KPM支持欧盟FP7（RETICIRC）。

1。鱼的选育的胚胎被保留，并提出在标准条件下（品牌2002年）和天受精后（DPF）上演根据发展。成虫和幼虫在5 DPF用于测量。 2。实验过程<ol><li>制备仪器</li></ol>幼虫仔稚鱼被嵌入在3％预温（28°C），甲基纤维素，以防止身体动作。胚胎放在背侧在VisioTracker，面临的投影屏幕。成人鱼:鱼简要地在300毫克/升的MS-222麻醉，装入固定装置和对在VisioTracker下的。在开始测量之前，他们离开，恢复1-2分钟。 <ol start="2"><li>产生的刺激模式</li></ol>创建刺激模式由垂直的黑色和白色的正弦波光栅旋转周围的鱼使用专有的软件套件。它们可以是通过软件包调制根据波形，对比度，强度，角速度和空间频率。预计到使用数字光投影机在VisioTracker的，屏幕的模式。鱼的眼睛和屏幕之间的大致距离为4.5厘米，和投影尺寸屏幕上的水平方向360度和55度垂直。仔鱼，刺激方向与0.33赫兹的频率被改变，以减少扫视频率。成人鱼刺激，单向只眼睛的刺激颞鼻方向考虑，因为一般是鼻，颞眼速度显着降低和恒定的参考，以穆勒和Neuhauss的2010年。 <ol start="3"><li>记录眼球运动</li></ol>一个明场像鱼的头被送入红外线摄像机。红外照明鱼类的影响fROM如下。相机记录的影像在5帧/秒（幼虫）或12.5帧/秒（成人）的速度。图像自动处理，纠正眼睛的形状和平滑。眼睛有关的水平轴线上的方向，然后自动确定眼速度计算由专有软件套件。小运动的鱼被自动校正的软件。所有的记录和分析，实时完成。 3。实验后的数据处理<ol><li>眼睛的速度的信息的测量，过滤扫视，以便提取相缓慢的速度。 </li><li>扫视过滤眼速度曲线平滑的平均7帧的滑动窗口。 </li><li>眼睛的速度是相同的刺激条件下平均帧。 </li><li>仔鱼，双眼眼速度的平均值。 </li></ol> 4。代表性的成果: 对于仔鱼斑马鱼， 保险杠突变的选择。在此突变，晶状体上皮细胞hyperproliferate，导致减少的透镜尺寸和异位的镜头位置。这些形态学改变反映由对比敏感度的显着减少和视力（Schonthaler 等人，2010）。图1显示了差异对比敏感度的保险杠突变体与野生型的兄弟姐妹。 保险杠的突变越来越无法调整眼睛的刺激对比度下降的速度为。通过类推，当刺激空间频率的增加，即降低的刺激条纹宽度， 保险杠的突变体同样演示视力降低（图2） 成年斑马鱼visua的依赖升性能对环境条件的影响通过鱼经受不同的浓度的酒精，在他们的坦克水30分钟，并随后根据不同刺激条件下测量的视动响应。成人斑马鱼显示对比敏感度显着减少时，保持在体内酒精浓度增加（图3）。随着酒精浓度（图4）处理的鱼时，可观察到在很宽的范围内的空间频率类似的剂量依赖性降低眼睛的整体速度。此外，酒精治疗剂量依赖性地减少眼球运动性能更苛刻的任务，例如刺激的速度增加（图5）。 <img alt="图1" src="/files/ftp_upload/3556/3556fig1.jpg" /> 图1。眼睛斑马鱼幼虫的速度是依赖于刺激的对比。 10 保险杠突变体和野生型同胞10分析了在不同s 5的dpf<html> timulus条纹对比度的条件下。图表显示平均眼速度±1 SEM。 <img alt="图2" src="/files/ftp_upload/3556/3556fig2.jpg" /> 图2。斑马鱼幼体眼速度依赖于空间频率。进行10 保险杠突变体和野生型同胞10单丝旦数在5至不同的刺激条纹宽度，并分析所描述。图表显示平均眼速度±1 SEM。 <img alt="图3" src="/files/ftp_upload/3556/3556fig3.jpg" /> 图3。成人斑马鱼显示酒精浓度依赖性降低对比敏感度。成人斑马鱼保持在不同的酒精浓度为30分钟，并表示在不同的刺激条纹对比度条件分析。图表显示的平均时间鼻眼速度±1 SEM组（对照组除外:N = 11）9鱼。 E 4"SRC ="/ files/ftp_upload/3556/3556fig4.jpg"/&gt; 图4。成年斑马鱼秀酒精浓度依赖性降低整体的眼球运动的刺激条纹宽度在很宽的范围内。成人斑马鱼保持在不同的酒精浓度为30分钟，并表示在不同刺激的条的宽​​度条件分析。图表显示的平均时间鼻眼速度±1 SEM组（对照组除外:N = 11）9鱼。 <img alt="图5" src="/files/ftp_upload/3556/3556fig5.jpg" /> 图5。成人斑马鱼秀酒精的刺激速度的宽范围内的整体的眼球运动的浓度依赖性的降低。成人斑马鱼保持在不同的酒精浓度为30分钟，并表示根据不同刺激速度条件下进行分析。图表显示的平均时间鼻眼速度±1 SEM 9鱼组（对照组除外:N = 11）。

