Name: making gynogenetic diploid zebrafish by early pressure
Uploaded: 2009-06-30T00:00:00
Description: Watch this Scientific Journal Video about Making Gynogenetic Diploid Zebrafish by Early Pressure at JoVE.com

Charline沃克在乔治Streisinger的实验室在俄勒冈大学在20世纪70年代和80年代开发的斑马鱼，在体外受精和雌核发育二倍体的方法。这里所描述的方法是从Charline的协议，该协议是在斑马鱼图书 3不变。 Charline沃克在乔治Streisinger的实验室在俄勒冈大学在20世纪70年代和80年代开发的斑马鱼，在体外受精和雌核发育二倍体的方法。这里所描述的方法是从Charline的协议，该协议是在斑马鱼图书 3不变。

初压概述注：很多，如tricaine，鱼水和生理盐水汉克斯在此协议中所使用的试剂配方是在第10章在网上提供的斑马鱼的<a href="http://zfin.org/zf_info/zfbook/zfbk.html">书</a> 3 （http://zfin.org/zf_info/zfbook/zfbk.html） <ol><li>生产用紫外线灭活精子体外受精的胚胎。 </li><li>充满蛋水的边缘玻璃小瓶（塑料管理单元第）立即将受精卵。盖与乳胶薄板小瓶打开，并捕捉到小瓶在橡胶的顶部?...

<ol>
	<li>Streisinger, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Segregation+analyses+and+gene-centromere+distances+in+zebrafish.">Segregation analyses and gene-centromere distances in zebrafish.</a> Genetics. 112 (2), 311-311 (1986).</li><li>Streisinger, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Production+of+clones+of+homozygous+diploid+zebra+fish+(Brachydanio+rerio).">Production of clones of homozygous diploid zebra fish (Brachydanio rerio).</a> Nature. 291 (5813), 293-293 (1981).</li><li>Westerfield, 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> The Zebrafish Book: A guide for the laboratory use of zebrafish (Danio rerio). , (2000).</li><li>Johnson, S. L., Africa, D., Horne, S., Postlethwait, J. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Half-tetrad+analysis+in+zebrafish:+mapping+the+ros+mutation+and+the+centromere+of+linkage+group+I.">Half-tetrad analysis in zebrafish: mapping the ros mutation and the centromere of linkage group I.</a> Genetics. 139 (4), 1727-1727 (1995).</li><li>Beattie, C. E., Raible, D. W., Henion, P. D., Eisen, 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=Early+pressure+screens.">Early pressure screens.</a> Methods Cell Biol. 237 (71), 2575-25 (1999).</li><li>Johnson, S. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Centromere-linkage+analysis+and+consolidation+of+the+zebrafish+genetic+map.">Centromere-linkage analysis and consolidation of the zebrafish genetic map.</a> Genetics. 142 (4), 1277-1277 (1996).</li><li>Trede, N. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Zebrafish+mutants+with+disrupted+early+T-cell+and+thymus+development+identified+in+early+pressure+screen.">Zebrafish mutants with disrupted early T-cell and thymus development identified in early pressure screen.</a> Dev Dyn. 237 (9), 2575-2575 (1999).</li></ol>

重要的是要确保用这种方法产生的胚胎，是真正的雌核发育二倍体。作为对照，产生了正常的二倍体卵子（保持预留一些无紫外线灭活精子）和单倍体胚胎（保持不通过EP程序预留的紫外线精子受精卵离合器）离合器的离合器。在1天受精后单倍体胚胎体轴短，不规则的脑和体节的形态。单倍体是不是可行，经过2-3天。 EP二倍体应该看起来像正常的二倍体，虽然一些不规则的“EP怪物”可能会发生?...

<!DOCTYPE html PUBLIC "-//W3C//DTD XHTML 1.0 Transitional//EN" "http://www.w3.org/TR/xhtml1/DTD/xhtml1-transitional.dtd">
<html xmlns="http://www.w3.org/1999/xhtml" xml:lang="en" lang="en">	
		
		<div>
		<table border="1">
		<thead>
		<tr>
		<th>Material Name</th>
		<th>Type</th>
		<th>Company</th>
		<th>Catalogue Number</th>
		<th>Comment</th>
		</tr>
		</thead>
		<tbody>
		
