Name: imaging neuronal responses in slice preparations of vomeronasal organ expressing a genetically encoded calcium sensor
Uploaded: 2011-12-06T13:00:00
Description: Watch this Scientific Journal Video about Imaging Neuronal Responses in Slice Preparations of Vomeronasal Organ Expressing a Genetically Encoded Calcium Sensor at JoVE.com

我们感谢安德烈莫兰连同实验动物服务设施（LASF）的成员，他们对畜牧业和技术服务的优秀支持在斯托瓦斯研究所。这项工作是支持，从斯托瓦斯研究所和国立卫生研究院（NIDCD 008003）资金，哭了。内容完全是作者的责任，并不一定代表美国国立耳聋与其他交流障碍研究所或国立卫生研究院的官方意见。美国正在申请专利的斯托瓦斯研究所，哭​​和LM的tetO - G CaMP2小鼠。

1。溶液的配制<ol><li>根据下表，准备R1的10X，10X的R2和10X R3的解决方案。 R1 <table border="1"><tbody><tr><td>化学品</td><td>兆瓦（克/摩尔） </td><td>毫米（1X） </td><td> 10倍的股票（G / L） </td></tr><tr><td>氯化钠</td><td> 58.44 </td><td> 125 </td><td> 73.05 </td></tr><tr><td>氯化钾</td><td> 74.55 </td><td> 2.5 </td><td> 1.86 </td></tr><tr><td> MgCl 2的 </td><td> 1中号股票</td><td> 1 </td><td> 10毫升</td></tr><tr><td>氯化钙2 ...

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大多数的犁鼻器受体（VRS）保持作为，因为他们发现由杜拉克和Axel 5的孤儿受体。这些化学感应受体和他们在调解动物行为中的作用的信息素配体不能很好地理解。到现在为止，只有一对配体/受体，ESP1肽及其受体，Vmn2r116（V2Rp5），已确定并传达特定的社会信息，19,23。另一个受体，V1rb2，已被证明回应2 -庚酮，这大概是从雌性小鼠24尿液丰富。然而，此配体所传达的具体?...

imaging neuronal responses in slice preparations of vomeronasal organ expressing a genetically encoded calcium sensor

犁鼻器（VNO）检测，化学感应信号在同一物种的1,2个人的社会，性和生殖状态信息。这些种内的信号，信息素，以及来自一些食肉动物3的信号，激活高水平的特异性和灵敏度 4犁鼻感觉神经元（VSNS） 。 G蛋白偶联受体V1R，V2R和FPR 5-14，至少有三个不同的家庭表示在犁鼻神经元的调解检测的化学感应线索。为了了解信息素的信息是如何编码的VNO，它是分析个人VSNS响应各种刺激，并确定这些反应的特异性受体介导的关键。 VNO neuroepithelia括在一对犁骨头。 VNO半盲管状结构有一个开口端（犁鼻管）连接到鼻腔。 VSNS延长树突的犁鼻器信息素线索接触表达的受体在树突状旋钮管腔部分。 VSNS的细胞体的形式与V1R和V2R伪分层表示层分别6-8在根尖和基底层。几项技术已被用于监察VSNS 4,12,15-19感官刺激的反应。急性切片准备在这些技术中，有几个优点。首先，3,17分离VSNS相比，切片的筹备工作保持在其原生形态和细胞树突的神经元保持相对完好。二，VSNS胞体很容易在冠状片的VNO访问，允许电生理研究和成像实验相比，整个上皮细胞和整个安装准备工作 12,20 。第三，这种方法可以结合分子克隆技术，让受体识别。感官刺激会引起强烈的Ca 2 + VSNS涌入，表明受体激活 4,21 。因此，我们开发的转基因小鼠的嗅感觉神经元，其中包括VSNS 15,22表达的G - CaMP2。探针的灵敏度和遗传性，极大地促进钙离子成像实验。这种方法消除了染料4,21以前的研究中使用的加载过程。我们还采用了配体传递系统，可应用各种刺激的VNO片。这两种技术的结合使我们能够监测多个神经元，同时响应大量的刺激。最后，我们已经建立了一个半自动化分析的管道，以协助图像处理。

Watch this Scientific Journal Video about Imaging Neuronal Responses in Slice Preparations of Vomeronasal Organ Expressing a Genetically Encoded Calcium Sensor at JoVE.com