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奥利弗，DR Schnaedelbach和Holger D. Russig是TSE系统有限公司，生产员工在这篇文章中使用的可视化性能追踪系统。这篇文章TSE Systems GmbH公司主办的生产。斯蒂芬CF Neuhuass是苏黎世大学的雇员的接收报酬的TSE系统为每个系统出售。

OKR视觉功能的研究的重要性已在科学界公认的很长一段时间（复活节尼古拉·1996年，1997年），并在十年前已经开始尝试真正量化的典范。复活节和Nicola（1996）开发了一个系统，电动旋转​​条纹鼓，手动眼球运动的录像进行了分析。这个系统遭受从缺乏鱼胚胎，这需要频繁地手动重新定位的固定，并且可以只检测的眼睛跟踪运动以极大的困难。一个新的步骤是使用视频投影条纹鼓，让更多的变数计算机产生的刺激呈现（罗瑟和拜尔，2003年等Rinner。，2005年）。 大多是手动的，帧由帧录像录音的分析已被证明是非常费力的，并在一定程度上阻碍了观察者偏倚（Beck 等人。2004）。有人建议在实时的自动分析行为的反馈学习机制（Major 等，2004），以允许使用。使用红外照明和频率控制的旋转刺激已开创的Beck 等人 （2004）。然而，所描述的系统有只使用了幼虫，并进行了分析，脱机。此外，VisioTracker超过刺激，包括改变在实验过程中的刺激，从而允许更大的灵活性和自发的影响的实验过程中，允许完全控制。此外，用于创建数字激励由VisioTracker克服条纹刺激鼓（Beck 等人，2004年）的惰性质量的加速度与前面提到的问题。 幼虫的克制甲基不显着影响眼球运动，没有任何长期影响斑马鱼的福祉。仔稚鱼已成功保持在甲基纤维素嵌入了好几天，直到通过皮肤的氧气供应不足的需求随着年龄的增加（Qian 等，2005）。 成鱼的抑制方法是对动物也同样容易。的短持续时间的实验中，加上与迅速交换试验动物为一个不同的选项，还添加阳性动物福利方面的系统。由于鳃连续冲洗水，秒杀水和任何化学品的选择，以研究其对眼球运动和视觉表现很方便。同样，洗出实验可以被添加，而无需处理之间的实验动物。 的视频图像中的像素的噪声最小化通过平滑算法的专有VisioTracker软件，使眼睛的位置和角速度的高度精确的测量。此外，为方便统计分析，过滤软件扫视运动发生在固定的速度，不利于实验的说法。 7个视频帧的速度曲线的平均值为以后的分析提供了便利。 VisioTracker打开许多不同研究领域的一个新的维度。系统和它的前辈已经被成功地用于量化在斑马鱼幼虫的视觉效果，如视力，对比敏感度和光适应（Rinner 等人 ，2005年中，Schonthaler 等人，2010）使用参数，功能分析锥感光细胞处理视觉传导级联（例如Rinner 等，2005B，伦宁格等人 ，2011），或视觉缺陷的分析，突变的斑马鱼幼虫（例如Schonthaler 等，2005年，2008; Bahadori 等人，2006年）。成熟的视觉系统的形态和功能之间的相互依存关系已通过OKR测量研究，显示，视力为主，但不完全受限于感光间距在幼虫阶段（豪格等人，2010）。 VisioTracker是同样适用于分析在成年斑马鱼的视觉功能和其他类似的小型鱼类物种（穆勒和Neuhauss的（2010年），这份报告）。 还可以想到，从而以进行调查的物质可能被添加到周围的成年鱼鳃的水流的毒理学或药理学的研究领域中，如利用该系统。此外，通用性更彻底的分析的VisioTracker使例如，ontogenetics的视觉功能，神经回路的功能和发育，或感觉运动控制（见检讨黄Neuhauss，2008年）。

