这项工作是由威康信托基金资助。

任何研究之前，可进行婴儿“的伦理审批和知情的书面需要寻求家长的同意。 获得大学医院伦理委员会在这项研究中的伦理批准，并告知父母的书面同意之前，每个过程中获得。这项研究符合赫尔辛基宣言所定的标准和良好的临床实践指南。 1。数据收集 - 设置<ol><li>至少16个人可支配的脑电图Ag / AgCl电极杯电极放在婴儿的头部皮肤后预备10-20电极放置系统（图1A）。可以实现更全面的覆盖，使用与嵌入电极的脑电图第。使用脑电图帽的过程更快，破坏性较小，尤其是作为电极与感性凝胶放置在头上的帽前的准备。一次性脑电图电极的应用要求更多的时间和技能，但一般在一个更好的录音结果。考虑减少电极的数量是有限的，如果访问的婴儿，但总是用中线电极（CZ，CPZ和FCZ）。 FCZ使用录音作为参考电极。 </li><li>使用脑电图导电膏优化电极/皮肤电耦合。 </li><li>放置在电极的弹性网举行的地方。 </li><li>铁电极的信息一起，以尽量减少电气干扰。 </li><li>到胸部或头部放置一个接地电极。 </li><li>广场上的两条腿的股二头肌肌电电极后皮肤预备（ 图1B） 。 </li><li>要记录到人体的心电活动场所导致1心电电极后皮肤预备（胸部左侧的一个电极，一个正确的，并使用相同的接地电极脑电图）。 </li><li>放置在腹部来衡量呼吸的运动传感器。 </li><li>脉搏血氧饱和度探头放置在脚下测量血氧饱和度和心率。确保探头是否到位和连续信号记录无辍学担保。血氧饱和度探头必须是对侧脚，你打算以刺激的脚放在。 </li><li>三脚架安装摄像机设置帧的婴儿的脸，使面部表情的变化可以记录。 </li><li>放置在相机内的发光二极管（LED）。 LED是联系在一起的时序电路，所以它会闪光刺激时同步脑电图，肌电图和录像。 </li></ol> 2。数据收集 - 记录<ol><li>开始录像。 </li><li>开始录制脉搏血氧饱和度。 </li><li>开始脑电/肌电图记录。 </li><li>等待，直到婴儿落户。 </li><li>按住脚，如果执行脚跟枪和手动事件标记的EEG和EMG录音。这个时代将被用来确定一个控件的背景部分脑电图肌电图。 </li><li>应用对表面脚跟肌腱锤臂是连接到一个橡胶塞轻轻拍打触摸刺激。刺激，不重视脚的脉搏血氧饱和度。当婴儿刺激的脑电/肌电图和录像，必须事件标记，以确定在录音时，当刺激发生。触摸刺激可事件标记附加阻抗头肌腱锤电子链接的刺激的录音设备。视频录制事件标志着LED闪光灯。反复接触，可应用于可以应用到身体的不同地区和刺激，即肩</li><li>应用旋转90度的柳叶刀，并把对脚，所以弹簧加载刀片被释放时，它不接触皮肤有害的非控制刺激。此事件可使用附加顶面的T加速度时间锁定他枪。加速度计检测刀片被释放时发生振动。 </li><li>按照在新生儿病房的临床实践中执行临床必需的鞋跟枪。等待，直到脑电活动是做脚跟枪之前解决。脚跟枪的时间锁定可以以同样的方式进行控制的刺激。脚跟枪之后，不挤至少30秒的脚，以确保记录的反应完全是由于兰斯。 </li><li>确保所需的血液量是收集和准备的临床分析样本。 </li><li>保存数据，并停止所有的录音。 </li><li>记录婴儿的人口统计资料和实验的细节，并输入到数据库安全储存和日后参考。 </li><li>重复此过程中的婴儿所需的样本。在这个例子中的婴儿数= 23。 </li></ol> 3。脑电数据分析<ol><li>创建1.7秒对应于每个接触，控制和长矛刺激和背景脑电图脑电图时代。时代应该开始每个事件前0.6秒。每个模式对应的时代，应该是相同的。 </li><li>基线正确减去平均基线信号和高通滤波器在0.1赫兹的时代。 </li><li>考虑CPZ或Cz的记录的时代，作进一步的分析和排除这是由运动的人为污染的时代。运动的人为被定义为一个小于50ms的变化幅度超过50μV的。 </li><li>对齐的痕迹纠正延迟50至300毫秒后的刺激，并进行主成分分析（PCA）在此时间间隔抖动识别触觉的潜力（即脑电活动有关的触觉刺激）。考虑的时代，是变量和时间点的意见。 PCA分解成基本波形脑电图的时代，称为主成分（PC）和代表在跨越时间点的信号的振幅系统的变化。 </li><li>运行分析单向方差（ANOVA）的第2个，每年的重量，以确定哪台PC触觉潜力。这将是个人电脑，其重量均显著较大以下触觉刺激的背景脑电图相比。 </li><li>调整的痕迹，以正确的延迟抖动之间的300和700毫秒后的刺激，并进行在此时间间隔的PCA。 </li><li>运行单向方差分析的第2个，每年的重量，以确定哪台PC伤害性的具体潜力。这将是个人电脑，其重量均显著较大的下列有害刺激，触觉刺激和背景脑电图相比。 </li></ol> 4。肌电图的数据分析<ol><li>计算根均方根（RMS）在1000毫秒后刺激的控制和长矛刺激的肌电信号。 </li><li>执行T -测试RMS值，以确定具体的伤害性脊髓反射撤出。 </li></ol> 5。代表性的成果<img alt="图1" src="/files/ftp_upload/3118/3118fig1.jpg" /> <st荣&gt;图1（A）（I）图及（ii）（修改国际10/20电极安置系统）的脑电图电极配售的照片;（B）图肌电录音电极安置股二头肌。 <img alt="图2" src="/files/ftp_upload/3118/3118fig2.jpg" /> 。图2（a）在Cz感官潜力的例子诱发接触3名婴儿;（二）由3名婴儿的有害枪伤害性的具体潜力的例子，在Cz诱发。 <img alt="图3" src="/files/ftp_upload/3118/3118fig3.jpg" /> 图3。刺激方式在Cz PC重量（平均± SEM）的依赖。 50-300毫秒之间获得刺激后发病的PC代表了触觉的潜力和300-700毫秒之间获得刺激后发病的PC代表一个特定的伤害性的潜力。覆盖在盛大的平均电脑（粗线）在所有的刺激类型（背景脑电图，触摸，有毒枪）获得个人的痕迹后在规定的时间间隔保持一致。 <img alt="图4" src="/files/ftp_upload/3118/3118fig4.jpg" /> 图4：（一）后肌电活动中的一个婴儿的范例（一）有毒脚跟枪及（ii）触摸脚跟。 （二）平均（± SE）肌电图根均方根肌电图后鞋跟有毒，有害的非触摸刺激婴儿的活动。