Imaging Neuronal Responses in Slice Preparations of Vomeronasal Organ Expressing a Genetically Encoded Calcium Sensor

犁鼻器（VNO）检测种内的化学信号，传达社会和生殖信息。我们已经进行钙离子成像实验，利用转基因小鼠犁鼻组织中表达的G - CaMP2。这种方法可以让我们来分析复杂的犁鼻器神经元的反应模式，以大量的信息素的刺激。

在犁鼻器官的切片准备工作表示一个基因编码的钙传感器成像神经元的反应

The vomeronasal organ (VNO) detects chemosensory signals that carry information about the social, sexual and reproductive status of the individuals within ...

Research

JoVE Journal

Neuroscience

Calcium Imaging of Odor-evoked Responses in the Drosophila Antennal Lobe

Calcium Imaging of Odor-evoked Responses in the Drosophila Antennal Lobe

在气味诱发反应的钙成像果蝇触角叶

The antennal lobe is the primary olfactory center in the insect brain and represents the anatomical and functional equivalent of the vertebrate olfactory ...

科研

神经科学

Imaging Pheromone Sensing in a Mouse Vomeronasal Acute Tissue Slice Preparation

成像遥感信息素在小鼠犁鼻急性组织切片的制备

Peter Karlson and Martin L&uuml;scher used the term pheromone for the first time in 19591 to describe chemicals used for intra-species communication. ...

In vivo Neuronal Calcium Imaging in C. elegans

In vivo Neuronal Calcium Imaging in C. elegans

C.在体内神经元钙成像技术线虫

The nematode worm C. elegans is an ideal model organism  for relatively simple, low cost neuronal imaging in vivo. Its small transparent body and simple, ...

Ex Vivo Preparations of the Intact Vomeronasal Organ and Accessory Olfactory Bulb

Ex Vivo Preparations of the Intact Vomeronasal Organ and Accessory Olfactory Bulb

完整的犁鼻器和副嗅球的体外制剂

The mouse accessory olfactory system (AOS) is a specialized sensory pathway for detecting nonvolatile social odors, pheromones, and kairomones. The first ...

Imaging Intracellular Ca2+ Signals in Striatal Astrocytes from Adult Mice Using Genetically-encoded Calcium Indicators

Imaging Intracellular Ca2+ Signals in Striatal Astrocytes from Adult Mice Using Genetically-encoded Calcium Indicators

成像细胞内Ca<sup&gt; 2+</sup&gt;从成年小鼠使用基因编码的钙指标纹状体星形胶质细胞信号

Astrocytes display spontaneous intracellular Ca2+ concentration fluctuations ([Ca2+]i) and in several settings respond to neuronal excitation with ...

Recording Light-evoked Postsynaptic Responses in Neurons in Dark-adapted, Mouse Retinal Slice Preparations Using Patch Clamp Techniques

记录光诱发神经元突触后反应在暗适应，小鼠视网膜切片准备利用膜片钳技术

The retina is the gateway to the visual system. To understand visual signal processing mechanisms, we investigate retinal neural network functions. ...

Imaging Membrane Potential with Two Types of Genetically Encoded Fluorescent Voltage Sensors

成像膜电位与基因编码荧光电压传感器的两种类型

Genetically encoded voltage indicators (GEVIs) have improved to the point where they are beginning to be useful for in vivo recordings. While the ultimate ...

Optical Control of a Neuronal Protein Using a Genetically Encoded Unnatural Amino Acid in Neurons

一个神经元蛋白在神经元使用遗传编码的非天然氨基酸的光控制

Photostimulation is a noninvasive way to control biological events with excellent spatial and temporal resolution. New methods are desired to ...

Prolonged Incubation of Acute Neuronal Tissue for Electrophysiology and Calcium-imaging

对于电生理和钙成像急性神经组织的长时间培养

Acute neuronal tissue preparations, brain slices and retinal wholemount, can usually only be maintained for 6 - 8 h following dissection. This limits the ...

Two-photon Calcium Imaging in Neuronal Dendrites in Brain Slices

脑切片神经元树突的双光子钙成像

Calcium (Ca2+) imaging is a powerful tool to investigate the spatiotemporal dynamics of intracellular Ca2+ signals in neuronal dendrites. Ca2+ ...