Name: lens transplantation in zebrafish and its application in the analysis of eye mutants
Uploaded: 2009-06-01T00:00:00
Description: Watch this Scientific Journal Video about Lens Transplantation in Zebrafish and its Application in the Analysis of Eye Mutants at JoVE.com

此过程如下很大程度上条例草案“杰弗里实验室移植技术开发洞穴鱼。

第1部分：准备胚胎在这个协议中，我们将使用JJ XY符号来表示一个假设的斑马鱼镜头突变。 <ol><li>斑马鱼品系均保持在28.5在14H light/10h暗周期彗星的标准鱼设施条件。 </li><li>在晚上，斑马鱼应变JJ XY地方的男性和女性：AB / TU; TG（mGFP）在坦克与一个分频器，它们彼此分开。这种交叉的后代将表达绿色荧光蛋白基因，在所有组织和捐助者将?...

<ol>
	<li>Thut, C. J., Rountree, R. B., Hwa, M., Kingsley, D. 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+large-scale+in+situ+screen+provides+molecular+evidence+for+the+induction+of+eye+anterior+segment+structures+by+the+developing+lens.">A large-scale in situ screen provides molecular evidence for the induction of eye anterior segment structures by the developing lens.</a> Dev Biol. 231, 63-76 (2001).</li><li>Yamamoto, Y., Jeffery, W. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Central+role+for+the+lens+in+cave+fish+eye+degeneration.">Central role for the lens in cave fish eye degeneration.</a> Science. 289, 631-633 (2000).</li><li>Easter, S. S., Nicola, G. N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+development+of+vision+in+the+zebrafish(Danio+rerio).">The development of vision in the zebrafish(Danio rerio).</a> Dev Biol. 180, 646-663 (1996).</li><li>Schmitt, E., Dowling, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Early+eye+morphogenesis+in+the+zebrafish.+Brachydanio+rerio.">Early eye morphogenesis in the zebrafish. Brachydanio rerio.</a> J.comp. Neuro. 344 (4), 532-542 (1994).</li><li>Malicki, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Harnessing+the+power+of+forward+genetics--analysis+of+neuronal+diversity+and+patterning+in+the+zebrafish+retina.">Harnessing the power of forward genetics--analysis of neuronal diversity and patterning in the zebrafish retina.</a> Trends Neurosci. 23 (11), 531-541 (2000).</li><li>Malicki, J., Pujic, Z., Thisse, C., Thisse, B., Wei, X. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Forward+and+reverse+genetic+approaches+to+the+analysis+of+eye+development+in+zebrafish.">Forward and reverse genetic approaches to the analysis of eye development in zebrafish.</a> Vision Res. 42 (4), 527-533 (2002).</li><li>Avanesov, A., Malicki, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Approaches+to+study+neurogenesis+in+the+zebrafish+retina.">Approaches to study neurogenesis in the zebrafish retina.</a> Methods Cell Biol. 76, 333-384 (2004).</li><li>Vihtelic, T. S., Hyde, D. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Zebrafish+mutagenesis+yields+eye+morphological+mutants+with+retinal+and+lens+defects.">Zebrafish mutagenesis yields eye morphological mutants with retinal and lens defects.</a> Vision Res. 42, 535-540 (2002).</li><li>Godinho, L., Mumm, J. S., Williams, P. R., Schroeter, E. H., Koerber, A., Park, S. W., Leach, S. D., Wong, R. O. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Targeting+of+amacrine+cell+neurites+to+appropriate+synaptic+laminae+in+the+developing+zebrafish+retina.">Targeting of amacrine cell neurites to appropriate synaptic laminae in the developing zebrafish retina.</a> Development. Nov 132, 5069-5079 (2005).</li></ol>

有几个步骤需要特别注意在镜头移植。 <ol><li>锐利的针：这是更好地开展晶状体移植前的准备几针，针的尖端应尽可能薄。一个较厚的针将撕裂更因为它的直径更广泛的组织，造成眼睛未能愈合。 </li><li>安排的胚胎，移植前，您必须胚胎的位置，所以他们就躺在他们身边。当定位，1.2％琼脂糖硬化速度非常快。要缓慢琼脂糖硬化，培​​养皿可以让它漂浮在40℃水浴预热。 </li><li>插入主机眼?...

