Name: imaging calcium responses in gfp-tagged neurons of hypothalamic mouse brain slices
Uploaded: 2012-08-24T13:00:00
Description: Watch this Scientific Journal Video about Imaging Calcium Responses in GFP-tagged Neurons of Hypothalamic Mouse Brain Slices at JoVE.com

我们感谢我们的同事参加工作的总结。这项工作是支持的补助金从德意志研究联合会（SFB 894），的DFG Schwerpunktprogramm 1392“综合分析的嗅觉和德国大众汽车基金会（TLZ）。 TLZ是一个利希滕贝格教授，德国大众汽车基金会。

1。解决方案和琼脂糖凝胶的制备<ol><li>准备所述细胞外液用双蒸水的表。的pH值将是〜7.3后，10分钟曝气Carbogen增（95％O 2/5％CO 2），300毫渗8的渗透压。如果需要更高的渗透压是必需的，它可以进行调整，通过添加更多的葡萄糖（1mM的等于1毫渗）。该溶液被过滤两次，使用0.2μm的膜过滤器，以消除灰尘颗粒和可能的细菌污染。 </li></ol><table border="1"><tbody><tr><td> 试剂名称</stro...

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在神经科学中的一个主要问题是了解大脑是如何处理社会信息化。一个主要的编码嗅觉或信息素信号源所需的信息社会的认可。这些信号在鼻子和识别的信号在大脑中，尤其是下丘脑的神经元群体的检测，发挥了关键作用，许多社会进程和影响的激素和其他神经内分泌因子13-16。的一个重要障碍等脑区的神经元的反应，下丘脑多功能与多个神经元细胞核的分析是感兴趣的特定神经元的鉴定...

There was a typo in the abstract of <a href="http://www.jove.com/video/4213">Imaging Calcium Responses in GFP-tagged Neurons of Hypothalamic Mouse Brain Slices</a>.

Erratum: Imaging Calcium Responses in GFP-tagged Neurons of Hypothalamic Mouse Brain Slices

<table border="1"><tbody><tr><td> 名称 </td><td> 公司 </td><td> 猫。 N° </td></tr><tr><td>洋菜</td><td>西格玛</td><td> A1296 </td></tr><tr><td> Fura-red/AM </td><td> Invitrogen公司</td><td> F-3021 </td></tr><tr><td>聚醚F-127 </td><td>西格玛</td><td> P2443 </td></tr><tr><td>二甲基亚砜</td><td> Fisher Scientific则</td><td> BP231 </td></tr><tr><td>振动刀片切片岩狸V 50 </td><td>蔡司</td><td> 9770170 </td></tr><tr><td>冷却装置CU切片岩狸V 50 65 </td><td>蔡司</td><td> 9920120 </td></tr><tr><td> O 2 / CO 2孵化器，CB210-UL </td><td>宾德</td><td> 0019389 </td></tr><tr><td>超级胶水，乐泰406TM </td><td> ·亨克尔</td><td> 142580 </td></tr><tr><td>双铲，勺形</td><td>Bochem </td><td> 3182 </td></tr><tr><td> Microspoon铲，勺形</td><td> Bochem </td><td> 3344 </td></tr><tr><td>春风似剪刀，莫里亚Vannas-沃尔夫 - 7毫米的刀片</td><td>精细科学工具</td><td> 15370-52 </td></tr><tr><td>春风似剪刀，维纳斯 - 3毫米的刀片</td><td>精细科学工具</td><td> 15000-00 </td></tr><tr><td>瓦格纳剪刀</td><td>精细科学工具</td><td> 14071-12 </td></tr><tr><td>医疗钳，杜蒙7B </td><td>精细科学工具</td><td> 11270-20 </td></tr><tr><td>大矩形的开放式沐浴室（RC-27） </td><td>华纳仪器</td><td> 64-0238 </td></tr><tr><td>共聚焦显微镜BioRad公司光辉2100 </td><td>蔡司</td><td>不适用</td></tr></tbody></table>

imaging calcium responses in gfp-tagged neurons of hypothalamic mouse brain slices

