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）。这个时间随不同的地段的胶原酶。因此，一小时后，一个小样本的卵母细胞，在解剖显微镜下研究。正常DE - folliculated卵母细胞应彼此分开，应该很容易将其插入到卵母细胞玻璃针。如果不充分去folliculated的卵母细胞，他们留在胶原酶溶液，并检查了一遍，每五分钟。 </li><li>洗净的卵母细胞与修改巴特的生理盐水（按揭证券化：88 mM的氯化钠，1 mM的氯化钾0.82 mM的硫酸镁4，0.33 mM的钙（NO 3）2，0.41 mM的氯化钙2，10毫米HEPES，pH值7.5）的三倍，和转让以一个100毫米直径的培养皿中含有按揭证券，另加1％青霉素/链霉素。现在已经准备好被显微注射的卵母细胞。如果将在另一天进行显微注射​​，储存长达一个星期的卵母细胞在4 ° C。 </li><li>使用解剖显微镜，选择成熟的阶段，六，卵母细胞显微注射。这些卵母细胞是大的，黑色的动物半球和南半球的奶油色的植物之间良好的对比度。 </li><li>六，卵母细胞的成熟阶段转移到一个微孔板（NUNC，以及体积10μL）。这项工作应仔细，使用吸管尖端已被切成末，允许不间断的卵母细胞吸200微升移液器。 </li></ol>第2部分： 爪蟾卵母细胞显微注射<ol><li>准备进口基材，加入少量的1％溴酚蓝，援助在显微注射的可视化显微注射。例如，我们添加1μL溴酚蓝10μL底物。 </li><li>广场上一个直径为100毫米的培养皿封口膜的小片，并免除的解决方案上的封口膜带注入5μL下降。 </li><li>被注入的解决方案，填写以前校准的注射针头的注入+ MATIC拖轮拉6.6μL微量德拉蒙德获得。对于这一点，微量愿望模式。要校准注射针头，针点标记每0.5毫米，对应的50 NL量。 </li><li>细胞质注射针尖插入一个卵母细胞在植物性半球，动物半球非常接近，在一个约45度角。转动的微量设置microinject，进口基材NL 50 microinject每个卵母细胞。针点的标记是用来监测量已注入基板。 </li><li>注入卵母细胞转移到一个小的（直径35毫米）的培养皿填充与按揭证券，并在室温孵化所需的时间量（时间点的选择，以便观察与NPC的相关进口基材，发生这种情况10至30分钟，正在积极向全国人民代表大会运输蛋白质和病毒，这一次也要看）注射部位和大小的蛋白质/病毒。 </li><li>当孵化完成后，转让的卵母细胞，以4毫升的玻璃小瓶，其中包含MBS 2％戊二醛，并固定在4℃过夜° C。 </li></ol>第3部分：夹层爪蟾卵母细胞<ol><li>第二天，洗卵母细胞与MBS的3倍。 </li><li>卵母细胞转移到一个小的培养皿中充满低盐缓冲（LSB：1毫米氯化钾， 氯化镁 2 0.5毫米，10毫米的HEPES，pH值7.5） 。使用解剖显微镜和解剖镊子，请从每个卵母细胞的植物极。它是有用的稳定解剖一个镊子的卵母细胞，而另一对是用于去除的植物极像剪刀。在这一点上是可能的，以评估在细胞质中的蓝色色调的存在显微注射成功。该协议是继续只对卵母细胞显微注射成功。此外，如果样本准备为少数族裔，这清扫步骤使得它更容易找到修整和切片的样品时，原子核。 </li><li>再次修复解剖的卵母细胞，在室温下1小时，用2％戊二醛的LSB。 </li><li>固定后，洗解剖的卵母细胞与LSB三次。 </li></ol>第4部分：嵌入和薄切片的EM注入卵母细胞的制备<ol><li>解剖卵母细胞转移到抑郁症的幻灯片。吸为液体作为POSsible，然后迅速（让他们不要干出来的）覆盖2％低熔点琼脂糖解剖的卵母细胞。虽然琼脂糖仍是软的，用一个吸头彼此分开的卵母细胞，以确保每一个卵母细胞是面临向上或向下（侧身）。 </li><li>允许琼脂糖巩固约10分钟。琼脂糖凝固后，用刀片切成小块，每个都包含一个解剖卵母细胞的琼脂糖。 </li><li>放置在一个4毫升的玻璃小瓶琼脂糖包埋的卵母细胞，修复后一小时在室温下用1％的官方机密条例“ 中的LSB 4。 </li><li>洗净样品在LSB的3倍。如果有必要，样品保存在4 ° C过夜，第二天继续协议。 </li><li>依序脱水（每次20分钟50，70，和90％的乙醇）的样品中的酒精升序系列。这是由两个100％的乙醇的变化，每次15分钟。 100％丙酮15分钟，最后脱水样本。 </li><li> 1:1混合的EPON（Fluka公司）和丙酮为一小时2:1的EPON和丙酮的混合了两个小时，至少6个小时，终于在纯EPON通过渗透，渗透样品。 </li><li>充满了新鲜的纯EPON放置在平坦嵌入模具样品。卵母细胞在这样一种方式，面向细胞核注射部位接近方 - 方已解剖的卵母细胞 - 在这种情况下，将切片第一。这一步优化，可视化的选择基板的进口机会。 </li><li>最后，EPON的聚合两天在60 ° C </li><li>为了形象化的样品，块修剪和使用ultramicrotome然后切片。部分转移到电磁电网，染色使用的标准程序，以及可视化与透射电子显微镜。，我们将不会显示这个协议的一部分，因为它是标准的 EM程序。 </li></ol>第5部分：代表性的成果： 如果该协议已获得成功，那么核信封（NE）和NPC应的EM显微照片清晰可见。根据在基板上注射，注射和固定之间的时间量，基板应在全国人民代表大会的细胞质面临中可见，全国人民代表大会，或在核面对全国人民代表大会。 图1显示了与（三）相邻的细胞质和细胞核（N）已注入杆状病毒AcMNPV的衣壳从爪蟾卵母细胞培养在4 ° C四小时，东北横截面，并嵌入和薄片处理新兴市场作为描述。箭头指向的NPC。在全国人大细胞质面对衣壳对接是一个白色箭头指示。 相比之下，图2显示了一个东北横截面，并与邻近的细胞质（C）和细胞核从已注射的小鼠细小分钟病毒（MVM）四个小时，在室温下孵育的爪蟾卵母细胞（ N）处理嵌入和新兴市场作为描述的薄片。使用这种技术，我们发现，MVM的诱导NE（Cohen和盘特，2005年）的中断。括号表示在NE的休息。箭头指向的NPC。假定与NE的MVM衣壳是由白色箭头指示。 <img src="/files/ftp_upload/1106/1106_fig1.png" alt="图1" width="600" height="459" /> 图1：一个NE与相邻的细胞质（C）和细胞核（N）从爪蟾卵母细胞已注射杆状病毒AC MNPV衣壳，孵育4截面的电子显微镜° C为四个小时，并处理嵌入和薄片EM所述。箭头指向的NPC。衣壳是由白色箭头指示。比例尺：100纳米。 <img src="/files/ftp_upload/1106/figure2.png" alt="图2" width="600" height="264" /> 图2：一个NE与相邻的细胞质（C）和细胞核从已注射的小鼠细小分钟病毒（MVM）四个小时，在室温下孵育的爪蟾卵母细胞（N）的横截面的电子显微镜，处理嵌入和新兴市场作为描述的薄片。箭头指向的NPC。括号表示在NE的休息。假定与NE的MVM衣壳是由白色箭头指示。比例尺：100纳米。

