Name: revealing neural circuit topography in multi-color
Uploaded: 2011-11-14T12:00:00
Description: Watch this Scientific Journal Video about Revealing Neural Circuit Topography in Multi-Color at JoVE.com

这项工作得到了新的启动资金来自叶史瓦大学阿尔伯特爱因斯坦医学院的研究者RVS。

1。示踪剂在体内提供使用无菌手术（第1.1-1.16） 整个过程中，我们建议使用标准的无菌手术技术。这包括使用无菌手套，隔离衣和口罩。工具应进行高压灭菌或使用前彻底清洁的水，然后乙醇。在手术过程中，特别是与动物之间，干净的纸巾碎片珠用热灭菌（无菌250＃18000-45，精细科学的工具）的工具和干燥消毒仪器关闭。此外，至关重要的是，您组织您的工作空间，?...

<ol>
	<li>Apps, R., Hawkes, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cerebellar+cortical+organization:+a+one-map+hypothesis.">Cerebellar cortical organization: a one-map hypothesis.</a> Nat. Rev. Neurosci. 10, 670-681 (2009).</li><li>Grishkat, H. L., Eisenman, L. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Development+of+the+spinocerebellar+projection+in+the+prenatal+mouse.">Development of the spinocerebellar projection in the prenatal mouse.</a> J. Comp. Neurol. 363, 93-108 (1995).</li><li>Mesulam, 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> Tracing neural connections with horseradish peroxidase. , (1982).</li><li>Reeber, S. L., Sillitoe, R. V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Patterned+expression+of+a+cocaine-+and+amphetamine+regulated+transcript+(CART)+peptide+reveals+complex+circuit+topography+in+the+rodent+cerebellar+cortex.">Patterned expression of a cocaine- and amphetamine regulated transcript (CART) peptide reveals complex circuit topography in the rodent cerebellar cortex.</a> Journal of Comparative Neurology. , (2011).</li><li>Reeber, S. L., Gebre, S. A., Sillitoe, R. V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Fluorescence+mapping+of+afferent+topography+in+three+dimensions.">Fluorescence mapping of afferent topography in three dimensions.</a> Brain. Struct. Funct. , (2011).</li><li>Sillitoe, R. V., Stephen, D., Lao, Z., Joyner, A. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Engrailed+homeobox+genes+determine+the+organization+of+Purkinje+cell+sagittal+stripe+gene+expression+in+the+adult+cerebellum.">Engrailed homeobox genes determine the organization of Purkinje cell sagittal stripe gene expression in the adult cerebellum.</a> J. Neurosci. 28, 12150-12162 (2008).</li><li>Sillitoe, R. V., Vogel, M. W., Joyner, A. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Engrailed+homeobox+genes+regulate+establishment+of+the+cerebellar+afferent+circuit+map.">Engrailed homeobox genes regulate establishment of the cerebellar afferent circuit map.</a> J. Neurosci. 30, 10015-10024 (2010).</li><li>Vig, J., Goldowitz, D., Steindler, D. A., Eisenman, L. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Compartmentation+of+the+reeler+cerebellum:+segregation+and+overlap+of+spinocerebellar+and+secondary+vestibulocerebellar+fibers+and+their+target+cells.">Compartmentation of the reeler cerebellum: segregation and overlap of spinocerebellar and secondary vestibulocerebellar fibers and their target cells.</a> Neuroscience. 130, 735-744 (2005).</li><li>Vogel, M. W., Prittie, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Topographic+spinocerebellar+mossy+fiber+projections+are+maintained+in+the+lurcher+mutant.">Topographic spinocerebellar mossy fiber projections are maintained in the lurcher mutant.</a> J. Comp. Neurol. 343, 341-351 (1994).</li><li>Voogd, J., Broere, G., van Rossum, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+medio-lateral+distribution+of+the+spinocerebellar+projection+in+the+anterior+lobe+and+the+simple+lobule+in+the+cat+and+a+comparison+with+some+other+afferent+fibre+systems.">The medio-lateral distribution of the spinocerebellar projection in the anterior lobe and the simple lobule in the cat and a comparison with some other afferent fibre systems.</a> Psychiatr. Neurol. Neurochir. 72, 137-151 (1969).</li><li>Wu, H. S., Sugihara, I., Shinoda, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Projection+patterns+of+single+mossy+fibers+originating+from+the+lateral+reticular+nucleus+in+the+rat+cerebellar+cortex+and+nuclei.">Projection patterns of single mossy fibers originating from the lateral reticular nucleus in the rat cerebellar cortex and nuclei.</a> J. Comp. Neurol. 411, 97-118 (1999).</li></ol>

