作者要感谢全宗德拉RECHERCHE连接健康魁北克省（FRSQ），加拿大卫生研究院（CIHR），自然科学和工程研究理事会，加拿大（NSERC）的薪酬奖励，设备补助金，营运资金，和Concordia大学研究主席的计划（CRUC），以及在此稿件由简·斯图尔特博士颇有见地的反馈意见。

1。动物房<ol><li> 凯奇：为了记录的个人啮齿动物活动的运行轮，每个网箱应当拥有一个单一的啮齿动物和运行轮。由于正在运行的车轮可以被认为是一个形式丰富，在所有研究中的所有啮齿动物应该有类似的访问到正在运行的车轮。 </li><li> 床上用品的变化：动物的处理以及在笼子里或床上用品，都可以非光的昼夜节律的影响，21日至23日的 ，所以，笼网地板是理想的，因为他们尽量减少与动物的接触。尽管有这样一个托盘系统的可用性，床上用品应避免在关键阶段的实验。备选方案包括使用更持久的床上用品，这样可以让更多的罕见笼的变化，或不断变化的笼子放在一个伪随机的时间表。 </li><li> 隔离箱：网箱应保持在隔离箱，声音衰减，光合作ntrolled，通风良好。根据隔离框的大小和配置，在每个方块中的鸡笼的数目将通常范围从1-8。当住房在一个单一的隔离中的多个笼子里，人们应该知道，来自其他动物的各种气味和声音可以有昼夜节律的影响因子动物个体的行为。为了避免这些问题，应该尝试，以容纳一个笼子里每个隔离箱。 </li><li> 通风：足够的空气流动是必须的框一个舒适的家居环境为啮齿动物。在每个框中的风扇应该是罩住，以防止光到达的内部从外箱。此外，球迷们通常会去除空气从盒子里吹进了房间。小透光的通风口让空气得以进入隔离箱从几个点，并有助于避免不舒服的微风。为了验证是否有足够的通风，隔离箱内的温度（当封闭几个小时，用灯）应该是几乎相同的温度的房间里它被容纳。 </li><li> 照明：环境光的强度应该是相同的，在所有的笼子里。在类似的位置上面的每一个笼子里安排一个单独的光，并始终使用同一品牌/类型的灯泡。使用中等强度照明（100-300勒克斯）在笼级。避免过高的照明水平，这是更可能产生的昼夜系统， 本身 （ 例如掩模）的光，而不是直接修改应占行为。 </li><li> 黑暗/暗红色灯光：如果它是必要的处理或治疗动物在黑暗中（ 例如，在持续黑暗或夜间），应使用夜视镜。另外，由于昼夜节律系统是相对不敏感，红色波长，昏暗的红色灯光可以使用。您使用的具体红灯应进行测试，以确保它不会ALTER播放轮活动（ 如屏蔽），或调整生物钟（ 如产生相移）。 </li></ol> 2。数据收集（参见图1 - Vitalview硬件配置） <ol><li> 运行车轮的直径和人体工程学的行走轮将改变使用量24。因此，使用更小，更轻的轮毂和更大的小鼠，大鼠较重的车轮。当清洗并重新安装车轮，保证车轮​​可以旋转通畅，不要“摆动”，并记录微动开关被激活的车轮每转一圈的。 </li><li> 微动开关：传动轮的每个革命应该激活磁或机械微动开关。的微动开关的信息经由单个信道被发送，并记录由计算机可以纸槽随时间的数据（ 例如每2，5，6，或10分钟）。 </li><li> 计算机硬件：我们的伦尼NG轮的记录，则与Vitalview由微型发射机（ <a href="http://www.minimitter.com/vitalview_software.cfm" target="_blank">http://www.minimitter.com/vitalview_software.cfm</a> ）开发，硬件和软件平台。但是，也有其它记录平台如ClockLab，开发者Actimetrics（ <a href="http://www.actimetrics.com/ClockLab/" target="_blank">http://www.actimetrics.com/ClockLab/</a> ）。这两个平台汇集成一个单一的计算机文件数据从许多单个频道的信号源（ 例如，一个单一的微动开关由一个单一的运行轮激活）。从各个通道的数据，然后可以绘制和分析，分别在稍后的日期。 </li></ol> 3。数据记录<ol><li> 文件：上面提到的软件平台可用于分离出单一的渠道，使个别文件被创建为每个运行的轮记录。这些数据是最好的可视化和图形化特别DESIGNED程序，如Actiview（Minimitter，弯曲，OR），Circadia，或Clocklab的（Actimetrics，维尔梅特，IL），它可以生产周期图和actograms。然而，单声道文件也可以打开并使用一般的电子表格程序如Excel（微软，华盛顿州雷德蒙市）进行分析。 </li><li> 计算昼夜时间（CT）：CT 12，顾名思义，运行轮在夜间啮齿动物活动的发生。在24小时一天的同时，按照惯例，一昼夜被分解成24个昼夜小时。因此，如果自由运行的昼夜周期是24小时30分钟，所测得的壁时钟，CT 0将发生约12小时15分钟后，CT12。 </li></ol>