<tr>
<td>Tricaine</td>
<td>&nbsp;</td>
<td>Sigma</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>Watch glass</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>French Press</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>Pressure cylinder</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>Instant ocean</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>Pasteur pipettes</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>Cut off the narrow end, fire polish</td>
</tr>
<tr>
<td>UV cross-linker</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<tr>
<td>microcapillaries</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>
<td>&nbsp;</td>		</tbody>
		</table>
		</div>

making gynogenetic diploid zebrafish by early pressure

二倍体真核生物的杂合性，往往使繁琐的遗传研究。从杂合子女性产生直接可行的纯合二倍体后代的方法，让有害突变，直接在加速向前遗传屏幕的F1诱变女性进行甄别。 Streisinger等人。<sup&gt; 1,2</sup&gt;描述的方法，使雌核发育二倍体（合子）的斑马鱼，斑马鱼的卵用紫外线灭活精子激活使用物理疗法（热或压力），deploymerize微管和防止第二次减数分裂或第一合子细胞分裂。 “初压（EP）的方法将阻塞在减数分裂II，它在受精后不久发生。 EP方法产生一个可行的胚胎的比例较高，可以发展到肥沃的成年人，雌雄兼用。该方法生成的胚胎，除了那些在减数分裂一纯合突变是在EP离合器之间的着丝粒基因位点的50％和小于1％的端粒基因位点检测，通过重组，从他们的着丝粒分离所有位点纯合子。这种方法是复制逐字从斑马鱼书<sup&gt; 3</sup&gt;。

Watch this Scientific Journal Video about Making Gynogenetic Diploid Zebrafish by Early Pressure at JoVE.com

Making Gynogenetic Diploid Zebrafish by Early Pressure

这是一个产生阻塞后，立即用紫外线灭活精子受精的第二次减数分裂雌核发育二倍体斑马鱼的胚胎（仅来自母亲的遗传贡献的胚胎）的方法。 EP胚胎是不完全纯合的，由于在第一次减数分裂重组，但他们是在所有尚未从他们的着丝粒分离，通过重组位点纯合子。

初压雌核发育二倍体的斑马鱼

Heterozygosity in diploid eukaryotes often makes genetic studies cumbersome. Methods that produce viable homozygous diploid offspring directly from ...

Research

JoVE Journal

Biology

Live Imaging of the Zebrafish Embryonic Brain by Confocal Microscopy

现场共聚焦显微镜对斑马鱼的胚胎脑成像

In this video, we demonstrate the method our lab has developed to analyze the cell shape changes and rearrangements required to bend and fold the ...

科研

生物学

Brain Banking: Making the Most of your Research Specimens

脑银行业：使你的研究样本中，大部分

Unbiased stereology is a method for accurately and efficiently estimating the total neuron number (or other cell type) in a given area of interest1.  To ...

Measuring Blood Pressure in Mice using Volume Pressure Recording, a Tail-cuff Method

使用卷压力记录，一条尾巴袖套法小鼠的血压测量

The CODA 8-Channel High Throughput Non-Invasive Blood Pressure system measures the blood pressure in up to 8 mice or rats simultaneously. The CODA ...

Making MR Imaging Child's Play - Pediatric Neuroimaging Protocol, Guidelines and Procedure

使磁共振成像儿戏 - 小儿神经影像学的协议，指引及程序

Within the last decade there has been an increase in the use of structural and functional magnetic resonance imaging (fMRI) to investigate the neural ...

Generating Chimeric Zebrafish Embryos by Transplantation

移植嵌合斑马鱼胚胎生成

One of the most powerful tools used to gain insight into complex developmental processes is the analysis of chimeric embryos. A chimera is defined as an ...

Atmospheric-pressure Molecular Imaging of Biological Tissues and Biofilms by LAESI Mass Spectrometry

常压LAESI质谱生物组织和生物膜的分子成像

Ambient ionization methods in mass spectrometry allow analytical investigations to be performed directly on a tissue or biofilm under native-like ...

Direct Analysis of Single Cells by Mass Spectrometry at Atmospheric Pressure

在大气压力质谱直接分析单细胞

Analysis of biochemicals in single cells is important for understanding cell metabolism, cell cycle, adaptation, disease states, etc. Even the same cell ...

In vivo Electroporation of Morpholinos into the Adult Zebrafish Retina

In vivo Electroporation of Morpholinos into the Adult Zebrafish Retina

在体内电穿孔Morpholinos到成年斑马鱼视网膜

Many devastating inherited eye diseases result in progressive and irreversible blindness because humans cannot regenerate dying or diseased retinal ...

In Vivo 4-Dimensional Tracking of Hematopoietic Stem and Progenitor Cells in Adult Mouse Calvarial Bone Marrow

In Vivo 4-Dimensional Tracking of Hematopoietic Stem and Progenitor Cells in Adult Mouse Calvarial Bone Marrow

在造血干细胞和祖细胞的成年小鼠颅骨骨髓体内四维跟踪

Through a delicate balance between quiescence and proliferation, self renewal and production of differentiated progeny, hematopoietic stem cells (HSCs) ...

Production of Haploid Zebrafish Embryos by In Vitro Fertilization

Production of Haploid Zebrafish Embryos by In Vitro Fertilization

生产单倍体斑马鱼的胚胎通过<em&gt;体外</em&gt;施肥

The zebrafish has become a mainstream vertebrate model that is relevant for many disciplines of scientific study. Zebrafish are especially well suited for ...