Name: live imaging of apoptotic cell clearance during drosophila embryogenesis
Uploaded: 2013-08-18T14:00:00
Description: Watch this Scientific Journal Video about Live Imaging of Apoptotic Cell Clearance during Drosophila Embryogenesis at JoVE.com

支持这项工作是由居里夫人重返格兰特（IRG249084）。感谢所有成员的Kurant实验室。

要按照两群细胞凋亡细胞清除体内必须标明：吞噬细胞和细胞凋亡的动态。 为了纪念我们使用了含有果蝇线的的模拟cytGFP标记，专门吞噬细胞在胚胎标签：巨噬细胞，神经胶质细胞和外胚层8吞噬细胞群。人们可以使用不同的标记为吞噬细胞，其中含有一个血球特定的Gal4的驱动程序（CRQ-GAL4）的行或神经胶质细胞特异性Gal4的驱动（ 回购Ga...

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	<li>Kinchen, J. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+model+to+die+for:+signaling+to+apoptotic+cell+removal+in+worm,+fly+and+mouse.">A model to die for: signaling to apoptotic cell removal in worm, fly and mouse.</a> Apoptosis. 15, 998-1006 (2010).</li><li>Kinchen, J. M., Ravichandran, K. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Phagosome+maturation:+going+through+the+acid+test.">Phagosome maturation: going through the acid test.</a> Nat. Rev. Mol. Cell Biol. 9, 781-795 (2008).</li><li>Kinchen, J. M., Ravichandran, K. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Phagocytic+signaling:+you+can+touch,+but+you+can't+eat.">Phagocytic signaling: you can touch, but you can't eat.</a> Curr. Biol. 18, 521-524 (2008).</li><li>Stuart, L. M., Ezekowitz, R. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Phagocytosis:+elegant+complexity.">Phagocytosis: elegant complexity.</a> Immunity. 22, 539-550 (2005).</li><li>Lauber, K., Blumenthal, S. G., Waibel, M., Wesselborg, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Clearance+of+apoptotic+cells:+getting+rid+of+the+corpses.">Clearance of apoptotic cells: getting rid of the corpses.</a> Mol. Cell. 14, 277-287 (2004).</li><li>Henson, P. M., Hume, D. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Apoptotic+cell+removal+in+development+and+tissue+homeostasis.">Apoptotic cell removal in development and tissue homeostasis.</a> Trends Immunol. 27, 244-250 (2006).</li><li>Kurant, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Keeping+the+CNS+clear:+glial+phagocytic+functions+in+Drosophila.">Keeping the CNS clear: glial phagocytic functions in Drosophila.</a> Glia. 59, 1304-1311 (2011).</li><li>Kurant, E., Axelrod, S., Leaman, D., Gaul, U. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Six-microns-under+acts+upstream+of+Draper+in+the+glial+phagocytosis+of+apoptotic+neurons.">Six-microns-under acts upstream of Draper in the glial phagocytosis of apoptotic neurons.</a> Cell. 133, 498-509 (2008).</li><li>Mergliano, J., Minden, J. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Caspase-independent+cell+engulfment+mirrors+cell+death+pattern+in+Drosophila+embryos.">Caspase-independent cell engulfment mirrors cell death pattern in Drosophila embryos.</a> Development. 130, 5779-5789 (2003).</li><li>van den Eijnde, S. M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cell+surface+exposure+of+phosphatidylserine+during+apoptosis+is+phylogenetically+conserved.">Cell surface exposure of phosphatidylserine during apoptosis is phylogenetically conserved.</a> Apoptosis. 3, 9-16 (1998).</li></ol>

凋亡细胞的间隙是一个关键的细胞凋亡，这是高度动态的最后阶段。因此，实时研究的过程中是非常重要的。在这里，我们描述了一个协议，使凋亡细胞清除在监视居住开发果蝇胚胎。在此过程中，必须使用特定标记的凋亡细胞和吞噬细胞的标记两个细胞群体。该模拟cytGFP记者是适合于监测吞噬细胞的巨噬细胞，神经胶质细胞和外胚层，同时在相同的胚胎， 瓦特 HICH使得它可?...