<table border="1"><tbody><tr><th>的试剂的名称</th><th>公司</th><th>目录编号</th></tr><tr><td>甲基纤维素</td><td> Sigma-Aldrich公司</td><td> M0387 </td></tr><tr><td> 3 - 氨基苯甲酸乙酯甲磺酸盐（MS-222） </td><td> Sigma-Aldrich公司</td><td> E10521 </td></tr><tr><td> 35毫米培养皿</td><td>康宁</td><td> 430165 </td></tr><tr><td>血清移液管</td><td>格雷纳生物一</td><td> 612 361 </td></tr><tr><td> VisioTracker </td><td> TSE系统</td><td> 302060 </td></tr></tbody></table>

visiotracker, an innovative automated approach to oculomotor analysis

视觉系统的发育和功能的调查必须量化行为模型的视觉效果，容易引起强大的，简单的操作。视动性反应（OKR），反身的行为，由于其高的选择价值存在于所有脊椎动物中已发现一个合适的模型。的的OKR涉及的与快速重扫视的眼睛交替缓慢的刺激变动如下。这种行为是很容易的测量在斑马鱼幼虫，由于其发病早期和稳定的（完全受精后96小时后（HPF）），以及受益于斑马鱼基因彻底的知识，几十年来的青睐模型在此输入的生物体。同时，在成鱼类似的机制分析得到了重视，尤其是对药理和毒理的应用程序。 这里我们描述VisioTracker的，完全自动化，高吞吐量T系统进行定量分析的视觉表现。该系统的基础上进行的研究组中的教授斯蒂芬Neuhauss的TSE系统被重新设计。它由一个固定装置配备一个高分辨率的变焦透镜由高品质的视频相机监视的小鱼。包围的鱼容器由一个鼓屏幕，计算机生成的刺激模式可以被投影。眼球运动的VisioTracker软件包，实时记录，并自动分析。 数据分析能够立即识别的参数，如慢速和快速阶段的持续时间，运动周期的频率，慢相增益，视力和对比敏感度。 典型的结果让例如药物或毒性和致突变的快速识别视觉系统的突变体，没有明显的改变，野生型形态，或确定的定量影响剂对视觉系统性能。

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VisioTracker, an Innovative Automated Approach to Oculomotor Analysis

在的VisioTracker是一个自动化的系统记录眼球运动的基础上，幼虫和小型成鱼的视觉性能的定量分析。它具有视觉刺激的属性和实时分析的完全控制权，使在视觉系统发育和功能，药理学，神经回路的研究和感觉运动整合等领域的高通量研究。

VisioTracker，创新的自动化到动眼神经分析方法

Investigations into the visual system development and function necessitate quantifiable behavioral models of visual performance that are easy to elicit, ...

visiotracker-an-innovative-automated-approach-to-oculomotor-analysis

Nanyang Technological University

Research

JoVE Journal

Neuroscience

11.0K 次观看.  University of Zurich. 在的VisioTracker是一个自动化的系统记录眼球运动的基础上，幼虫和小型成鱼的视觉性能的定量分析。它具有视觉刺激的属性和实时分析的完全控制权，使在视觉系统发育和功能，药理学，神经回路的研究和感觉运动整合等领域的高通量研究。

