Name: gross and fine dissection of inner ear sensory epithelia in adult zebrafish (danio rerio)
Uploaded: 2009-05-08T00:00:00
Description: Watch this Scientific Journal Video about Gross and Fine Dissection of Inner Ear Sensory Epithelia in Adult Zebrafish Danio rerio at JoVE.com

这项研究是由国家人类基因组研究所，美国国立卫生研究院院内研究计划的支持。

第1部分：组织准备<ol><li>安乐死的成年斑马鱼缓冲的MS - 222（甲酸乙酯3 - 氨基磺酸盐，0.03％）。 </li><li>杀头的鱼。切断下颌，头部腹侧的夹层垫，带虫针固定的组织。取出头骨腹软组织和一双细镊子显微镜下解剖，打开颅骨囊腹侧部分。 </li><li>鱼头中的多聚甲醛（4％），固定在4 ° C过夜。 </li><li>在PBS冲洗固定鱼嘴（磷酸盐缓冲液，1X）3次，每次5分钟。 </li></ol>第2部分：内耳?...

在这个协议中，我们描述的内耳感觉上皮细胞的成年斑马鱼，研究的感官上皮细胞和亚细胞的功能，使我们准备组织样本的总罚款清扫。 为了开展一个成功的解剖，第一步，固定的，是非常重要的。确保您始终使用新鲜配制的多聚甲醛溶液，因为旧的解决方案将导致是很难处理过程中剥离的组织下，固定。但是，请确保您不要过度修复组织，由他们留在固定时间过长的解决?...

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		<tr>
		<th>Material Name</th>
		<th>Type</th>
		<th>Company</th>
		<th>Catalogue Number</th>
		<th>Comment</th>
		</tr>
		</thead>
		<tbody>
		
<tr>
<td>MS222 (Ethyl 3-aminobenzoate methanesulfonate salt)</td>
<td>Reagent</td>
<td>Sigma</td>
<td>A5040</td>
<td>&nbsp;</td>
<tr>
<td>PBS (Phosphate-buffered Saline, 10X)</td>
<td>Reagent</td>
<td>Mediatech</td>
<td>46-013-CM</td>
<td>&nbsp;</td>
<tr>
<td>Paraformaldehyde</td>
<td>Reagent</td>
<td>Sigma</td>
<td>P-6148</td>
<td>&nbsp;</td>
<tr>
<td>Triton X-100</td>
<td>Reagent</td>
<td>Sigma</td>
<td>T8787</td>
<td>&nbsp;</td>
<tr>
<td>Alexa Fluor 488 phalloidin</td>
<td>Reagent</td>
<td>Invitrogen</td>
<td>A12379</td>
<td>&nbsp;</td>
<tr>
<td>VECTASHIELD Hard Set Mounting Medium with DAPI</td>
<td>Reagent</td>
<td>Vector Laboratories</td>
<td>H-1500</td>
<td>&nbsp;</td>
<tr>
<td>Dissection microscope</td>
<td>Microscope</td>
<td>Nikon</td>
<td>SMZ1000</td>
<td>&nbsp;</td>
<tr>
<td>Insect pins</td>
<td>Tool</td>
<td>Fine Science Tools</td>
<td>26000-40</td>
<td>&nbsp;</td>
<tr>
<td>Scissors</td>
<td>Tool</td>
<td>Roboz</td>
<td>RS-5880</td>
<td>&nbsp;</td>
<tr>
<td>Tweezers</td>
<td>Tool</td>
<td>Roboz</td>
<td>RS-5045</td>
<td>&nbsp;</td>
<tr>
<td>Needle blade</td>
<td>Tool</td>
<td>Sharpoint</td>
<td>78-6810</td>
<td>&nbsp;</td>
<tr>
<td>Fluorescent microscope</td>
<td>Microscope</td>
<td>Zeiss</td>
<td>Axiovert 200M</td>
<td>&nbsp;</td>
<tr>
<td>Dissection pad</td>
<td>Tool</td>
<td>eNasco</td>
<td>SB07840M</td>
<td>&nbsp;</td>
<tr>
<td>Zebrafish</td>
<td>Animal</td>
<td>various</td>
<td>n/a</td>
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gross and fine dissection of inner ear sensory epithelia in adult zebrafish (danio rerio)