<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

22.9K 次观看.  University of British Columbia - UBC. 在这里，我们展示了胞浆内显微注射的非洲爪蟾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。

视频: Microinjection of Xenopus Laevis Oocytes

Plastic Embedding and Sectioning of Xenopus laevis Embryos

Plastic sections maintain true tissue morphology in thin sections of tissue that can be immunostained with fluorescent secondary antibodies, making this method more useful than paraffin-embedded or frozen sections for many types of tissue. The method for staining, plastic embedding, and sectioning is demonstrated in this video.

塑料嵌入和非洲爪蟾胚胎切片

科研

生物学

Obtaining Eggs from Xenopus laevis Females

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, collect and dejelly the eggs, and sort the eggs to remove any damaged eggs.

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, collect and dejelly the eggs, and sort the eggs to remove any damaged eggs.

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

Crude and fractionated Xenopus egg extracts can be used to provide ingredients for reconstituting cellular processes for morphological and biochemical analysis. Egg lysis and differential centrifugation are used to prepare the crude extract which in turn in used to prepare fractionated extracts and light membrane preparations.

Crude and fractionated Xenopus egg extracts can be used to provide ingredients for reconstituting cellular processes for morphological and biochemical analysis. Egg lysis and differential centrifugation are used to prepare the crude extract which in turn in used to prepare fractionated extracts and light membrane preparations.