视频: VisioTracker, an Innovative Automated Approach to Oculomotor Analysis

Automated Sholl Analysis of Digitized Neuronal Morphology at Multiple Scales

Neuronal morphology plays a significant role in determining how neurons function and communicate1-3. Specifically, it affects the ability of neurons to receive inputs from other cells2 and contributes to the propagation of action potentials4,5. The morphology of the neurites also affects how information is processed. The diversity of dendrite morphologies facilitate local and long range signaling and allow individual neurons or groups of neurons to carry out specialized functions within the neuronal network6,7. Alterations in dendrite morphology, including fragmentation of dendrites and changes in branching patterns, have been observed in a number of disease states, including Alzheimer's disease8, schizophrenia9,10, and mental retardation11. The ability to both understand the factors that shape dendrite morphologies and to identify changes in dendrite morphologies is essential in the understanding of nervous system function and dysfunction.
Neurite morphology is often analyzed by Sholl analysis and by counting the number of neurites and the number of branch tips. This analysis is generally applied to dendrites, but it can also be applied to axons. Performing this analysis by hand is both time consuming and inevitably introduces variability due to experimenter bias and inconsistency. The Bonfire program is a semi-automated approach to the analysis of dendrite and axon morphology that builds upon available open-source morphological analysis tools. Our program enables the detection of local changes in dendrite and axon branching behaviors by performing Sholl analysis on subregions of the neuritic arbor. For example, Sholl analysis is performed on both the neuron as a whole as well as on each subset of processes (primary, secondary, terminal, root, etc.) Dendrite and axon patterning is influenced by a number of intracellular and extracellular factors, many acting locally. Thus, the resulting arbor morphology is a result of specific processes acting on specific neurites, making it necessary to perform morphological analysis on a smaller scale in order to observe these local variations12.
The Bonfire program requires the use of two open-source analysis tools, the NeuronJ plugin to ImageJ and NeuronStudio. Neurons are traced in ImageJ, and NeuronStudio is used to define the connectivity between neurites. Bonfire contains a number of custom scripts written in MATLAB (MathWorks) that are used to convert the data into the appropriate format for further analysis, check for user errors, and ultimately perform Sholl analysis. Finally, data are exported into Excel for statistical analysis. A flow chart of the Bonfire program is shown in Figure 1.

We have developed a computer program to analyze neuronal morphology. In combination with two existing open source analysis tools, our program performs Sholl analysis and determines the number of neurites, branch points, and neurite tips. The analyses are performed so that local changes in neurite morphology can be observed.

肖尔自动数字化的神经元形态分析，多尺度

Neuronal morphology plays a significant role in determining how neurons function and communicate1-3.  Specifically, it affects the ability of neurons to ...

科研

神经科学

VisualEyes: A Modular Software System for Oculomotor Experimentation

Eye movement studies have provided a strong foundation forming an understanding of how the brain acquires visual information in both the normal and dysfunctional brain.1 However, development of a platform to stimulate and store eye movements can require substantial programming, time and costs. Many systems do not offer the flexibility to program numerous stimuli for a variety of experimental needs. However, the VisualEyes System has a flexible architecture, allowing the operator to choose any background and foreground stimulus, program one or two screens for tandem or opposing eye movements and stimulate the left and right eye independently. This system can significantly reduce the programming development time needed to conduct an oculomotor study. The VisualEyes System will be discussed in three parts: 1) the oculomotor recording device to acquire eye movement responses, 2) the VisualEyes software written in LabView, to generate an array of stimuli and store responses as text files and 3) offline data analysis. Eye movements can be recorded by several types of instrumentation such as: a limbus tracking system, a sclera search coil, or a video image system. Typical eye movement stimuli such as saccadic steps, vergent ramps and vergent steps with the corresponding responses will be shown. In this video report, we demonstrate the flexibility of a system to create numerous visual stimuli and record eye movements that can be utilized by basic scientists and clinicians to study healthy as well as clinical populations.

Neural control and cognitive processes can be studied through eye movements. The VisualEyes software allows an operator to program stimuli on two computer screens independently using a simple, custom scripting language. The system can stimulate tandem eye movements (saccades and smooth pursuit) or opposing eye movements (vergence) or any combination.

VisualEyes:一个模块化的软件系统动眼神经实验

Eye movement studies have provided a strong foundation forming an understanding of how the brain acquires visual information in both the normal and ...

Brain Slice Biotinylation: An Ex Vivo Approach to Measure Region-specific Plasma Membrane Protein Trafficking in Adult Neurons

Regulated endocytic trafficking is the central mechanism facilitating a variety of neuromodulatory events, by dynamically controlling receptor, ion channel, and transporter cell surface presentation on a minutes time scale.&#160;There is a broad diversity of mechanisms that control endocytic trafficking of individual proteins. Studies investigating the molecular underpinnings of trafficking have primarily relied upon surface biotinylation to quantitatively measure changes in membrane protein surface expression in response to exogenous stimuli and gene manipulation.&#160;However, this approach has been mainly limited to cultured cells, which may not faithfully reflect the physiologically relevant mechanisms at play in adult neurons.&#160;Moreover, cultured cell approaches may underestimate region-specific differences in trafficking mechanisms.&#160;Here, we describe an approach that extends cell surface biotinylation to the acute brain slice preparation.&#160;We demonstrate that this method provides a high-fidelity approach to measure rapid changes in membrane protein surface levels in adult neurons.&#160;This approach is likely to have broad utility in the field of neuronal endocytic trafficking.