适当的消除不需要的或异常的细胞通过细胞凋亡及随后的吞噬作用，胚胎发育，以及在成年组织的平衡是至关重要的。细胞吞噬凋亡细胞或凋亡细胞的清除是一个高度动态的过程，这四个步骤进行：（1） 招聘吞噬细胞的细胞凋亡（&#39;发现&#39;），（2） 承认的细胞吞噬的目标（ &#39;吃&#39;）和（3）的吞噬 ，随后由（4） 吞噬体的成熟和退化的细胞凋亡粒子1-5。吞噬细胞有两种类型：“专业的”巨噬细胞和未成熟树突状细胞，和“非专业组织驻地相邻的细胞，这是至关重要的凋亡细胞的清除过程中后生动物发展6,7。 在果蝇发育，消除多余的细胞通过细胞凋亡发生在3米的主要阶段，首先在中期到晚期胚胎，然后在蛹中旬，并再次在成年早期。胚胎发育过程中细胞凋亡的颗粒被去除由专业的吞噬细胞，巨噬细胞，由“非专业外胚层和神经胶质8,9。 在我们以前的工作中，我们已经确定了，六微米下吞噬受体（捷联惯组），这是吞噬细胞在胚胎发育过程中表达。我们使用模拟子产生吞噬细胞的人群（ 模拟cytGFP的 ）使我们能够监视巨噬细胞，外胚层和神经胶质细胞，同时在现场发育中的胚胎8的特异性指标。此外，通过使用在不同阶段的细胞凋亡中细胞凋亡的特异性标志物，我们能够遵循的凋亡细胞的清除在体内的动态。

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 <td>OncoImmunin</td>
 <td>A304R2G-5</td>
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 <td>LysoTracker</td>
 <td>Molecular Probes</td>
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 <td>93100</td>
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live imaging of apoptotic cell clearance during drosophila embryogenesis

适当的消除不必要的或异常细胞通过细胞凋亡和随后吞噬（凋亡细胞间隙）在所有后生生物的正常发育是至关重要的。凋亡细胞的清除是一个高度动态的过程密切相关的细胞死亡;勉强看到unengulfed凋亡细胞在体内正常情况下。为了了解凋亡细胞清除的不同步骤和比较“专业”的巨噬细胞-巨噬细胞和树突状细胞的&#39;非专业&#39; -组织居民相邻的细胞在体内的实时成像的过程，是极其宝贵的。在这里，我们描述了一个协议，在现场的果蝇胚胎研究凋亡细胞清除。要遵循不同的步骤，在我们使用特定的标记凋亡细胞和巨噬细胞的吞噬功能的动态。此外，我们可以监视两个巨噬细胞系统并行：“专业”的巨噬细胞和半专业图片ssional在发展中的中枢神经系统（CNS）的神经胶质细胞。这里介绍的方法采用了果蝇胚胎作为一个很好的模型实时研究凋亡细胞清除。

从动画中的表示帧，其中细胞凋亡被标记的膜联蛋白V，和神经胶质细胞，外胚层和巨噬细胞标记的模拟cytGFP示于图2。每个帧是3片各2μm的投影。 正确定位显示在中间的胚胎中枢神经系统标记的神经胶质细胞（G）胚胎阶段15。巨噬细胞（M），其中大多是以外的中枢神经系统，表现出强劲的细胞质GFP表达。外胚层细胞也标记细胞质GFP（E）。许多Annexin V的阳性...