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

Since its development by Sakmann and Neher 1, 2, the patch clamp has become established as an extremely useful technique for electrophysiological measurement of single or multiple ion channels in cells. This technique can be applied to ion channels in both their native environment and expressed in heterologous cells, such as oocytes harvested from the African clawed frog, Xenopus laevis. Here, we describe the well-established technique of patch clamp recording from Xenopus oocytes. This technique is used to measure the properties of expressed ion channels either in populations (macropatch) or individually (single-channel recording). We focus on techniques to maximize the quality of oocyte preparation and seal generation. With all factors optimized, this technique gives a probability of successful seal generation over 90 percent. The process may be optimized differently by every researcher based on the factors he or she finds most important, and we present the approach that have lead to the greatest success in our hands.

This is intended as an introduction to patch clamp recording from Xenopus laevis oocytes. It covers vitelline membrane removal, formation of a gigaohm seal (gigaseal), and the optional conversion of the patch to the outside-out topology.

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

RNA localization is a conserved mechanism of establishing cell polarity. Vg1 mRNA localizes to the vegetal pole of Xenopus laevis oocytes and acts to spatially restrict gene expression of Vg1 protein. Tight control of Vg1 distribution in this manner is required for proper germ layer specification in the developing embryo. RNA sequence elements in the 3' UTR of the mRNA, the Vg1 localization element (VLE) are required and sufficient to direct transport. To study the recognition and transport of Vg1 mRNA in vivo, we have developed an imaging technique that allows extensive analysis of trans-factor directed transport mechanisms via a simple visual readout. 
To visualize RNA localization, we synthesize fluorescently labeled VLE RNA and microinject this transcript into individual oocytes. After oocyte culture to allow transport of the injected RNA, oocytes are fixed and dehydrated prior to imaging by confocal microscopy. Visualization of mRNA localization patterns provides a readout for monitoring the complete pathway of RNA transport and for identifying roles in directing RNA transport for cis-acting elements within the transcript and trans-acting factors that bind to the VLE (Lewis et al., 2008, Messitt et al., 2008). We have extended this technique through co-localization with additional RNAs and proteins (Gagnon and Mowry, 2009, Messitt et al., 2008), and in combination with disruption of motor proteins and the cytoskeleton (Messitt et al., 2008) to probe mechanisms underlying mRNA localization.

Visualizing RNA Localization in Xenopus Oocytes

Visualization of in vivo RNA transport is accomplished by microinjection of fluorescently labeled RNA transcripts into Xenopus oocytes, followed by confocal microscopy.

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

Studying the contribution of maternally inherited molecules to vertebrate early development is often hampered by the time and expense necessary to generate maternal-effect mutant animals. Additionally, many of the techniques to overexpress or inhibit gene function in organisms such as Xenopus and zebrafish fail to sufficiently target critical maternal signaling pathways, such as Wnt signaling. In Xenopus, manipulating gene function in cultured oocytes and subsequently fertilizing them can ameliorate these problems to some extent. Oocytes are manually defolliculated from donor ovary tissue, injected or treated in culture as desired, and then stimulated with progesterone to induce maturation. Next, the oocytes are introduced into the body cavity of an ovulating host female frog, whereupon they will be translocated through the host's oviduct and acquire modifications and jelly coats necessary for fertilization. The resulting embryos can then be raised to the desired stage and analyzed for the effects of any experimental perturbations. This host-transfer method has been highly effective in uncovering basic mechanisms of early development and allows a wide range of experimental possibilities not available in any other vertebrate model organism.

Fertilization of Xenopus oocytes using the Host Transfer Method

Procedure for fertilizing Xenopus oocytes by 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

Many proteins play a dual role in embryonic development. Those that regulate cell fate determination in a specific tissue can also affect the development of a larger region of the embryo. This makes defining its role in a particular tissue difficult to analyze. For example, noggin overexpression in Xenopus laevis embryos causes the expansion of the entire anterior region, including the eye1,2. From this result, it is not known if Noggin plays a direct role in eye determination or that by causing an expansion of neural tissue, Noggin indirectly affects eye formation. Having this complex phenotype makes studying its eye-specific role in cell fate determination difficult to analyze. We have developed an assay that overcomes this problem. Taking advantage of the pluripotent nature of the Xenopus laevis animal cap 3, we have developed an assay to test the ability of gene product(s), like noggin or the eye field transcription factors (EFTFs), to transform caps into particular tissue or cell types by transplanting this tissue onto the side of the embryo 4. While we have found either Noggin protein treatment or a collection of transcription factors can determine retinal cell fate in animal caps, this procedure could be used to identify gene product(s) involved in specifying other tissues as well.