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没有利益冲突的声明。

监测日活动节律运行的车轮是最常用和最可靠的方法来评估在夜间啮齿类动物的主生物钟的输出。轮运行的活动，但是，只有一个，可以连续监测的行为和生理的许多方面。虽然绝大多数的磨合轮活动发生在夜间，超过30％的总觉醒发生在白天25,26。其他端点可用于评估，包括活动，食品斌方式，饮酒，睡眠，体温的昼夜节律。因此，根据性质的研究中，研究人员记录数的节奏同时进行。

<table border="1"&gt;<tbody&gt;<tr&gt;<td<strong&gt;的试剂名称</strong</td&gt;<td<strong&gt;公司</strong</td&gt;<td<strong&gt;产品目录号</strong</td&gt;<td<strong评论（可选）</strong</td&gt;</tr&gt;<tr&gt;<td&gt; Vitalview卡，配件及应用软件</td&gt;<td&gt;迷你发射器</td&gt;<td＃855-0030-00</td&gt;<td&gt;（弯管，OR，USA）</td&gt;</tr&gt;<tr&gt;<td&gt; DP24数据端口</td&gt;<td&gt;迷你发射器</td&gt;<td＃840-0024-00</td&gt;<td&gt;（弯管，OR，USA）</td&gt;</tr&gt;<tr&gt;<td&gt; QA4模块</td&gt;<td&gt;迷你发射器</td&gt;<td＃130-0050-00</td&gt;<td&gt;（弯管，OR，USA）</td&gt;</tr&gt;<tr&gt;<td&gt;磁性开关</td&gt;<td&gt;迷你发射器</td&gt;<td＃130-0015-00</td&gt;<td&gt;（弯管，OR，USA）</td&gt;</tr&gt;<tr&gt;<td&gt; C-50电缆组件</td&gt;<td&gt;迷你发射器</td&gt;<td＃060-0045-10</td&gt;<td&gt;（弯管，OR，USA）</td&gt;</tr&gt;<tr&gt;<td老鼠车轮总成</td&gt;<td&gt;迷你发射器</td&gt;<td＃640-0700-00</td&gt;<td&gt;（弯管，OR，USA）</td&gt;</tr&gt;<tr&gt;<td凯奇和托盘支持</td&gt;<td&gt;迷你发射器</td&gt;<td＃640-0400-00</td&gt;<td&gt;（弯管，OR，USA）</td&gt;</tr&gt;<tr&gt;<td&gt;可用切去笼</td&gt;<td&gt;迷你发射器</td&gt;<td＃664-2154-00</td&gt;<td&gt;（弯管，OR，USA）</td&gt;</tr&gt;<tr&gt;<td格地板笼</td&gt;<td&gt;迷你发射器</td&gt;<td＃676-2154-00</td&gt;<td&gt;（弯管，OR，USA）</td&gt;</tr&gt;<tr&gt;<td&gt;废盘</td&gt;<td&gt;迷你发射器</td&gt;<td＃684-2154-00</td&gt;<td&gt;（弯管，OR，USA）</td&gt;</tr&gt;<tr&gt;<td&gt;灯罩</td&gt;<td&gt;微晶科学</td&gt;<td＃R-101</td&gt;<td（多伦多，ON，加拿大）</td&gt;</tr&gt;<tr&gt;<td&gt; 4W荧光灯</td&gt;<td&gt;微晶科学</td&gt;<td＃F4T5/CW</td&gt;<td（多伦多，ON，加拿大）</td&gt;</tr&gt;<tr&gt;<td&gt;隔离室</td&gt;<td&gt;自定义建</td&gt;<td</td&gt;<td&gt; 28“H×20”W×28“D：½”黑三聚氰胺。</td&gt;</tr&gt;</tbody&gt;</table

