Name: preparation of horizontal slices of adult mouse retina for electrophysiological studies
Uploaded: 2017-01-27T14:00:00
Description: Watch this Scientific Journal Video about Preparation of Horizontal Slices of Adult Mouse Retina for Electrophysiological Studies at JoVE.com

This work was supported by the Deutsche Forschungsgemeinschaft DFG FOR701 and FE464/11-1 to A.F. The authors want to acknowledge the excellent technical assistance of Andrea Nerz.

所有程序都根据用于由德国联邦共和国发出动物实验的准则进行。 注意：由于视网膜组织将在此过程中的延长部分是无氧，所有步骤应尽可能快地进行。小鼠应该暗适应解剖前至少3小时。以避免视网膜，制备的光适应和切片应在昏暗的红光下进行。 1.媒体和其他化学品的制备<ol><li> 准备艾姆斯介质6。解剖后，沉浸视网膜组织埃...

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在对比垂直切片，如水平细胞和无长突细胞径向定向神经元的树突形态保存完好。相比于整个视网膜的准备工作，这些细胞类型是微电极方便且可由两个电生理和成像技术进行研究。在这里，我们专注于视网膜外层;然而，类似的方法已经报道尤其用于内视网膜10的无长突细胞。 水平切片准备的属性有功能性研究的几个问题。锥感光细胞和突触后水平细胞之?...

视觉信息被编码为感光体活化的动态空间 - 时间模式，及光感受器和二阶神经元之间的色带突触确定的视觉信息的下行传输。哺乳动物视网膜的垂直切片制备是高度有价值的工具来研究在垂直通路的信号处理， 即感光细胞和双极细胞之间，和之间双极细胞和某些类型的无长突细胞1，2，3，4。 然而，垂直切片总是导致许多视网膜神经元的树突领域的严重截断，导致突触联系的相当大的损失。特别是在外侧视网膜，水平细胞由于它们的横向扩展的扩展树突字段和轴突终端系统5的影响。在垂直因而切片制备，数百锥感光体终端到水平细胞的树突的突触输入被降低到几稀疏联系人。这可能足以研究单个突触的性质，但是它确实绝非表示感光体色带突触的同步激活基础的信号处理能力。 因此，我们开发了一个横向切片准备，这让几乎所有的锥感光的突触输入水平细胞树突的完整和功能。平行于视网膜表面切片运行的平面上，通过水平细胞和无长突细胞之间的内核层理想地切割。因此，外视网膜的水平片段创建含有上突触前部位，用内外段以及突触终端，水平细胞和双极细胞上的突触后现场光感受器。这种准备是非常适合ŤØ通过其树突领域内的所有锥感光细胞研究协调突触的投入水平细胞树突。它因此可能证明是有用的，以便更好地理解在感光色带突触，这是视觉信息的传送从感光体活化的基质成突触后响应关键的运作。

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			<td height="21" style="height:21px;width:216px;">Ames&#39; Medium</td>
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			<td style="width:119px;">A1420</td>
			<td style="width:167px;"></td>
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			<td height="21" style="height:21px;width:216px;">Sodium Bicarbonate</td>
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			<td style="width:119px;">S5761</td>
			<td></td>
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		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Hepes</td>
			<td style="width:125px;">Sigma</td>
			<td style="width:119px;">H3375</td>
			<td></td>
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		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Carbogen</td>
			<td style="width:125px;">Air Liquide</td>
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			<td height="21" style="height:21px;width:216px;">Agarose</td>
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			<td style="width:119px;">A9045</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">35-mm petri dish (plastic)</td>
			<td style="width:125px;">Nunc Thermofisher</td>
			<td style="width:119px;">150318</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Glass petri dish</td>
			<td style="width:125px;">Chemoline</td>
			<td style="width:119px;">50-1470-40</td>
			<td></td>
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		<tr height="63">
			<td height="63" style="height:63px;width:216px;">Isoflurane</td>
			<td style="width:125px;">Dispensary university hospital</td>
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			<td height="21" style="height:21px;width:216px;">Vibratome injector blade</td>
			<td style="width:125px;">Biosystems</td>
			<td style="width:119px;">39053250</td>
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		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Vibratome</td>
			<td style="width:125px;">Leica Microsystems</td>
			<td style="width:119px;">VT1200</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Vibrocheck</td>
			<td style="width:125px;">Leica Microsystems</td>
			<td style="width:119px;"></td>
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			<td height="21" style="height:21px;width:216px;">Microwave</td>
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			<td style="width:119px;"></td>
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			<td height="21" style="height:21px;width:216px;">Water bath</td>
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			<td height="21" style="height:21px;width:216px;">Forceps</td>
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			<td height="21" style="height:21px;width:216px;">20-gauge needles</td>
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		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Spring scissors</td>
			<td style="width:125px;"></td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Curved scalpel blades</td>
			<td style="width:125px;"></td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Stereo microscope</td>
			<td style="width:125px;"></td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Plastic Pasteur pipette</td>
			<td style="width:125px;"></td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Glass Pasteur pipette</td>
			<td style="width:125px;"></td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
	</tbody>
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preparation of horizontal slices of adult mouse retina for electrophysiological studies