在我们的知识尽管一个巨大的增加有关的机制，在大脑中的信息的编码，一个中心问题的确切分子步骤，以及特定的神经元的活性，在如下丘脑保持脑区的多功能核。这个问题包括参与各种神经激素信号转导通路的调节成分的分子识别。立面图细胞内Ca 2 +在调节神经元的敏感性，发挥重要的作用，无论是在水平的信号转导，和在突触站点。 出现了新的工具，以帮助识别特定的启动子的控制下表达绿色荧光蛋白（GFP）无数的神经元在大脑的神经元。若要监视空间和时间上的刺激诱 ​​导的Ca 2 +的响应在GFP-标记的神经元，非绿色荧光的Ca 2 +指示剂染料Ň要使用的电火工品。此外，激光共聚焦显微镜是一个喜爱的成像单个神经元的方法，由于其能力，可视化神经元的组织内，并在不同的平面的深度限制满分的聚焦荧光组织切片。比例式的Ca 2 +指示剂fura-2的组合已被用于与GFP标记的神经元1。然而，染料通过紫外线（UV）光被激发。 UV光的激光和光穿透深度有限的成本阻碍了它的使用在许多实验室中。此外，GFP荧光可能会干扰与fura-2的信号2。因此，我们决定使用红色荧光的Ca 2 +指示剂染料。巨大的笔画呋喃红色移位允许使用一个单一的激发波长与GFP组合的红色荧光的多色分析。我们有以前了良好的效果，使用呋喃结合红色与绿色荧光蛋白标记的嗅觉神经元。嗅觉组织切片的协议似乎工作电子邮件qually下丘脑神经元。呋喃红色的Ca 2 +成像也成功地结合GFP标记的胰岛β细胞和GFP标记的受体表达的HEK细胞5,6。一个小怪癖的呋喃红是在650 nm，其荧光强度降低，一旦指标结合钙7。因此，具有相对高的静息神经元低的Ca 2 +浓度的荧光强度。应当指出，红色，其他Ca 2 +的指示剂染料存在的或正在开发的，这可能给在不同的神经元和大脑区域更好的或改进的结果。

Watch this Scientific Journal Video about Imaging Calcium Responses in GFP-tagged Neurons of Hypothalamic Mouse Brain Slices at JoVE.com

Imaging Calcium Responses in GFP-tagged Neurons of Hypothalamic Mouse Brain Slices

在这个协议中，我们更新了最新进展，成像钙<sup&gt; 2 +</sup&gt;绿色荧光蛋白标记的神经元在脑组织切片，使用红色荧光钙信号<sup&gt; 2 +</sup&gt;指示剂染料。

在GFP标记的下丘脑神经元的小鼠脑片成像钙反应

Despite an enormous increase in our knowledge about the mechanisms underlying the encoding of information in the brain, a central question concerning the ...

Research

JoVE Journal

Neuroscience

Dual Somatic Recordings from Gonadotropin-Releasing Hormone (GnRH) Neurons Identified by Green Fluorescent Protein (GFP) in Hypothalamic Slices

Dual Somatic Recordings from Gonadotropin-Releasing Hormone GnRH Neurons Identified by Green Fluorescent Protein GFP in Hypothalamic Slices

双体录音从促性腺激素释放激素（GnRH）神经元下丘脑薄片鉴定绿色荧光蛋白（GFP）

Gonadotropin-Releasing Hormone (GnRH) is a small neuropeptide that regulates pituitary release of luteinizing hormone (LH) and follicle-stimulating ...

科研

神经科学

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 ...

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 ...

In vivo Postnatal Electroporation and Time-lapse Imaging of Neuroblast Migration in Mouse Acute Brain Slices

In vivo Postnatal Electroporation and Time-lapse Imaging of Neuroblast Migration in Mouse Acute Brain Slices

成神经细胞迁移的小鼠急性脑片在体内电穿孔产后和时间推移成像

The subventricular zone (SVZ) is one of the main neurogenic niches in the postnatal brain. Here, neural progenitors proliferate and give rise to ...

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+ ...

Ratiometric Calcium Imaging of Individual Neurons in Behaving Caenorhabditis Elegans

Ratiometric Calcium Imaging of Individual Neurons in Behaving Caenorhabditis Elegans

比率钙显像对单个神经元的行为线虫线虫

It has become increasingly clear that neural circuit activity in behaving animals differs substantially from that seen in anesthetized or immobilized ...

Ex Vivo Calcium Imaging for Visualizing Brain Responses to Endocrine Signaling in Drosophila

Ex Vivo Calcium Imaging for Visualizing Brain Responses to Endocrine Signaling in Drosophila

前体钙成像用于可视化大脑对果蝇内分泌信号的反应

Organ-to-organ communication by endocrine signaling, for example, from the periphery to the brain, is essential for maintaining homeostasis. As a model ...

Ballistic Labeling of Pyramidal Neurons in Brain Slices and in Primary Cell Culture

脑切片和原细胞培养中金字塔神经元的弹道标记

It has been reported that the size and shape of dendritic spines is related to their structural plasticity. To identify the morphological structure of ...

In vivo Calcium Imaging of Mouse Geniculate Ganglion Neuron Responses to Taste Stimuli

在活体钙成像小鼠致病性甘里昂神经元对味觉刺激的反应

Within the last ten years, advances in genetically encoded calcium indicators (GECIs) have promoted a revolution in in vivo functional imaging. Using ...

In vivo Calcium Imaging in Mouse Inferior Olive

In vivo Calcium Imaging in Mouse Inferior Olive

在体内 小鼠劣质橄榄中的钙成像

Inferior olive (IO), a nucleus in the ventral medulla, is the only source of climbing fibers that form one of the two input pathways entering the ...