recording and analysis of circadian rhythms in running-wheel activity in rodents

当啮齿动物可以免费使用在自己的笼子里一个跑轮，这轮自愿使用将取决于1-5天的时间。夜间啮齿动物，包括老鼠，仓鼠，老鼠，是活跃在夜间和白天的相对不活跃。许多其他的行为和生理的措施，也表现出的生活节奏，但在啮齿动物中，运行轮活动主生物钟，下丘脑的视交叉上核（SCN）的输出作为一个特别可靠和方便的措施。在一般情况下，通过一个过程被称为夹带，每天运行轮活动模式自然会与环境的明暗周期（LD周期， 例如 12小时光照：12小时暗）对齐。但昼夜节律是内源性产生的行为模式，表现出了〜24小时内，并坚持不断的黑暗。因此，在一个LD周期的情况下，记录和分析的行驶轮活动被用来确定主观时间 - 日期。因为这些导演的生物钟节律被称为主观时间天的昼夜时间（CT）。相反，当一个LD周期本，被称为天时间确定由环境LD周期给时的时间（ZT）。 虽然运行轮活动的昼夜节律，通常与SCN时钟6-8，也可能参与昼夜节律振荡器在许多其他地区的大脑和身体9-14日活动节律的调节。例如，生活节奏在食品预见性的活动不要求的SCN 15,16和取而代之的是，与额外的SCN振荡器17日至20日在活动中发生变化。因此，运行轮活动记录的行为提供了重要的信息不仅SCN的主时钟输出，同时也对额外的SCN振荡器的活动。下面我们DESCRIBE使用的设备和方法，记录，分析和显示啮齿类实验动物的自发活动节律昼夜。

<ol><li> 计算机程序：专门的计算机程序代的actograms和计算的昼夜周期中通常使用的。这些计划包括，但不限于，Actiview（弯，Minimitter，OR）和Circadia。 </li><li> actograms：Actograms提供每日的行驶轮活动模式的图解说明。有单密谋（x-轴= 24小时）和双绘制（x-轴= 48小时）actograms。这两种方法情节连续的天从上到下，但双绘制actograms情节两天就每一个水平线。具体而言，双，绘制actograms显示在最右...

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Recording and Analysis of Circadian Rhythms in Running-wheel Activity in Rodents

昼夜节律在自愿轮运行活动在哺乳动物中的紧密耦合的分子在大脑中的主时钟振荡。因此，这些生活节奏的行为可以被用来研究这个生物钟的功能，遗传，药物和环境因素的影响。

运行轮活动在啮齿类动物的昼夜节律的记录和分析

When rodents have free access to a running wheel in their home cage, voluntary use of this wheel will depend on the time of day1-5. Nocturnal rodents, ...

recording-analysis-circadian-rhythms-running-wheel-activity

Research

JoVE Journal

Neuroscience

21.1K 次观看.  McGill University. 昼夜节律在自愿轮运行活动在哺乳动物中的紧密耦合的分子在大脑中的主时钟振荡。因此，这些生活节奏的行为可以被用来研究这个生物钟的功能，遗传，药物和环境因素的影响。

视频: Recording and Analysis of Circadian Rhythms in Running-wheel Activity in Rodents

Assaying Locomotor Activity to Study Circadian Rhythms and Sleep Parameters in Drosophila

Most life forms exhibit daily rhythms in cellular, physiological and behavioral phenomena that are driven by endogenous circadian (&equiv;24 hr) pacemakers or clocks. Malfunctions in the human circadian system are associated with numerous diseases or disorders. Much progress towards our understanding of the mechanisms underlying circadian rhythms has emerged from genetic screens whereby an easily measured behavioral rhythm is used as a read-out of clock function. Studies using Drosophila have made seminal contributions to our understanding of the cellular and biochemical bases underlying circadian rhythms. The standard circadian behavioral read-out measured in Drosophila is locomotor activity. In general, the monitoring system involves specially designed devices that can measure the locomotor movement of Drosophila. These devices are housed in environmentally controlled incubators located in a darkroom and are based on using the interruption of a beam of infrared light to record the locomotor activity of individual flies contained inside small tubes. When measured over many days, Drosophila exhibit daily cycles of activity and inactivity, a behavioral rhythm that is governed by the animal's endogenous circadian system. The overall procedure has been simplified with the advent of commercially available locomotor activity monitoring devices and the development of software programs for data analysis. We use the system from Trikinetics Inc., which is the procedure described here and is currently the most popular system used worldwide. More recently, the same monitoring devices have been used to study sleep behavior in Drosophila. Because the daily wake-sleep cycles of many flies can be measured simultaneously and only 1 to 2 weeks worth of continuous locomotor activity data is usually sufficient, this system is ideal for large-scale screens to identify Drosophila manifesting altered circadian or sleep properties.