Brain Slice Biotinylation: An Ex Vivo Approach to Measure Region-specific Plasma Membrane Protein Trafficking in Adult Neurons

Neuronal membrane trafficking dynamically controls plasma membrane protein availability and significantly impacts neurotransmission.&#160;To date, it has been challenging to measure neuronal endocytic trafficking in adult neurons.&#160;Here, we describe a highly effective, quantitative method to measure rapid changes in surface protein expression ex vivo in acute brain slices.

大脑切片生物素化:一个<em&gt;前体内</em&gt;方法来衡量地区特有的质膜蛋白贩运成年神经元

Regulated endocytic trafficking is the central mechanism facilitating a variety of neuromodulatory events, by dynamically controlling receptor, ion ...

Interfacing 3D Engineered Neuronal Cultures to Micro-Electrode Arrays: An Innovative In Vitro Experimental Model

Currently, large-scale networks derived from dissociated neurons growing and developing in vitro on extracellular micro-transducer devices are the gold-standard experimental model to study basic neurophysiological mechanisms involved in the formation and maintenance of neuronal cell assemblies. However, in vitro studies have been limited to the recording of the electrophysiological activity generated by bi-dimensional (2D) neural networks. Nonetheless, given the intricate relationship between structure and dynamics, a significant improvement is necessary to investigate the formation and the developing dynamics of three-dimensional (3D) networks. In this work, a novel experimental platform in which 3D hippocampal or cortical networks are coupled to planar Micro-Electrode Arrays (MEAs) is presented. 3D networks are realized by seeding neurons in a scaffold constituted of glass microbeads (30-40 &#181;m in diameter) on which neurons are able to grow and form complex interconnected 3D assemblies. In this way, it is possible to design engineered 3D networks made up of 5-8 layers with an expected final cell density. The increasing complexity in the morphological organization of the 3D assembly induces an enhancement of the electrophysiological patterns displayed by this type of networks. Compared with the standard 2D networks, where highly stereotyped bursting activity emerges, the 3D structure alters the bursting activity in terms of duration and frequency, as well as it allows observation of more random spiking activity. In this sense, the developed 3D model more closely resembles in vivo neural networks.

Interfacing 3D Engineered Neuronal Cultures to Micro-Electrode Arrays: An Innovative In Vitro Experimental Model

In this work, a novel experimental model in which 3D neuronal cultures are coupled to planar Micro-Electrode Arrays (MEAs) is presented. 3D networks are built by seeding neurons in a scaffold made up of glass microbeads on which neurons grow and form interconnected 3D structures.

接口3D工程神经细胞微电极阵列:一种创新<em&gt;体外</em&gt;实验模型

Currently, large-scale networks derived from dissociated neurons growing and developing in vitro on extracellular micro-transducer devices are the ...

A Simple Alternative to Stereotactic Injection for Brain Specific Knockdown of miRNA

MicroRNAs (miRNAs) are key regulators of gene expression. In the brain, vital processes like neurodevelopment and neuronal functions depend on the correct expression of microRNAs. Perturbation of microRNAs in the brain can be used to model neurodegenerative diseases by modulating neuronal cell death. Currently, stereotactic injection is used to deliver miRNA knockdown agents to specific location in the brain. Here, we discuss strategies to design antagomirs against miRNA with locked nucleotide modifications (LNA). Subsequently describe a method for brain specific delivery of antagomirs, uniformly across different regions of the brain. This method is simple and widely applicable since it overcomes the surgery, associated injury and limitation of local delivery in stereotactic injections. We prepared a complex of neurotropic, cell-penetrating peptide Rabies Virus Glycoprotein (RVG) with antagomir against miRNA-29 and injected through tail vein, to specifically deliver in the brain. The antagomir design incorporated features that allow specific targeting of the miRNA and formation of non-covalent complexes with the peptide. The knock-down of the miRNA in neuronal cells, resulted in apoptotic cell death and associated behavioural defects. Thus, the method can be used for acute models of neuro-degeneration through the perturbation of miRNAs.