Watch this Scientific Journal Video about Live Imaging of Apoptotic Cell Clearance during Drosophila Embryogenesis at JoVE.com

Live Imaging of Apoptotic Cell Clearance during Drosophila Embryogenesis

在这里，我们描述了一个有效的方法，研究凋亡细胞间隙的动态<em&gt;在体内</em&gt;。这种方法采用生活<em&gt;果蝇</em&gt;作为一个强大的监控模型，使用特定的标记凋亡细胞和巨噬细胞吞噬的凋亡细胞胚胎。

凋亡细胞清除过程中实时成像<em&gt;果蝇</em&gt;胚胎发育

The  proper elimination of unwanted or aberrant cells through apoptosis and  subsequent phagocytosis (apoptotic cell clearance) is crucial for normal  ...

Research

JoVE Journal

Biology

Live Imaging of Apoptotic Cell Clearance during Drosophila Embryogenesis

Live Imaging of Glial Cell Migration in the Drosophila Eye Imaginal Disc

住在果蝇眼睛成虫光盘胶质细胞迁移成像

Glial cells of both vertebrate and invertebrate organisms must migrate to final target regions in order to ensheath and support associated neurons.  While ...

科研

生物学

Live Imaging Of Drosophila melanogaster Embryonic Hemocyte Migrations

Live Imaging Of Drosophila melanogaster Embryonic Hemocyte Migrations

实时成像果蝇胚胎血细胞迁移

Many studies address cell migration using in vitro methods, whereas the physiologically relevant environment is that of the organism itself. Here we ...

Preparation of Developing and Adult Drosophila Brains and Retinae for Live Imaging

Preparation of Developing and Adult Drosophila Brains and Retinae for Live Imaging

发展和成人的制备果蝇大脑和视网膜实时成像

The Drosophila brain and visual system are widely utilized model systems to study neuronal development, function and degeneration. Here we show three ...

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 ...

Preparing Individual Drosophila Egg Chambers for Live Imaging

Preparing Individual Drosophila Egg Chambers for Live Imaging

准备个人果蝇蛋钱伯斯实时成像

Live cell imaging is an important technique applied to a number of Drosophila tissues used as models to investigate topics such as axis specification, ...

Live Imaging of GFP-labeled Proteins in Drosophila Oocytes

Live Imaging of GFP-labeled Proteins in Drosophila Oocytes

GFP标记的蛋白质在实时成像<em&gt;果蝇</em&gt;卵母细胞

The  Drosophila oocyte has been established as a versatile system for investigating  fundamental questions such as cytoskeletal function, cell ...

4D Imaging of Protein Aggregation in Live Cells

在活细胞中蛋白质聚集的4D成像

One of the key tasks of any living cell is maintaining the proper folding of newly synthesized proteins in the face of ever-changing environmental ...

The Tomato/GFP-FLP/FRT Method for Live Imaging of Mosaic Adult Drosophila Photoreceptor Cells

The Tomato/GFP-FLP/FRT Method for Live Imaging of Mosaic Adult Drosophila Photoreceptor Cells

番茄/ GFP-FLP / FRT方法马赛克成人的实时成像<em&gt;果蝇</em&gt;感光细胞

The Drosophila eye is widely used as a model  for studies of development and neuronal degeneration. With the powerful mitotic  recombination technique, ...

Cytological Analysis of Spermatogenesis: Live and Fixed Preparations of Drosophila Testes

Cytological Analysis of Spermatogenesis: Live and Fixed Preparations of Drosophila Testes

精子发生的细胞学分析：Live和固定制剂<em&gt;果蝇</em&gt;睾丸

Drosophila melanogaster is a powerful model system that has been widely used to elucidate a variety of biological processes. For example, studies of both ...

In Vivo Biosensor Tracks Non-apoptotic Caspase Activity in Drosophila

In Vivo Biosensor Tracks Non-apoptotic Caspase Activity in Drosophila

体内生物传感器跟踪非凋亡胱天蛋白酶的活动在果蝇

Caspases are the key mediators of apoptotic cell death via their proteolytic activity. When caspases are activated in cells to levels detectable by ...