Assaying Locomotor Activity to Study Circadian Rhythms and Sleep Parameters in Drosophila

We describe procedures for recording daily locomotor activity rhythms of Drosophila and subsequent data analysis. Locomotor activity rhythms are a reliable behavioral output of animal circadian clocks and are used as the standard readout of clock function when studying circadian mutants or examining how the environment regulates the circadian system.

含量自发活动，研究昼夜节律和睡眠参数果蝇</em

Most life forms exhibit daily rhythms in cellular, physiological and behavioral phenomena that are driven by endogenous circadian (&equiv;24 hr) ...

科研

神经科学

Measuring Circadian and Acute Light Responses in Mice using Wheel Running Activity

Circadian rhythms are physiological functions that cycle over a period of approximately 24 hours (circadian- circa: approximate and diem: day)1, 2. They are responsible for timing our sleep/wake cycles and hormone secretion. Since this timing is not precisely 24-hours, it is synchronized to the solar day by light input. This is accomplished via photic input from the retina to the suprachiasmatic nucleus (SCN) which serves as the master pacemaker synchronizing peripheral clocks in other regions of the brain and peripheral tissues to the environmental light dark cycle3-7. The alignment of rhythms to this environmental light dark cycle organizes particular physiological events to the correct temporal niche, which is crucial for survival8. For example, mice sleep during the day and are active at night. This ability to consolidate activity to either the light or dark portion of the day is referred to as circadian photoentrainment and requires light input to the circadian clock9. Activity of mice at night is robust particularly in the presence of a running wheel. Measuring this behavior is a minimally invasive method that can be used to evaluate the functionality of the circadian system as well as light input to this system. Methods that will covered here are used to examine the circadian clock, light input to this system, as well as the direct influence of light on wheel running behavior.

This article will review methods that can be used to determine circadian function and light responsiveness in mice.

测量小鼠使用车轮运行活动的昼夜和急性对光反应

Circadian rhythms are physiological functions that cycle over a period of approximately 24 hours (circadian- circa: approximate and diem: day)1, 2.  They ...

Large-scale Recording of Neurons by Movable Silicon Probes in Behaving Rodents

A major challenge in neuroscience is linking behavior to the collective activity of neural assemblies. Understanding of input-output relationships of neurons and circuits requires methods with the spatial selectivity and temporal resolution appropriate for mechanistic analysis of neural ensembles in the behaving animal, i.e. recording of representatively large samples of isolated single neurons. Ensemble monitoring of neuronal activity has progressed remarkably in the past decade in both small and large-brained animals, including human subjects1-11. Multiple-site recording with silicon-based devices are particularly effective because of their scalability, small volume and geometric design.
Here, we describe methods for recording multiple single neurons and local field potential in behaving rodents, using commercially available micro-machined silicon probes with custom-made accessory components. There are two basic options for interfacing silicon probes to preamplifiers: printed circuit boards and flexible cables. Probe supplying companies (<a href="http://www.neuronexustech.com/" >http://www.neuronexustech.com/</a>; <a href="http://www.sbmicrosystems.com/">http://www.sbmicrosystems.com/</a>; <a href="http://www.acreo.se/">http://www.acreo.se/</a>) usually provide the bonding service and deliver probes bonded to printed circuit boards or flexible cables. Here, we describe the implantation of a 4-shank, 32-site probe attached to flexible polyimide cable, and mounted on a movable microdrive. Each step of the probe preparation, microdrive construction and surgery is illustrated so that the end user can easily replicate the process.

We describe methods for large-scale recording of multiple single units and local field potential in behaving rodents with silicon probes. Drive fabrication, probe attachment to the drive and probe implantation processes are illustrated in sufficient details for easy replication.

移动硅探针的神经行为鼠害的大型记录

A major challenge in neuroscience is linking behavior to the collective activity of neural assemblies. Understanding of input-output relationships of ...