<ol>
	<li>Fitzgerald, M., Walker, S. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Infant+pain+management:+a+developmental+neurobiological+approach.">Infant pain management: a developmental neurobiological approach.</a> Nat. Clin. Pract. Neurol. 5 (1), 35-35 (2009).</li><li>Slater, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Oral+sucrose+as+an+analgesic+drug+for+procedural+pain+in+newborn+infants:+a+randomised+controlled+trial.">Oral sucrose as an analgesic drug for procedural pain in newborn infants: a randomised controlled trial.</a> Lancet. 376 (9748), 1225-1225 (2010).</li><li>Slater, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Evoked+potentials+generated+by+noxious+stimulation+in+the+human+infant+brain.">Evoked potentials generated by noxious stimulation in the human infant brain.</a> Eur. J. Pain. 14 (3), 321-321 (2010).</li><li>Slater, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Premature+infants+display+increased+noxious-evoked+neuronal+activity+in+the+brain+compared+to+healthy+age-matched+term-born+infants.">Premature infants display increased noxious-evoked neuronal activity in the brain compared to healthy age-matched term-born infants.</a> Neuroimage. 52 (2), 583-583 (2010).</li></ol>

作者宣称，他们什么都没有透露。

该视频演示如何通过触觉和伤害性刺激诱发，电生理反应，可以使用EEG和EMG记录在人类婴儿的特点。这种类型的研究，将有助于了解人类的疼痛处理的发展和可塑性，并导致改善临床评估和治疗婴幼儿疼痛1， 2。 这些实验的成功，需要一个多学科小组的密切合作。需要在新生儿科，临床神经生理学，生物工程和发育神经科学领域的专业知识。 重要的是，婴儿的护理最高优先级，当这些实验进行。研究护士或医生执行脚跟枪婴儿的福祉的责任和需要，以确保实验是根据临床实践。从医院伦理委员会和写法的批准ñ父母同意有必要进行这些研究。 脚跟枪是临床需要，并不能反复研究的目的。因此，必须是在一个安全可靠，功能强大的方式，不干预与临床实践3，4的收集，录音。特别是，它是至关重要的，时间锁用于刺激的方法是可靠的，并不妨碍临床采血。 所有的婴儿疼痛的措施必然是间接的，因此它是重要的，以确定记录的活动是专门的有毒事件，并没有其他方面的照顾婴儿的2诱发。通过仔细的技术设计和数据处理后，要做到这一点。

electrophysiological measurements and analysis of nociception in human infants

疼痛是一种不愉快的感觉和情绪​​体验。由于婴儿不能口头报告他们的经验，目前疼痛评估方法是基于行为和生理的身体反应，如哭闹，身体动作或面部表情的变化，。虽然这些措施表明，婴儿装入一个伤害性刺激后的反应，他们是有限的：他们是基于激活皮层躯体和植物神经电机的途径，可能无法可靠地连接到中央在大脑的感官加工。中枢神经系统如何响应有毒事件的知识，可以提供一个伤害性信息和疼痛是如何处理新生儿的洞察力。 足跟采血程序，用于提取住院婴儿的血液，提供了一个独特的机会来研究在婴儿期的痛苦。在这个视频中，我们描述了脑电图（EEG）和肌电图（EMG）的时间锁定此过程CAn是用​​于研究大脑和脊髓伤害性活动。 这种综合的方法来测量婴儿疼痛，有潜力，一个有效的和敏感的临床测量工具铺平方式。

Watch this Scientific Journal Video about Electrophysiological Measurements and Analysis of Nociception in Human Infants at JoVE.com

Electrophysiological Measurements and Analysis of Nociception in Human Infants

婴幼儿疼痛评估和治疗是困难的，因为婴儿不能口头报告他们的经验。在这个视频中，我们描述了定量的电生理方法和分析技术，可用于测量从婴儿神经系统的有毒事件的反应。

在人类婴儿的伤害性电生理测量和分析

Pain is an unpleasant sensory and emotional experience. Since infants cannot verbally report their experiences, current methods of pain assessment are ...

electrophysiological-measurements-analysis-nociception-human

Research

JoVE Journal

Neuroscience

17.1K 次观看.  University College London. 婴幼儿疼痛评估和治疗是困难的，因为婴儿不能口头报告他们的经验。在这个视频中，我们描述了定量的电生理方法和分析技术，可用于测量从婴儿神经系统的有毒事件的反应。

视频: Electrophysiological Measurements and Analysis of Nociception in Human Infants

Electrophysiological Measurements from a Moth Olfactory System

Insect olfactory systems provide unique opportunities for recording odorant-induced responses in the forms of electroantennograms (EAG) and single sensillum recordings (SSR), which are summed responses from all odorant receptor neurons (ORNs) located on the antenna and from those housed in individual sensilla, respectively. These approaches have been exploited for getting a better understanding of insect chemical communication. The identified stimuli can then be used as either attractants or repellents in management strategies for insect pests.

Insect olfactory systems provide unique opportunities for recording odorant-induced responses in the forms of electroantennograms (EAG) and single sensillum recordings (SSR), which are summed responses from all odorant receptor neurons (ORNs) located on the antenna and from those housed in individual sensilla, respectively.

