Name: microinjection of xenopus laevis oocytes
Uploaded: 2009-02-23T00:00:00
Description: Watch this Scientific Journal Video about Microinjection of Xenopus Laevis Oocytes at JoVE.com

我们感谢大卫Theilmann（太平洋农业食品研究中心，Summerland，不列颠哥伦比亚省）提供杆状病毒AcMNPV的和有益的讨论。 这项工作是支持由加拿大创新基金会（CFI），加拿大卫生研究所（CIHR）健康研究（MSFHR），迈克尔史密斯基金会的机构，自然科学与工程研究理事会（NSERC），加拿大的补助金。

第1部分：爪蟾卵母细胞显微注射的制备<ol><li>放置到一个50毫升锥形中的钙，免费修改巴特的生理盐水20毫升胶原酶的解决方案（5 mg / ml的胶原酶的试管一个爪蟾卵巢小片（约2厘米）：88 mM的氯化钠，1 mM的氯化钾，0.82 mM的硫酸镁 ，10毫米HEPES，pH值7.5）。胶原酶用于去除环绕的卵母细胞的卵泡细胞。 </li><li>摇床平台上放置管，轻轻地摇动一小时（在100 RPM）。这个时间随不同...

<ol>
	<li>Pant&#233;, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Use+of+intact+Xenopus+oocytes+in+nucleocytoplasmic+transport+studies.">Use of intact Xenopus oocytes in nucleocytoplasmic transport studies.</a> Methods Mol. Biol. 322, 301-314 (2006).</li><li>Pant&#233;, N., Kann, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Nuclear+pore+complex+is+able+to+transport+macromolecules+with+diameters+of+about+39+nm.">Nuclear pore complex is able to transport macromolecules with diameters of about 39 nm.</a> Mol. Biol. Cell. 13 (2), 425-434 (2002).</li><li>Rabe, B., Vlachou, A., Pant&#233;, N., Helenius, N., Kann, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Nuclear+import+of+hepatitis+B+virus+capsids+and+release+of+the+viral+genome.">Nuclear import of hepatitis B virus capsids and release of the viral genome.</a> Proc. Natl. Acad. Sci. U.S.A. 100 (17), 9849-9854 (2003).</li><li>Cohen, S., Pant&#233;, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Pushing+the+Envelope:+Microinjection+of+MVM+into+Xenopus+oocytes+causes+damage+to+the+nuclear+envelope.">Pushing the Envelope: Microinjection of MVM into Xenopus oocytes causes damage to the nuclear envelope.</a> J. Gen. Virol. 86 (12), 3243-3252 (2005).</li><li>Pant&#233;, N., Jarmolowski, A., Izaurralde, E., Sauder, U., Baschong, W., Mattaj, I. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Visualizing+nuclear+export+of+different+classes+of+RNA+by+electron+microscopy.">Visualizing nuclear export of different classes of RNA by electron microscopy.</a> RNA. 3, 498-513 (1997).</li></ol>

薄切片新兴市场相结合的爪蟾卵母细胞显微注射是一个非常有效的工具，为研究核质运输。已使用该系统，地图，通过全国人民代表大会不同的步骤，例如与细胞质的细丝和核篮（盘特，2006年审查），如全国人民代表大会的结构组成部分的核进口基板相互作用，进口。它也被用来研究核复制病毒的核进口（盘特和卡恩，2002年， 拉贝等人，2003 年 ，科恩和盘特，2005年） 。 ...

microinjection of xenopus laevis oocytes

显微注射爪蟾卵母细胞超薄切片电子显微镜（EM）是一个优秀的系统研究核质运输。由于其庞大的核和核孔复合物（筹备）的高密度，核运输中可以很容易地在爪蟾卵母细胞的可视化。通过这个系统的使用（2006年盘特，审查）已获得很大的启示成核的进口和出口的机制。此外，我们用爪蟾卵母细胞显微注射剖析几种病毒的核进口途径，在宿主细胞核中复制。 这里我们展示了一个拥有核武器的进口基板的爪蟾卵母细胞胞浆内显微注射。我们还准备注入卵母细胞的可视化显示薄切片新兴市场，包括清扫，脱水和卵母细胞中嵌入环氧树脂嵌入树脂。最后，我们提供已显微注射的苜蓿californica nucleopolyhedrovirus杆状病毒（AcMNPV的）或小鼠（MVM）的细小分钟病毒的衣壳，并讨论该技术的潜在应用卵母细胞的代表性成果。