垂直切片准备工作已经非常成熟，研究电路和信号传输在成年哺乳动物的视网膜。这些制剂切片的平面垂直于视网膜表面，使其成为理想的像光感受器和双极细胞径向定向神经元的研究。然而，水平细胞，宽视场的无长突细胞和神经节细胞的大树突乔木大多截短，留下显着降低突触活动在这些细胞中。而神经节细胞和无长突移位细胞可以在视网膜的一个整体安装的制剂进行研究，水平细胞和位于内核层无长突细胞是只很差在整个视网膜组织电极访问。 为了实现最方便的访问和突触的完整性，我们开发了小鼠视网膜的水平切片准备，并就读于光感受器和水平之间的突触信号传递细胞。横切片允许（1）容易和明确为电极定位水平细胞体的视觉识别，和（2）保存在扩展水平细胞树突的字段，作为完整且功能锥体突触输入到水平细胞树突的先决条件。 从水平切片水平细胞表现出补品突触活动在黑暗中，他们回答了介绍光闪烁以减少内向电流和减弱突触的活动。免疫细胞化学证据表明，水平细胞的树突领域内几乎所有的锥体建立与周边树突突触。因此，水平片段制备是公适合研究水平延伸视网膜神经元的跨越选定突触生理特性以及传感信号传输和整合。

<html>在小鼠视网膜的水平片段，水平细胞体可以很容易地根据它们与几个主要树突从多边形细胞体（ 图1A）的新兴独特的形态识别。位于切片的表面附近水平细胞机构是对膜片钳电极例行访问。期间记录，将细胞填充有荧光染料，揭示其复杂树突分支，这非常类似于完整小鼠视网膜8（ 图1B）的轴突承载水平细胞的形态。用荧光?...

Watch this Scientific Journal Video about Preparation of Horizontal Slices of Adult Mouse Retina for Electrophysiological Studies at JoVE.com

Preparation of Horizontal Slices of Adult Mouse Retina for Electrophysiological Studies

我们开发了成年哺乳动物视网膜的水平切片准备具有平行切片于视网膜表面的平面。非径向导向的视网膜神经元树突领域并没有截断，使信号处理与膜片钳和成像技术研究。

成年小鼠视网膜电生理研究水平切片的制备

Vertical slice preparations are well established to study circuitry and signal transmission in the adult mammalian retina. The plane of sectioning in ...

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Neuroscience

Organotypic Culture of Full-thickness Adult Porcine Retina

器官型培养的全厚成年猪视网膜

There is a recognized demand for in vitro models that can replace or reduce animal experiments. Porcine retina has a similar neuronal structure to human ...

科研

神经科学

In vivo Electroporation of Developing Mouse Retina

In vivo Electroporation of Developing Mouse Retina

在发展中国家小鼠视网膜体内电穿孔

The functional characterization of genes expressed during mammalian retinal development remains a significant challenge.  Gene targeting to generate ...

Electrophysiological Characterization of GFP-Expressing Cell Populations in the Intact Retina

在完整的视网膜电生理特性表达GFP的细胞群

Studying the physiological properties and synaptic connections of specific neurons in the intact tissue is a challenge for those cells that lack ...

Time-lapse Imaging of Neuroblast Migration in Acute Slices of the Adult Mouse Forebrain

急性片的成年小鼠前脑的神经细胞迁移的延时成像

There is a substantial body of evidence indicating that new functional neurons are constitutively generated from an endogenous pool of neural stem cells ...

Preparation of Acute Subventricular Zone Slices for Calcium Imaging

钙成像的急性下区片的制备

The subventricular zone (SVZ) is one of the two neurogenic zones in the postnatal brain. The SVZ contains densely packed cells, including neural ...

Dual Electrophysiological Recordings of Synaptically-evoked Astroglial and Neuronal Responses in Acute Hippocampal Slices

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Astrocytes form together with neurons tripartite synapses, where they integrate and modulate neuronal activity. Indeed, astrocytes sense neuronal inputs ...

Improved Preparation and Preservation of Hippocampal Mouse Slices for a Very Stable and Reproducible Recording of Long-term Potentiation

长时程增强一个非常稳定的和可重复的记录和保存海马鼠标片制备工艺的改进

Long-term potentiation (LTP) is a type of synaptic plasticity characterized by an increase in synaptic strength and believed to be involved in memory ...

Electrophysiological Recording in the Brain of Intact Adult Zebrafish

在完整的成年斑马鱼大脑电生理记录

Previously, electrophysiological studies in adult zebrafish have been limited to slice preparations or to eye cup preparations and electrorentinogram ...

Healthy Brain-pituitary Slices for Electrophysiological Investigations of Pituitary Cells in Teleost Fish

健康脑-垂体切片对硬骨鱼类鱼垂体细胞电生理的研究

Electrophysiological investigations of pituitary cells have been conducted in numerous vertebrate species, but very few in teleost fish. Among these, the ...

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成人脑切片长期抑郁症诱导评估

Synaptic plasticity provides a mechanism for learning and memory. For cerebellar motor learning, long-term depression (LTD) of synaptic transmissions from ...