从蛾的嗅觉系统的电生理测量

Insect olfactory systems provide unique opportunities for recording odorant-induced responses in the forms of electroantennograms (EAG) and single ...

科研

神经科学

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

Precise patterns of division, migration and differentiation of neural progenitor cells are crucial for proper brain development and function1,2. To understand the behavior of neural progenitor cells in the complex in vivo environment, time-lapse live imaging of neural progenitor cells in an intact brain is critically required. In this video, we exploit the unique features of zebrafish embryos to visualize the development of forebrain neural progenitor cells in vivo. We use electroporation to genetically and sparsely label individual neural progenitor cells. Briefly, DNA constructs coding for fluorescent markers were injected into the forebrain ventricle of 22 hours post fertilization (hpf) zebrafish embryos and electric pulses were delivered immediately. Six hours later, the electroporated zebrafish embryos were mounted with low melting point agarose in glass bottom culture dishes. Fluorescently labeled neural progenitor cells were then imaged for 36hours with fixed intervals under a confocal microscope using water dipping objective lens. The present method provides a way to gain insights into the in vivo development of forebrain neural progenitor cells and can be applied to other parts of the central nervous system of the zebrafish embryo.

The present video demonstrates a method which takes advantage of the combination of electroporation and confocal microscopy to perform live imaging on individual neural progenitor cells in the developing zebrafish forebrain. In vivo analysis of the development of forebrain neural progenitor cells at a clonal level can be achieved in this way.

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 ...

Investigating Social Cognition in Infants and Adults Using Dense Array Electroencephalography (dEEG)

Dense array electroencephalography (dEEG), which provides a non-invasive window for measuring brain activity and a temporal resolution unsurpassed by any other current brain imaging technology1,2, is being used increasingly in the study of social cognitive functioning in infants and adults. While dEEG is enabling researchers to examine brain activity patterns with unprecedented levels of sensitivity, conventional EEG recording systems continue to face certain limitations, including 1) poor spatial resolution and source localization3,4,2) the physical discomfort for test subjects of enduring the individual application of numerous electrodes to the surface of the scalp, and 3) the complexity for researchers of learning to use multiple software packages to collect and process data. Here we present an overview of an established methodology that represents a significant improvement on conventional methodologies for studying EEG in infants and adults. Although several analytical software techniques can be used to establish indirect indices of source localization to improve the spatial resolution of dEEG, the HydroCel Geodesic Sensor Net (HCGSN) by Electrical Geodesics, Inc. (EGI), a dense sensory array that maintains equal distances among adjacent recording electrodes on all surfaces of the scalp, further enhances spatial resolution4,5,6 compared to standard dEEG systems. The sponge-based HCGSN can be applied rapidly and without scalp abrasion, making it ideal for use with adults7,8, children9,10,11, and infants12, in both research and clinical4,5,6,13,14,15 settings. This feature allows for considerable cost and time savings by decreasing the average net application time compared to other dEEG systems. Moreover, the HCGSN includes unified, seamless software applications for all phases of data, greatly simplifying the collection, processing, and analysis of dEEG data.
The HCGSN features a low-profile electrode pedestal, which, when filled with electrolyte solution, creates a sealed microenvironment and an electrode-scalp interface. In all Geodesic dEEG systems, EEG sensors detect changes in voltage originating from the participant's scalp, along with a small amount of electrical noise originating from the room environment. Electrical signals from all sensors of the Geodesic sensor net are received simultaneously by the amplifier, where they are automatically processed, packaged, and sent to the data-acquisition computer (DAC). Once received by the DAC, scalp electrical activity can be isolated from artifacts for analysis using the filtering and artifact detection tools included in the EGI software. Typically, the HCGSN can be used continuously for only up to two hours because the electrolyte solution dries out over time, gradually decreasing the quality of the scalp-electrode interface.
In the Parent-Infant Research Lab at the University of Toronto, we are using dEEG to study social cognitive processes including memory, emotion, goals, intentionality, anticipation, and executive functioning in both adult and infant participants.