MicroRNAs play crucial roles in the brain and are potential targets for modeling neuro-degeneration. However, perturbing miRNA levels is challenging due to the short length of miRNA and inaccessibility of the brain tissue. This video presents a method for antagomir design and brain specific delivery using a neuropeptide in mice.

一个简单的替代，以立体定向注射miRNA的脑特异性敲除

MicroRNAs (miRNAs) are key regulators of gene expression.  In the brain, vital processes like neurodevelopment and neuronal functions depend on the ...

An Approach to Enhance Alignment and Myelination of Dorsal Root Ganglion Neurons

Axon regeneration is a chaotic process due largely to unorganized axon alignment. Therefore, in order for a sufficient number of regenerated axons to bridge the lesion site, properly organized axonal alignment is required. Since demyelination after nerve injury strongly impairs the conductive capacity of surviving axons, remyelination is critical for successful functioning of regenerated nerves. Previously, we demonstrated that mesenchymal stem cells (MSCs) aligned on a pre-stretch induced anisotropic surface because the cells can sense a larger effective stiffness in the stretched direction than in the perpendicular direction. We also showed that an anisotropic surface arising from a mechanical pre-stretched surface similarly affects alignment, as well as growth and myelination of axons. Here, we provide a detailed protocol for preparing a pre-stretched anisotropic surface, the isolation and culture of dorsal root ganglion (DRG) neurons on a pre-stretched surface, and show the myelination behavior of a co-culture of DRG neurons with Schwann cells (SCs) on a pre-stretched surface.

This protocol describes the isolation of dorsal root ganglion (DRG) neurons isolated from rats and the culture of DRG neurons on a static pre-stretched cell culture system to enhance axon alignment, with subsequent co-culture of Schwann Cells (SCs) to promote myelination.

一种方法来提高背根神经节的对齐和髓鞘

Axon regeneration is a chaotic process due largely to unorganized axon alignment. Therefore, in order for a sufficient number of regenerated axons to ...

Electrophysiological Method for Recording Intracellular Voltage Responses of Drosophila Photoreceptors and Interneurons to Light Stimuli In Vivo

Voltage responses of insect photoreceptors and visual interneurons can be accurately recorded with conventional sharp microelectrodes. The method described here enables the investigator to measure long-lasting (from minutes to hours) high-quality intracellular responses from single Drosophila R1-R6 photoreceptors and Large Monopolar Cells (LMCs) to light stimuli. Because the recording system has low noise, it can be used to study variability among individual cells in the fly eye, and how their outputs reflect the physical properties of the visual environment. We outline all key steps in performing this technique. The basic steps in constructing an appropriate electrophysiology set-up for recording, such as design and selection of the experimental equipment are described. We also explain how to prepare for recording by making appropriate (sharp) recording and (blunt) reference electrodes. Details are given on how to fix an intact fly in a bespoke fly-holder, prepare a small window in its eye and insert a recording electrode through this hole with minimal damage. We explain how to localize the center of a cell's receptive field, dark- or light-adapt the studied cell, and to record its voltage responses to dynamic light stimuli. Finally, we describe the criteria for stable normal recordings, show characteristic high-quality voltage responses of individual cells to different light stimuli, and briefly define how to quantify their signaling performance. Many aspects of the method are technically challenging and require practice and patience to master. But once learned and optimized for the investigator's experimental objectives, it grants outstanding in vivo neurophysiological data.

Electrophysiological Method for Recording Intracellular Voltage Responses of Drosophila Photoreceptors and Interneurons to Light Stimuli In Vivo

Sharp microelectrodes enable accurate electrophysiological characterization of photoreceptor and visual interneuron output in living Drosophila. Here we show how to use this method to record high-quality voltage responses of individual cells to controlled light stimulation. This method is ideal for studying neural information processing in insect compound eyes.

为记录细胞内电压响应电法<em&gt;果蝇</em&gt;光感受器和中间对光刺激<em&gt;在体内</em

Voltage responses of insect photoreceptors and visual interneurons can be accurately recorded with conventional sharp microelectrodes. The method ...