A Fully Automated and Highly Versatile System for Testing Multi-cognitive Functions and Recording Neuronal Activities in Rodents

We have developed a fully automated system for operant behavior testing and neuronal activity recording by which multiple cognitive brain functions can be investigated in a single task sequence. The unique feature of this system is a custom-made, acoustically transparent chamber that eliminates many of the issues associated with auditory cue control in most commercially available chambers. The ease with which operant devices can be added or replaced makes this system quite versatile, allowing for the implementation of a variety of auditory, visual, and olfactory behavioral tasks. Automation of the system allows fine temporal (10 ms) control and precise time-stamping of each event in a predesigned behavioral sequence. When combined with a multi-channel electrophysiology recording system, multiple cognitive brain functions, such as motivation, attention, decision-making, patience, and rewards, can be examined sequentially or independently.

In this report, we present a fully automated and highly versatile system capable of simultaneously testing multi-cognitive behaviors and recording neuronal activities for rodents.

一个多认知功能测试，并记录在啮齿动物的神经元活动的完全自动化和高度灵活的系统

We have developed a fully automated system for operant behavior testing and neuronal activity recording by which multiple cognitive brain functions can be ...

Retrograde Fluorescent Labeling Allows for Targeted Extracellular Single-unit Recording from Identified Neurons In vivo

The overall goal of this method is to record single-unit responses from an identified population of neurons. In vivo electrophysiological recordings from individual neurons are critical for understanding how neural circuits function under natural conditions. Traditionally, these recordings have been performed 'blind', meaning the identity of the recorded cell is unknown at the start of the recording. Cellular identity can be subsequently determined via intracellular1, juxtacellular2 or loose-patch3 iontophoresis of dye, but these recordings cannot be pre-targeted to specific neurons in regions with functionally heterogeneous cell types. Fluorescent proteins can be expressed in a cell-type specific manner permitting visually-guided single-cell electrophysiology4-6. However, there are many model systems for which these genetic tools are not available. Even in genetically accessible model systems, the desired promoter may be unknown or genetically homogenous neurons may have varying projection patterns. Similarly, viral vectors have been used to label specific subgroups of projection neurons7, but use of this method is limited by toxicity and lack of trans-synaptic specificity. Thus, additional techniques that offer specific pre-visualization to record from identified single neurons in vivo are needed. Pre-visualization of the target neuron is particularly useful for challenging recording conditions, for which classical single-cell recordings are often prohibitively difficult8-11. The novel technique described in this paper uses retrograde transport of a fluorescent dye applied using tungsten needles to rapidly and selectively label a specific subset of cells within a particular brain region based on their unique axonal projections, thereby providing a visual cue to obtain targeted electrophysiological recordings from identified neurons in an intact circuit within a vertebrate CNS.
The most significant novel advancement of our method is the use of fluorescent labeling to target specific cell types in a non-genetically accessible model system. Weakly electric fish are an excellent model system for studying neural circuits in awake, behaving animals12. We utilized this technique to study sensory processing by "small cells" in the anterior exterolateral nucleus (ELa) of weakly electric mormyrid fish. "Small cells" are hypothesized to be time comparator neurons important for detecting submillisecond differences in the arrival times of presynaptic spikes13. However, anatomical features such as dense myelin, engulfing synapses, and small cell bodies have made it extremely difficult to record from these cells using traditional methods11, 14. Here we demonstrate that our novel method selectively labels these cells in 28% of preparations, allowing for reliable, robust recordings and characterization of responses to electrosensory stimulation.

Retrograde Fluorescent Labeling Allows for Targeted Extracellular Single-unit Recording from Identified Neurons In vivo

Retrograde transport of fluorescent dye labels a sub-population of neurons based on anatomical projection. Labeled axons can be visually targeted in vivo, permitting extracellular recording from identified axons. This technique facilitates recording when neurons cannot be labeled through genetic manipulation or are difficult to isolate using 'blind' in vivo approaches.

逆行荧光染料标记允许有针对性的外单细胞记录技术鉴定神经元<em&gt;在体内</em

The overall goal of this method is to record single-unit responses from an identified population of neurons. In vivo electrophysiological recordings from ...