Watch this Scientific Journal Video about Microinjection of Xenopus Laevis Oocytes at JoVE.com

Microinjection of Xenopus Laevis Oocytes

在这里，我们展示了胞浆内显微注射的<em&gt;非洲爪蟾</em&gt;卵母细胞的核进口基材，以及准备注入卵母细胞超薄切片电子显微镜可视化。

非洲爪蟾卵母细胞的微量注射

Microinjection of Xenopus laevis oocytes followed by thin-sectioning electron microscopy (EM) is an excellent system for studying nucleocytoplasmic ...

Research

JoVE Journal

Biology

Plastic Embedding and Sectioning of Xenopus laevis Embryos

塑料嵌入和非洲爪蟾胚胎切片

科研

生物学

Obtaining Eggs from Xenopus laevis Females

从非洲爪蟾女性获得卵子

The eggs of Xenopus laevis intact, lysed, and/or fractionated are useful for a wide variety of experiments. This protocol shows how to induce egg laying, ...

Preparation and Fractionation of Xenopus laevis Egg Extracts

制备及分馏爪蟾卵提取

Crude and fractionated Xenopus egg extracts can be used to provide ingredients for reconstituting cellular processes for morphological and biochemical ...

Patch Clamp Recording of Ion Channels Expressed in  Xenopus Oocytes

补丁钳爪蟾卵母细胞中表达的离子通道

Since its development by Sakmann and Neher 1, 2, the patch clamp has become established as an extremely useful technique for electrophysiological ...

Visualizing RNA Localization in Xenopus Oocytes

Visualizing RNA Localization in Xenopus Oocytes

可视化的RNA中的本地化爪蟾卵母细胞

RNA localization is a conserved mechanism of establishing cell polarity.  Vg1 mRNA localizes to the vegetal pole of Xenopus laevis oocytes and acts to ...

Fertilization of Xenopus oocytes using the Host Transfer Method

Fertilization of Xenopus oocytes using the Host Transfer Method

施肥爪蟾卵母细胞使用主机传输方法

Studying the contribution of maternally inherited molecules to vertebrate early development is often hampered by the time and expense necessary to ...

Tissue Determination Using the Animal Cap Transplant (ACT) Assay in Xenopus laevis

Tissue Determination Using the Animal Cap Transplant ACT Assay in Xenopus laevis

组织测定动物第移植（ACT）的含量非洲爪蟾</em

Many proteins play a dual role in embryonic development. Those that regulate cell fate determination in a specific tissue can also affect the development ...

Live-cell Imaging and Quantitative Analysis of Embryonic Epithelial Cells in Xenopus laevis

Live-cell Imaging and Quantitative Analysis of Embryonic Epithelial Cells in Xenopus laevis

活细胞成像和胚胎上皮细胞的定量分析非洲爪蟾</em

Embryonic epithelial cells serve as an ideal model to study morphogenesis where multi-cellular tissues undergo changes in their geometry, such as changes ...

Production of Transgenic Xenopus laevis by Restriction Enzyme Mediated Integration and Nuclear Transplantation

Production of Transgenic Xenopus laevis by Restriction Enzyme Mediated Integration and Nuclear Transplantation

生产转基因非洲爪蟾限制性内切酶介导的整合和核移植

Stable integration of cloned gene products into the Xenopus genome is necessary to control the time and place of expression, to express genes at later ...

Facial Transplants in Xenopus laevis Embryos

Facial Transplants in Xenopus laevis Embryos

在面部移植<em&gt;非洲爪蟾</em&gt;胚胎

Craniofacial birth defects occur in 1 out of every 700 live births, but etiology is rarely known due to limited understanding of craniofacial development. ...