Investigating Social Cognition in Infants and Adults Using Dense Array Electroencephalography dEEG

Dense array electroencephalography is being used increasingly to study social cognitive functions in infants and adults. Here we present an established methodology that represents a significant improvement on conventional methodologies for studying EEG in infants and adults.

调查婴儿和成人使用密集阵脑电图的社会认知（ D脑电图）

Dense array electroencephalography (dEEG), which provides a non-invasive window for measuring brain activity and a temporal resolution unsurpassed by any ...

Local and Global Methods of Assessing Thermal Nociception in Drosophila Larvae

In this article, we demonstrate assays to study thermal nociception in Drosophila larvae. One assay involves spatially-restricted (local) stimulation of thermal nociceptors1,2 while the second involves a wholesale (global) activation of most or all such neurons3. Together, these techniques allow visualization and quantification of the behavioral functions of Drosophila nociceptive sensory neurons.
The Drosophila larva is an established model system to study thermal nociception, a sensory response to potentially harmful temperatures that is evolutionarily conserved across species1,2. The advantages of Drosophila for such studies are the relative simplicity of its nervous system and the sophistication of the genetic techniques that can be used to dissect the molecular basis of the underlying biology4-6 In Drosophila, as in all metazoans, the response to noxious thermal stimuli generally involves a "nocifensive" aversive withdrawal to the presented stimulus7. Such stimuli are detected through free nerve endings or nociceptors and the amplitude of the organismal response depends on the number of nociceptors receiving the noxious stimulus8. In Drosophila, it is the class IV dendritic arborization sensory neurons that detect noxious thermal and mechanical stimuli9 in addition to their recently discovered role as photoreceptors10. These neurons, which have been very well studied at the developmental level, arborize over the barrier epidermal sheet and make contacts with nearly all epidermal cells11,12. The single axon of each class IV neuron projects into the ventral nerve cord of the central nervous system11 where they may connect to second-order neurons that project to the brain.
Under baseline conditions, nociceptive sensory neurons will not fire until a relatively high threshold is reached. The assays described here allow the investigator to quantify baseline behavioral responses or, presumably, the sensitization that ensues following tissue damage. Each assay provokes distinct but related locomotory behavioral responses to noxious thermal stimuli and permits the researcher to visualize and quantify various aspects of thermal nociception in Drosophila larvae. The assays can be applied to larvae of desired genotypes or to larvae raised under different environmental conditions that might impact nociception. Since thermal nociception is conserved across species, the findings gleaned from genetic dissection in Drosophila will likely inform our understanding of thermal nociception in other species, including vertebrates.

Local and Global Methods of Assessing Thermal Nociception in Drosophila Larvae

In this article, we demonstrate assays to study thermal nociception in Drosophila larvae. One assay involves spatially-restricted (local) stimulation of thermal nociceptors1,2 while the second involves a wholesale (global) activation of most or all such neurons3. Together, these techniques allow visualization and quantification of the behavioral functions of Drosophila nociceptive sensory neurons.

局部和全局的方法评估热痛觉<em&gt;果蝇</em&gt;幼虫

In this article, we demonstrate assays to study thermal nociception in Drosophila larvae. One assay involves spatially-restricted (local) stimulation of ...

Dual Electrophysiological Recordings of Synaptically-evoked Astroglial and Neuronal Responses in Acute Hippocampal Slices

Astrocytes form together with neurons tripartite synapses, where they integrate and modulate neuronal activity. Indeed, astrocytes sense neuronal inputs through activation of their ion channels and neurotransmitter receptors, and process information in part through activity-dependent release of gliotransmitters. Furthermore, astrocytes constitute the main uptake system for glutamate, contribute to potassium spatial buffering, as well as to GABA clearance. These cells therefore constantly monitor synaptic activity, and are thereby sensitive indicators for alterations in synaptically-released glutamate, GABA and extracellular potassium levels. Additionally, alterations in astroglial uptake activity or buffering capacity can have severe effects on neuronal functions, and might be overlooked when characterizing physiopathological situations or knockout mice. Dual recording of neuronal and astroglial activities is therefore an important method to study alterations in synaptic strength associated to concomitant changes in astroglial uptake and buffering capacities. Here we describe how to prepare hippocampal slices, how to identify stratum radiatum astrocytes, and how to record simultaneously neuronal and astroglial electrophysiological responses. Furthermore, we describe how to isolate pharmacologically the synaptically-evoked astroglial currents.

