N., Brown, A., Zervas, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Comparative+analysis+of+conditional+reporter+alleles+in+the+developing+embryo+and+embryonic+nervous+system.">Comparative analysis of conditional reporter alleles in the developing embryo and embryonic nervous system.</a> Gene Expr Patterns. 9, 475-489 (2009).</li><li>Zervas, M., Millet, S., Ahn, S., Joyner, A. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cell+behaviors+and+genetic+lineages+of+the+mesencephalon+and+rhombomere+1.">Cell behaviors and genetic lineages of the mesencephalon and rhombomere 1.</a> Neuron. 43, 345-357 (2004).</li></ol>

GIFM系统，我们已经证明可以用于广泛的多种细胞过程中的问题的答案。例如，具有不同的Cre司机工程小鼠可以用来针对任何类型的细胞，组织或利益的遗传谱系。此外，因为他莫昔芬可在任何时间点胚胎或产后管理，重组，可以针对任何相关的发展阶段。该系统的空间和时间的控制是研究如何表达特定基因的细胞结构或地区的利益，以及细胞迁移和图案在开发过程中的事件作出贡献的理想。 ...

a practical approach to genetic inducible fate mapping: a visual guide to mark and track cells in vivo

命运的地图生成标记和跟踪体内细胞，以确定如何祖贡献的具体结构和细胞类型，在发展中国家和成人组织。在这个概念的发展是 G enetic我nducible F 吃中号apping（GIFM），将基因表达，细胞的命运，并在体内的细胞行为，创建基于遗传谱系的命运地图。 GIFM战功的的X CreER线，X是一个基因或基因调控元件的设置，赋予一个修改后的噬菌体蛋白质，Cre 重组酶（CreER T） 的表达空间 。CreER T包含一个修改的E strogen ř eceptor配体结合域呈现CreER T扣押在细胞质中的药物他莫昔芬的情况下。他莫昔芬发行CreER T，即转移到细胞核与调解由loxP位点两侧的DNA序列之间的重组结合。在GIFM，重组之间，通常会出现一个loxP位两侧停止前，如GFP报告基因的卡带。 小鼠繁殖包含一个区域或特定细胞类型的 CreER和有条件的记者等位基因。未经处理的小鼠不会有标记，因为在记者停止卡带防止进一步的报告基因的转录。我们管理灌胃定时怀孕的女性，这提供CreER T版本和随后易位到细胞核去除记者从停止磁带的时间控制的他莫昔芬。重组之后，记者等位基因的组成和heritably表达。这一系列事件标志着细胞等，他们的遗传史是不可磨灭的记录。重组记者，作为一个高保真的遗传谱系追踪器，一旦上，从最初用来驱动CreER T基因表达耦合。 我们申请GIFM在小鼠成年细胞类型和组织的遗传谱系研究的正常发展和确定贡献。我们也使用GIFM遵循细胞突变的遗传背景，以更好地了解复杂的表型，模仿人类遗传性疾病的显着特点。 此视频文章指南，通过实验方法的研究人员成功申请GIFM。我们证明的方法，使用特点Wnt1 CreER T;由 mGFP小鼠胚胎一天（五）8.5管理通过清扫在E12.5和epifluorescence立体显微镜分析灌胃他莫昔芬。我们还演示了如何微为外植体的制备或流式细胞仪分析解剖的命运映射域和成人命运映射的大脑解剖整装荧光成像。总的来说，这些程序允许研究人员在发育生物学和疾病模型解决的关键问题。

Watch this Scientific Journal Video about A Practical Approach to Genetic Inducible Fate Mapping: A Visual Guide to Mark and Track Cells In Vivo at JoVE.com

A Practical Approach to Genetic Inducible Fate Mapping: A Visual Guide to Mark and Track Cells In Vivo

遗传诱导的命运映射（GIFM）标记和细胞与精致的空间和时间的控制轨道<em&gt;在体内</em&gt;和阐发了如何从一个特定的遗传谱系细胞有助于发展中国家和成人组织。这里展示的是命运之图E12.5小鼠胚胎啶和外植体分析所需的技术。

一个实用的方法遗传诱导的命运映射：一个视觉引导标记和跟踪细胞在体内</em

Fate maps are generated by marking and tracking cells in vivo to determine how progenitors contribute to specific structures and cell types in developing ...

Research

JoVE Journal

Biology

A Practical Approach to Genetic Inducible Fate Mapping: A Visual Guide to Mark and Track Cells In Vivo

Method for Culture of Early Chick Embryos ex vivo (New Culture)

Method for Culture of Early Chick Embryos ex vivo New Culture

文化早期鸡胚体外（新文化）的方法

The chick embryo is a valuable tool in the study of early embryonic development. Its transparency, accessibility and ease of manipulation, make it an ...

科研

生物学

Double Whole Mount in situ Hybridization of Early Chick Embryos

双整装原位杂交技术在早期鸡胚

A Magnetic Tether System to Investigate Visual and Olfactory Mediated Flight Control in Drosophila

一个磁性的系绳系统，以调查在果蝇的视觉和嗅觉介导的飞行控制

It has been clear for many years that insects use visual cues to stabilize their heading in a wind stream. Many animals track odors carried in the wind. ...

A Reverse Genetic Approach to Test Functional Redundancy During Embryogenesis

一个反向遗传学方法测试胚胎发育过程中的功能冗余

Gene function during embryogenesis is typically defined by loss-of-function experiments, for example by targeted mutagenesis (knockout) in the mouse. In ...

Fate Mapping of Human Embryonic Stem Cells by Teratoma Formation

人类胚胎干细胞畸胎瘤形成命运的映射

Human embryonic stem cells (hESCs) have an unlimited capacity for self-renewal, and the ability to differentiate into cells derived from all three ...

Single Cell Fate Mapping in Zebrafish

单细胞在斑马鱼的命运映射

The ability to differentially label single cells has important implications in developmental biology. For instance, determining how hematopoietic, ...

Blastomere Explants to Test for Cell Fate Commitment During Embryonic Development

卵裂球外植体测试胚胎发育过程中细胞命运的承诺

Fate maps, constructed from lineage tracing all of the cells of an embryo, reveal  which tissues descend from each cell of the embryo.  Although fate maps ...

A Practical Guide to Phylogenetics for Nonexperts

实用指南系统发育的非专家

Many researchers, across incredibly diverse foci, are applying phylogenetics to their research question(s). However, many researchers are new to this ...

Validation of a Mouse Model to Disrupt LINC Complexes in a Cell-specific Manner

小鼠模型的验证扰乱LINC配合物在细胞特异性的方式

Nuclear migration and anchorage within developing and adult tissues relies heavily upon large macromolecular protein assemblies called LInkers of the ...

Genome-wide Mapping of Drug-DNA Interactions in Cells with COSMIC (Crosslinking of Small Molecules to Isolate Chromatin)

Genome-wide Mapping of Drug-DNA Interactions in Cells with COSMIC Crosslinking of Small Molecules to Isolate Chromatin

药物与DNA相互作用的细胞全基因组映射与COSMIC（小分子交联，染色质隔离）

The genome is the target of some of the most effective chemotherapeutics, but most of these drugs lack DNA sequence specificity, which leads to ...