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

Detection and interpretation of olfactory cues are critical for the survival of many organisms. Remarkably, species across phyla have strikingly similar olfactory systems suggesting that the biological approach to chemical sensing has been optimized over evolutionary time1. In the insect olfactory system, odorants are transduced by olfactory receptor neurons (ORN) in the antenna, which convert chemical stimuli into trains of action potentials. Sensory input from the ORNs is then relayed to the antennal lobe (AL; a structure analogous to the vertebrate olfactory bulb). In the AL, neural representations for odors take the form of spatiotemporal firing patterns distributed across ensembles of principal neurons (PNs; also referred to as projection neurons)2,3. The AL output is subsequently processed by Kenyon cells (KCs) in the downstream mushroom body (MB), a structure associated with olfactory memory and learning4,5. Here, we present electrophysiological recording techniques to monitor odor-evoked neural responses in these olfactory circuits.
First, we present a single sensillum recording method to study odor-evoked responses at the level of populations of ORNs6,7. We discuss the use of saline filled sharpened glass pipettes as electrodes to extracellularly monitor ORN responses. Next, we present a method to extracellularly monitor PN responses using a commercial 16-channel electrode3. A similar approach using a custom-made 8-channel twisted wire tetrode is demonstrated for Kenyon cell recordings8. We provide details of our experimental setup and present representative recording traces for each of these techniques.

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

We demonstrate variations of the extracellular multi-unit recording technique to characterize odor-evoked responses in the first three stages of the invertebrate olfactory pathway. These techniques can easily be adapted to examine ensemble activity in other neural systems as well.

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 ...

Deep Brain Stimulation with Simultaneous fMRI in Rodents

In order to visualize the global and downstream neuronal responses to deep brain stimulation (DBS) at various targets, we have developed a protocol for using blood oxygen level dependent (BOLD) functional magnetic resonance imaging (fMRI) to image rodents with simultaneous DBS. DBS fMRI presents a number of technical challenges, including accuracy of electrode implantation, MR artifacts created by the electrode, choice of anesthesia and paralytic to minimize any neuronal effects while simultaneously eliminating animal motion, and maintenance of physiological parameters, deviation from which can confound the BOLD signal.&#160;Our laboratory has developed a set of procedures that are capable of overcoming most of these possible issues. For electrical stimulation, a homemade tungsten bipolar microelectrode is used, inserted stereotactically at the stimulation site in the anesthetized subject. In preparation for imaging, rodents are fixed on a plastic headpiece and transferred to the magnet bore. For sedation and paralysis during scanning, a cocktail of dexmedetomidine and pancuronium is continuously infused, along with a minimal dose of isoflurane; this preparation minimizes the BOLD ceiling effect of volatile anesthetics. In this example experiment, stimulation of the subthalamic nucleus (STN) produces BOLD responses which are observed primarily in ipsilateral cortical regions, centered in motor cortex. Simultaneous DBS and fMRI allows the unambiguous modulation of neural circuits dependent on stimulation location and stimulation parameters, and permits observation of neuronal modulations free of regional bias. This technique may be used to explore the downstream effects of modulating neural circuitry at nearly any brain region, with implications for both experimental and clinical DBS.

This protocol describes a standard method for simultaneous functional magnetic resonance imaging and deep brain stimulation in the rodent. The combined use of these experimental tools allows for the exploration of global downstream activity in response to electrical stimulation at virtually any brain target.

深部脑刺激与同步功能磁共振成像在鼠害

In order to visualize the global and downstream neuronal responses to deep brain stimulation (DBS) at various targets, we have developed a protocol for ...

In Vitro Recording of Mesenteric Afferent Nerve Activity in Mouse Jejunal and Colonic Segments

Afferent nerves not only convey information concerning normal physiology, but also signal disturbed homeostasis and pathophysiological processes of the different organ systems from the periphery towards the central nervous system. As such, the increased activity or &#39;sensitization&#39; of mesenteric afferent nerves has been allocated an important role in the pathophysiology of visceral hypersensitivity and abdominal pain syndromes.
Mesenteric afferent nerve activity can be measured in vitro in an isolated intestinal segment that is mounted in a purpose-built organ bath and from which the splanchnic nerve is isolated, allowing researchers to directly assess nerve activity adjacent to the gastrointestinal segment. Activity can be recorded at baseline in standardized conditions, during distension of the segment or following the addition of pharmacological compounds delivered intraluminally or serosally. This technique allows the researcher to easily study the effect of drugs targeting the peripheral nervous system in control specimens; besides, it provides crucial information on how neuronal activity is altered during disease. It should be noted however that measuring afferent neuronal firing activity only constitutes one relay station in the complex neuronal signaling cascade, and researchers should bear in mind not to overlook neuronal activity at other levels (e.g., dorsal root ganglia, spinal cord or central nervous system) in order to fully elucidate the complex neuronal physiology in health and disease.
Commonly used applications include the study of neuronal activity in response to the administration of lipopolysaccharide, and the study of afferent nerve activity in animal models of irritable bowel syndrome. In a more translational approach, the isolated mouse intestinal segment can be exposed to colonic supernatants from IBS patients. Furthermore, a modification of this technique has been recently shown to be applicable in human colonic specimens.