感觉神经上皮细胞在内耳的听觉和平衡功能的关键结构。因此，重要的是了解，这主要是两种类型的细胞组成的上皮细胞和分子特征：毛细胞（HCS）和支持细胞（SCS）。在这里，我们选择研究的内耳感觉上皮细胞在成年斑马鱼不仅的上皮结构，因为在所有研究的脊椎动物是高度保守的，但也因为成年斑马鱼能够再生HCS，有能力的哺乳动物出生后失去不久。作为一个模型系统，研究可能有助于为临床治疗人类慧聪损失的听觉/平衡赤字的成年脊椎动物内耳HC再生机制，我们使用的成年斑马鱼的内耳。 在这里，我们将演示如何在成年斑马鱼的内耳感觉上皮的毛保费及精细解剖。总清扫删除周围的内耳组织，并准备组织切片，这使得我们可以研究的感觉上皮细胞的详细结构。罚款清扫清理每一个人的上皮细胞的非感觉上皮组织，使我们能够在整个安装的上皮细胞样本研究整个上皮细胞的异质性很容易。

Watch this Scientific Journal Video about Gross and Fine Dissection of Inner Ear Sensory Epithelia in Adult Zebrafish Danio rerio at JoVE.com

Gross and Fine Dissection of Inner Ear Sensory Epithelia in Adult Zebrafish Danio rerio

成年斑马鱼的内耳感觉上皮是一个良好的模型系统，为了解在成年脊椎动物的毛细胞再生的机制。此协议表明的上皮细胞的精细解剖，我们可以通过它获得组织样本，研究在细胞和亚细胞水平的再生事件。

格罗斯和精细解剖内耳感觉上皮细胞在成年斑马鱼（斑马鱼）

Neurosensory epithelia in the inner ear are the crucial structures for hearing and balance functions. Therefore, it is important to understand the ...

Research

JoVE Journal

Biology

Gross and Fine Dissection of Inner Ear Sensory Epithelia in Adult Zebrafish (Danio rerio)

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The stomach of the American lobster (Homarus americanus) is located in the cephalothorax, between the rostrum and the cervical groove. The anterior end of ...

科研

生物学

Dissection of Organs from the Adult Zebrafish

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Over the last 20 years, the zebrafish has become a powerful model organism for understanding vertebrate development and disease.  Although experimental ...

Direct Delivery of MIF Morpholinos Into the Zebrafish Otocyst by Injection and Electroporation Affects Inner Ear Development

通过注射和电穿孔的斑马鱼Otocyst的MIF Morpholinos直接运抵影响内耳发展

In recent years, electroporation has become a popular technique for in vivo transfection of DNA, RNA, and morpholinos into various tissues, including the ...

Dissection of the Adult Zebrafish Kidney

成年斑马鱼肾脏的解剖

Researchers working in the burgeoning field of adult stem cell biology seek to understand the signals that regulate the behavior and function of stem ...

Heart Dissection in Larval, Juvenile and Adult Zebrafish, Danio rerio

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幼虫，少年和成年斑马鱼的心脏解剖，斑马鱼</em

Zebrafish have become a beneficial and practical model organism for the study of embryonic heart development (see recent reviews1-6), however, work ...

In vivo Electroporation of Morpholinos into the Adult Zebrafish Retina

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

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定期保养和维护的斑马鱼（<em&gt;斑马鱼</em&gt;）实验室简介

This protocol describes regular care and maintenance of a zebrafish laboratory. Zebrafish are now gaining popularity in genetics, pharmacological and ...

In Vivo Modeling of the Morbid Human Genome using Danio rerio

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在体内病态的人类基因组建模使用斑马鱼

Here,  we present methods for the development of assays to query potentially clinically  significant nonsynonymous changes using in  vivo complementation ...

Gavaging Adult Zebrafish

Gavaging成年斑马鱼

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

High Throughput Danio Rerio Energy Expenditure Assay

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高通量<em&gt;斑马鱼</em&gt;能量消耗分析

Zebrafish are an important model organism with inherent advantages that have the potential to make zebrafish a widely applied model for the study of ...