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<td>0.1mm diameter tungsten wire</td>
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lens transplantation in zebrafish and its application in the analysis of eye mutants

镜头视杯[1,2]的发展起着重要的作用。使用斑马鱼作为模式生物，关于镜头发展的问题可以得到解决。斑马鱼是由于一些优势特点，包括体积小，繁殖力高，生命周期短，便于照顾的遗传研究非常有用。镜头发展迅速出现在斑马鱼。 72 HPF，斑马鱼的镜头功能成熟[3 ]。丰富的遗传和分子资源来支持在斑马鱼的研究。此外，斑马鱼眼的其他脊椎动物相似，提供其作为人类缺陷极好的动物模型 [4-7]使用的基础。一些斑马鱼突变体表现出镜头的异常，包括高水平的细胞死亡，在某些情况下会导致晶状体组织的一个完整的变性[8 ]。 为了确定是否镜头异常是由于内在的原因或与周围组织有缺陷的相互作用，一个突变成野生型眼晶状体移植。仔细解剖用火抛光的金属针，突变型或野生型镜头是从供体动物，并转移到主机。为了区分野生型和突变型的组织，转基因线是用来作为供体。该行表示在所有组织的膜结合型绿色荧光蛋白，包括镜头。这种移植技术在斑马鱼的镜头突变体的研究是必不可少的工具。

Watch this Scientific Journal Video about Lens Transplantation in Zebrafish and its Application in the Analysis of Eye Mutants at JoVE.com

Lens Transplantation in Zebrafish and its Application in the Analysis of Eye Mutants

镜头的发展涉及到与其他组织的互动。几个的斑马鱼眼突变体的特点是异常的小镜头尺寸。在这里，我们展示了一个镜头移植实验，以确定这是否表型是由于内在的原因，或环绕镜头的组织，有缺陷的相互作用。

在斑马鱼和镜头移植在眼突变分析中的应用

The lens plays an important role in the development of the optic cup[1,2]. Using the zebrafish as a model organism, questions regarding lens development ...

Research

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在成年斑马鱼的全肾骨髓移植

Hematopoietic stem cells (HSC) are a rare population of pluripotent cells that maintain all the differentiated blood lineages throughout the life of an ...

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The Ladder Rung Walking Task: A Scoring System and its Practical Application.

梯级步行任务：一个评分系统，其实际应用。

Progress in the development of animal models for/stroke, spinal cord injury, and other neurodegenerative disease requires tests of high sensitivity to ...

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The anterior surface of the ocular lens is covered by a monolayer of epithelial cells, which proliferate in an annular zone underlying the ciliary body.  ...

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原代培养及质粒电穿孔尔蒂小鼠器官。

In all mammals, the sensory epithelium for audition is located along the spiraling organ of Corti that resides within the conch shaped cochlea of the ...

In vivo Electroporation of Morpholinos into the Adult Zebrafish Retina

In vivo Electroporation of Morpholinos into the Adult Zebrafish Retina

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The zebrafish has become an important in vivo model in biomedical research. Effective methods must be developed and utilized to deliver compounds or ...

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The circadian clock regulates many aspects of life, including sleep, locomotor activity, and body temperature (BTR) rhythms1,2. We recently identified a ...

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The zebrafish model has emerged as a relevant system to study kidney development, regeneration and disease. Both the embryonic and adult zebrafish kidneys ...

Sequential Application of Glass Coverslips to Assess the Compressive Stiffness of the Mouse Lens: Strain and Morphometric Analyses

盖玻片序贯应用评估鼠标透镜的抗压刚度：应变和形态分析

The eye lens is a transparent organ that refracts and focuses light to form a clear image on the retina. In humans, ciliary muscles contract to deform the ...

High-resolution Time-lapse Imaging and Automated Analysis of Microtubule Dynamics in Living Human Umbilical Vein Endothelial Cells

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The physiological process by which new vasculature forms from existing vasculature requires specific signaling events that trigger morphological changes ...