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

Animal caps overexpressing gene product(s) are transplanted to the flank of developing Xenopus laevis embryos in order to establish whether tissue is determined.

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

Embryonic epithelial cells serve as an ideal model to study morphogenesis where multi-cellular tissues undergo changes in their geometry, such as changes in cell surface area and cell height, and where cells undergo mitosis and migrate. Furthermore, epithelial cells can also regulate morphogenetic movements in adjacent tissues1. A traditional method to study epithelial cells and tissues involve chemical fixation and histological methods to determine cell morphology or localization of particular proteins of interest. These approaches continue to be useful and provide "snapshots" of cell shapes and tissue architecture, however, much remains to be understood about how cells acquire specific shapes, how various proteins move or localize to specific positions, and what paths cells follow toward their final differentiated fate. High resolution live imaging complements traditional methods and also allows more direct investigation into the dynamic cellular processes involved in the formation, maintenance, and morphogenesis of multicellular epithelial sheets. Here we demonstrate experimental methods from the isolation of animal cap tissues from Xenopus laevis embryos to confocal imaging of epithelial cells and simple measurement approaches that together can augment molecular and cellular studies of epithelial morphogenesis.

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

Xenopus embryonic epithelia are an ideal model system to study cell behaviors such as polarity development and shape change during epithelial morphogenesis. Traditional histology of fixed samples is increasingly being complemented by live-cell confocal imaging. Here we demonstrate methods to isolate frog tissues and visualize live epithelial cells and their cytoskeleton using live-cell confocal microscopy.

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

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 stages of embryonic development, and to define how enhancers and promoters regulate gene expression within the embryo. The protocol demonstrated here can be used to efficiently produce transgenic Xenopus laevis embryos. This transgenesis approach involves three parts: 1. Sperm nuclei are isolated from adult X. laevis testis by treatment with lysolecithin, which permeabilizes the sperm plasma membrane. 2. Egg extract is prepared by low speed centrifugation, addition of calcium to cause the extract to progress to interphase of the cell cycle, and a high-speed centrifugation to isolate interphase cytosol. 3. Nuclear transplantation: the nuclei and extract are combined with the linearized plasmid DNA to be introduced as the transgene and a small amount of restriction enzyme. During a short reaction, egg extract partially decondenses the sperm chromatin and the restriction enzyme generates chromosomal breaks that promote recombination of the transgene into the genome. The treated sperm nuclei are then transplanted into unfertilized eggs. Integration of the transgene usually occurs prior to the first embryonic cleavage such that the resulting embryos are not chimeric. These embryos can be analyzed without any need to breed to the next generation, allowing for efficient and rapid generation of transgenic embryos for analyses of promoter and gene function. Adult X. laevis resulting from this procedure also propagate the transgene through the germline and can be used to generate lines of transgenic animals for multiple purposes.

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

This video protocol demonstrates a method for generating transgenic Xenopus laevis by introduction of transgenes into sperm nuclei followed by nuclear transplantation into unfertilized eggs.

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

Craniofacial birth defects occur in 1 out of every 700 live births, but etiology is rarely known due to limited understanding of craniofacial development. To identify where signaling pathways and tissues act during patterning of the developing face, a 'face transplant' technique has been developed in embryos of the frog Xenopus laevis. A region of presumptive facial tissue (the "Extreme Anterior Domain" (EAD)) is removed from a donor embryo at tailbud stage, and transplanted to a host embryo of the same stage, from which the equivalent region has been removed. This can be used to generate a chimeric face where the host or donor tissue has a loss or gain of function in a gene, and/or includes a lineage label. After healing, the outcome of development is monitored, and indicates roles of the signaling pathway within the donor or surrounding host tissues. Xenopus is a valuable model for face development, as the facial region is large and readily accessible for micromanipulation. Many embryos can be assayed, over a short time period since development occurs rapidly. Findings in the frog are relevant to human development, since craniofacial processes appear conserved between Xenopus and mammals.

Facial Transplants in Xenopus laevis Embryos

A technique for transplanting "Extreme Anterior Domain" facial tissue between Xenopus laevis embryos has been developed. Tissue can be moved from one gene expression background into another, allowing the study of local requirements for craniofacial development and for signaling interactions between facial regions.

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. ...