我们已经描述了手术和技术的成功使用一种新型的荧光为基础的方法，迅速在发展中国家和成年小鼠的神经预测标签的轴突和树突跟踪所需的详细信息。使用WGA - Alexa的我们展示了如何示踪剂和标记可用于图案电路地形分析在高分辨率和在三个方面。 许多示踪剂追踪神经回路，包括霍乱毒素B亚基，生物胞素，Neurobiotin，菜豆leucoagglutinin（PHAL），fluororuby，fluoremerald和荧光金。?...

<table border="1"><tbody><tr><td> 设备/试剂 </td><td> 型号/目录编号 </td><td> 公司 </td></tr><tr><td>珠灭菌器</td><td>型号无菌250猫。 ＃18000-45 </td><td>精细科学工具</td></tr><tr><td> Cauterizer </td><td>猫。 ＃18000-00 </td><td>精细科学工具</td></tr><tr><td>硼硅玻璃毛细管</td><td>猫。 ＃300056 </td><td>哈佛仪器</td></tr><tr><td>双阶段玻璃微管拖轮</td><td>型号001 - PC - 10 </td><td> Narishige </td></tr><tr><td>千分尺注射器</td><td>猫。 ＃GS - 1100 </td><td> Gilmont仪器</td></tr><tr><td>小动物脑立体定位仪</td><td>型号940 - </td><td> Kopf仪器</td></tr><tr><td>电极的机械手</td><td>型号960 </td><td> Kopf仪器</td></tr><tr><td> Vetcare室</td><td>猫。 ＃340508 </td><td>哈佛仪器</td></tr><tr><td>加热垫</td><td>猫。 ＃341241 </td><td>哈佛仪器</td></tr><tr><td>徕卡DFC360 FX相机</td><td> DFC360外汇</td><td>徕卡</td></tr><tr><td>徕卡DFC490相机</td><td> DFC490 </td><td>徕卡</td></tr><tr><td>徕卡DM5500显微镜</td><td> DM5500 </td><td>徕卡</td></tr><tr><td>徕卡DFC3000 FX相机</td><td> DFC3000外汇</td><td>徕卡</td></tr><tr><td>徕卡MZ16 FA显微镜</td><td> MZ16足总杯</td><td>徕卡</td></tr><tr><td> CY3过滤器</td><td>型号＃11600231 </td><td>徕卡</td></tr><tr><td> FITC标记的筛选</td><td>型号＃11513880 </td><td>徕卡</td></tr><tr><td> A4的DAPI /紫外线过滤器</td><td>型号＃11504162 </td><td>徕卡</td></tr><tr><td>麦胚凝集素的Alexa Fluor 488共轭</td><td>猫。 ＃W11261 </td><td> Invitrogen公司</td></tr><tr><td>小麦GER米凝集素的Alexa Fluor 555共轭</td><td>猫。 ＃W32464 </td><td> Invitrogen公司</td></tr></tbody></table>

revealing neural circuit topography in multi-color

被组织成神经回路的功能地形图。为了形象直观地查看复杂的电路架构，我们开发了一个方法多种地形预测全球的图案，以可靠的标签。小脑是一个理想的的模型来研究神经回路的有序安排。例如，房室脊髓小脑长满青苔的纤维组织已经被证明是一个不可缺少的系统研究苔藓纤维图案的。我们最近表明，麦胚凝集素（WGA）的共轭Alexa的555和488可以用来跟踪，在发展中国家和成年小鼠的脊髓小脑苔藓纤维预测（Reeber等。，2011年）。我们发现了三个主要物业，使标签神经预测的WGA Alexa的示踪剂的理想工具。首先，Alexa的荧光团激烈，其亮度wholemount成像允许后直接跟踪。二，WGA - Alexa的示踪剂标签的整个发展中国家和成人神经projectio轨迹NS。第三，WGA - Alexa的示踪剂正在迅速运逆行及顺行方向。在这里，我们详细描述了如何准备示踪剂和其​​他所需的工具，如何进行手术治疗脊髓小脑跟踪和如何以最佳形象追溯到三个方面的预测。综上所述，我们提供了一个一步一步跟踪协议，将破译的组织和功能的地图不仅在小脑，而且在皮质，脑干和脊髓的连通。