Automated Analysis of C. elegans Swim Behavior Using CeleST Software

Dissecting the neuronal and neuromuscular circuits that regulate behavior remains a major challenge in biology. The nematode Caenorhabditis elegans has proven to be an invaluable model organism in helping to tackle this challenge, from inspiring technological approaches,&#160;building the human brain connectome, to actually shedding light on the specific molecular drivers of basic functional patterns. The bulk of the behavioral studies in C. elegans have been performed on solid substrates. In liquid, animals exhibit behavioral patterns that include movement at a range of speeds in 3D, as well as partial body movements, such as a posterior curl without anterior shape change, which introduce new challenges for quantitation. The steps of a simple procedure, and use of a software that enables high-resolution analysis of C. elegans swim behavior, are presented here. The software, named CeleST, uses a specialized computer program that tracks multiple animals simultaneously and provides novel measures of C. elegans locomotion in liquid (swimming). The measures are mostly grounded in animal posture and based on mathematics used in computer vision and pattern recognition, without computational requirements for threshold cut-offs. The software tool can be used to both assess overall swimming prowess in hundreds of animals from combined small batch trials and to reveal novel phenotypes even in well-characterized genetic mutants. The preparation of specimens for analysis with CeleST is simple and low-tech, enabling wide adaptation by the scientific community. Use of the computational approach described here should therefore contribute to the greater understanding of behavior and behavioral circuits in the C. elegans model.

Automated Analysis of C. elegans Swim Behavior Using CeleST Software

An efficient and simple methodology for computer-based analysis of nematode swimming behavior in liquid is described. The method requires little to no investment for C. elegans laboratories. The hardware used is standard, and the computer software for the behavioral analysis (CeleST) is an open source one.

的自动分析<em&gt;℃。线虫</em&gt;游泳行为使用天书软件

Dissecting the neuronal and neuromuscular circuits that regulate behavior remains a major challenge in biology. The nematode Caenorhabditis elegans has ...

Automated Gait Analysis in Mice with Chronic Constriction Injury

The von Frey test is a classical method that has been widely used to examine the sensory function of neuropathic pain animals. However, it has some disadvantages such as subjective data and the requirement of a skilled, experienced experimenter. To date, a variety of modifications have improved the von Frey method, but it still has a few limitations. Recent reports have suggested that gait analysis produces more accurate and objective data from the neuropathic animals. This protocol demonstrates how to perform the automated gait analysis to determine the degree of neuropathic pain in mice. After several days of acclimation, the mice were allowed to walk freely on the glass floor to illuminate footprints. Then, quantification of the footprints and gait were performed through video clips with automatic analysis of various walking parameters, such as area of paw print, swing time, angle of paw, etc.
The main purpose of this study is to describe the methodology of automated gait analysis and briefly compare it with data from the classical sensory test using von Frey filament.

The precise assessment of pain response in a neuropathic animal model is critical to investigate the pathophysiology of pain diseases and develop new analgesics. We present a sensitive and objective method to determine the sensory function of the rodent hind paw by an automated gait analysis system.

慢性压迫性损伤小鼠步态的自动分析

The von Frey test is a classical method that has been widely used to examine the sensory function of neuropathic pain animals. However, it has some ...

Identification of Dopamine D1-Alpha Receptor Within Rodent Nucleus Accumbens by an Innovative RNA In Situ Detection Technology

In the central nervous system, the D1-alpha subtype receptor (Drd1&#945;) is the most abundant dopamine (DA) receptor, which plays a vital role in regulating neuronal growth and development. However, the mechanisms underlying Drd1&#945; receptor abnormalities mediating behavioral responses and modulating working memory function are still unclear. Using a novel RNA in situ hybridization assay, the current study identified dopamine Drd1&#945; receptor and tyrosine hydroxylase (TH) RNA expression from DA-related circuitry in the nucleus accumbens (NAc) area and substantia nigra region (SNR), respectively. Drd1&#945; expression in the NAc shows a &#34;discrete dot&#34; staining pattern. Clear sex differences in Drd1&#945; expression were observed. In contrast, TH shows a &#34;clustered&#34; staining pattern. Regarding TH expression, female rats displayed a higher signal expression per cell relative to male animals. The methods presented here provide a novel in situ hybridization technique for investigating changes in dopamine system dysfunction during the progression of central nervous system diseases.

Identification of Dopamine D1-Alpha Receptor Within Rodent Nucleus Accumbens by an Innovative RNA In Situ Detection Technology

Identification of dopamine D1-alpha receptor in the nucleus accumbens is critical for clarifying D1 receptor dysfunction during a central nervous system disease. We performed a novel RNA in situ hybridization assay to visualize single RNA molecules in a specific brain area.