In Vitro Recording of Mesenteric Afferent Nerve Activity in Mouse Jejunal and Colonic Segments

Mesenteric afferent nerves convey information from the gastrointestinal tract towards the brain regarding normal homeostasis as well as pathophysiology. Gastrointestinal afferent nerve activity can be assessed by mounting isolated intestinal segments with attached afferent nerves into an organ bath, isolating the nerve, and assessing basal as well as stimulated activity.

在鼠标空肠和结肠段肠系膜传入神经活性的体外记录

Afferent nerves not only convey information concerning normal physiology, but also signal disturbed homeostasis and pathophysiological processes of the ...

Preparations and Protocols for Whole Cell Patch Clamp Recording of Xenopus laevis Tectal Neurons

The Xenopus tadpole retinotectal circuit, comprised of the retinal ganglion cells (RGCs) in the eye which form synapses directly onto neurons in the optic tectum, is a popular model to study how neural circuits self-assemble. The ability to carry out whole cell patch clamp recordings from tectal neurons and to record RGC-evoked responses, either in vivo or using a whole brain preparation, has generated a large body of high-resolution data about the mechanisms underlying normal, and abnormal, circuit formation and function. Here we describe how to perform the in vivo preparation, the original whole brain preparation, and a more recently developed horizontal brain slice preparation for obtaining whole cell patch clamp recordings from tectal neurons. Each preparation has unique experimental advantages. The in vivo preparation enables the recording of the direct response of tectal neurons to visual stimuli projected onto the eye. The whole brain preparation allows for the RGC axons to be activated in a highly controlled manner, and the horizontal brain slice preparation allows recording from across all layers of the tectum.

Preparations and Protocols for Whole Cell Patch Clamp Recording of Xenopus laevis Tectal Neurons

In this paper, we discuss three brain preparations used for whole cell patch clamp recording to study the retinotectal circuit of Xenopus laevis tadpoles. Each preparation, with its own specific advantages, contributes to the experimental tractability of the Xenopus tadpole as a model to study neural circuit function.

爪蟾Tectal 神经元全细胞膜片钳记录的制备与协议

The Xenopus tadpole retinotectal circuit, comprised of the retinal ganglion cells (RGCs) in the eye which form synapses directly onto neurons in the optic ...

The Power of Interstimulus Interval for the Assessment of Temporal Processing in Rodents

Temporal processing deficits have been implicated as a potential elemental dimension of higher-level cognitive processes, commonly observed in neurocognitive disorders. Despite the popularization of prepulse inhibition (PPI) in recent years, many current protocols promote using a percent of control measure, thereby precluding the assessment of temporal processing. The present study used cross-modal PPI and gap prepulse inhibition (gap-PPI) to demonstrate the benefits of employing a range of interstimulus intervals (ISIs) to delineate effects of sensory modality, psychostimulant exposure, and age. Assessment of sensory modality, psychostimulant exposure, and age reveals the utility of an approach varying the interstimulus interval (ISI) to establish the shape of the ISI function, including increases (sharper curve inflections) or decreases (flattening of the response amplitude curve) in startle amplitude. Additionally, shifts in peak response inhibition, suggestive of a differential sensitivity to the manipulation of ISI, are often revealed. Thus, the systematic manipulation of ISI affords a critical opportunity to evaluate temporal processing, which may reveal the underlying neural mechanisms involved in neurocognitive disorders.

Temporal processing, a preattentive process, may underlie deficits in higher-level cognitive processes, including attention, commonly observed in neurocognitive disorders. Using prepulse inhibition as an exemplar paradigm, we present a protocol for manipulating interstimulus interval (ISI) to establish the shape of the ISI function to provide an assessment of temporal processing.