Watch this Scientific Journal Video about Revealing Neural Circuit Topography in Multi-Color at JoVE.com

Revealing Neural Circuit Topography in Multi-Color

我们提供了一个实用指南提供示踪剂<em&gt;在体内</em&gt;，并使用作为一个模型系统的脊髓小脑通路证明成功在小鼠神经元电路分析的基本步骤。我们详细描述了我们全方位的跟踪协议，它利用麦胚凝集素（WGA）的共轭Alexa的荧光团。

多色揭示神经回路地形

Neural circuits are organized into functional topographic maps. In order to visualize complex circuit architecture we developed an approach to reliably ...

Research

JoVE Journal

Neuroscience

Dissection and Culture of Commissural Neurons from Embryonic Spinal Cord

从胚胎脊髓​​连合神经的解剖和文化

Commissural neurons have been widely used to investigate the mechanisms underlying axon guidance during embryonic spinal cord development. The cell bodies ...

科研

神经科学

Combining Behavioral Endocrinology and Experimental Economics: Testosterone and Social Decision Making

睾酮和社会决策，结合行为内分泌和实验经济学：

Behavioral endocrinological research in humans as well as in animals suggests that testosterone plays a key role in social interactions. Studies in ...

Time-lapse Live Imaging of Clonally Related Neural Progenitor Cells in the Developing Zebrafish Forebrain

在发展中国家斑马鱼的前脑的神经祖细胞克隆相关的定时实时成像

Precise patterns of division, migration and differentiation of neural progenitor cells are crucial for proper brain development and function1,2. To ...

Detection of Microregional Hypoxia in Mouse Cerebral Cortex by Two-photon Imaging of Endogenous NADH Fluorescence

检测内源NADH的荧光双光子成像的小鼠大脑皮层Microregional缺氧

The brain's ability to function at high levels of metabolic demand depends on continuous oxygen supply through blood flow and tissue oxygen diffusion. ...

Wholemount Immunohistochemistry for Revealing Complex Brain Topography

为揭示复杂的大脑地形wholemount免疫组化

The repeated and well-understood cellular architecture of the cerebellum make it an ideal model system for exploring brain topography. Underlying its ...

Design and Assembly of an Ultra-light Motorized Microdrive for Chronic Neural Recordings in Small Animals

设计和组装的超轻型电动微型硬盘（Microdrive）慢性神经录音的小动物

The ability to chronically record from populations of neurons in freely behaving animals has proven an invaluable tool for dissecting the function of ...

Multi-unit Recording Methods to Characterize Neural Activity in the Locust (Schistocerca Americana) Olfactory Circuits

Multi-unit Recording Methods to Characterize Neural Activity in the Locust Schistocerca Americana Olfactory Circuits

多单位记录方法来描述蝗虫的神经活动（<em&gt; Schistocerca美洲</em&gt;）嗅觉电路

Detection and interpretation of olfactory cues are critical for the survival  of many organisms. Remarkably, species across phyla have strikingly similar ...

Neural Circuit Recording from an Intact Cockroach Nervous System

神经电路记录从一个完整的蟑螂神经系统

The cockroach ventral nerve cord preparation is a tractable system for neuroethology experiments, neural network modeling, and testing the physiological ...

Functional Analysis of the Larval Feeding Circuit in Drosophila

Functional Analysis of the Larval Feeding Circuit in Drosophila

在幼虫馈电电路的功能分析果蝇</i

The serotonergic  feeding circuit in Drosophila  melanogaster larvae can be used to investigate neuronal substrates of  critical importance during the ...

Time-dependent Increase in the Network Response to the Stimulation of Neuronal Cell Cultures on Micro-electrode Arrays

时间依赖性增加对微电极阵列刺激神经元细胞培养物的网络响应

Micro-electrode arrays (MEAs) can be used to investigate drug toxicity, design paradigms for next-generation personalized medicine, and study network ...