The preparation of acute brain slices from isolated hippocampi, as well as the simultaneous electrophysiological recordings of astrocytes and neurons in stratum radiatum during stimulation of schaffer collaterals is described. The pharmacological isolation of astroglial potassium and glutamate transporter currents is demonstrated.

双突触诱发的星形胶质细胞和神经元的反应，在急性海马脑片电生理记录

Astrocytes form together with neurons tripartite synapses, where they integrate and modulate neuronal activity. Indeed, astrocytes sense neuronal inputs ...

Stereotaxic Injection of a Viral Vector for Conditional Gene Manipulation in the Mouse Spinal Cord

Intraparenchymal injection of a viral vector enables conditional gene manipulation in distinct populations of neurons or particular regions of the central nervous system. We demonstrate a stereotaxic injection technique that allows targeted gene expression or silencing in the dorsal horn of the mouse spinal cord. The surgical procedure is brief. It requires laminectomy of a single vertebra, providing for quick recovery of the animal and unimpaired motility of the spine. Controlled injection of a small vector suspension volume at low speed and use of a microsyringe with beveled glass cannula minimize the tissue lesion. The local immune response to the vector depends on the intrinsic properties of the virus employed; in our experience, it is minor and short-lived when a recombinant adeno-associated virus is used. A reporter gene such as enhanced green fluorescent protein facilitates monitoring spatial distribution of the vector, and the efficacy and cellular specificity of the transfection.

Viral vectors allow for targeted gene manipulation. We demonstrate a method for conditional gene expression or ablation in the mouse spinal cord, using stereotaxic injection of a viral vector into the dorsal horn, a prominent site of synaptic contact between primary somatosensory afferents and neurons of the central nervous system.

立体定向注射病毒载体的基因操作在小鼠脊髓条件

Intraparenchymal injection of a viral vector enables conditional gene manipulation in distinct populations of neurons or particular regions of the central ...

In Vivo Electrophysiological Measurements on Mouse Sciatic Nerves

Electrophysiological studies allow a rational classification of various neuromuscular diseases and are of help, together with neuropathological techniques, in the understanding of the underlying pathophysiology1. Here we describe a method to perform electrophysiological studies on mouse sciatic nerves in vivo.
The animals are anesthetized with isoflurane in order to ensure analgesia for the tested mice and undisturbed working environment during the measurements that take about 30 min/animal. A constant body temperature of 37 &#176;C is maintained by a heating plate and continuously measured by a rectal thermo probe2. Additionally, an electrocardiogram (ECG) is routinely recorded during the measurements in order to continuously monitor the physiological state of the investigated animals.
Electrophysiological recordings are performed on the sciatic nerve, the largest nerve of the peripheral nervous system (PNS), supplying the mouse hind limb with both motoric and sensory fiber tracts. In our protocol, sciatic nerves remain in situ and therefore do not have to be extracted or exposed, allowing measurements without any adverse nerve irritations along with actual recordings. Using appropriate needle electrodes3 we perform both proximal and distal nerve stimulations, registering the transmitted potentials with sensing electrodes at gastrocnemius muscles. After data processing, reliable and highly consistent values for the nerve conduction velocity (NCV) and the compound motor action potential (CMAP), the key parameters for quantification of gross peripheral nerve functioning, can be achieved.

In Vivo Electrophysiological Measurements on Mouse Sciatic Nerves

Measurements of nerve conduction properties in vivo exemplify a powerful tool to characterize various animal models of neuromuscular diseases. Here, we present an easy and reliable protocol by which electrophysiological analysis on sciatic nerves of anesthetized mice can be performed.

在对小鼠坐骨神经体内的电测量

Electrophysiological studies allow a rational classification of various neuromuscular diseases and are of help, together with neuropathological ...

Novel Assay for Cold Nociception in Drosophila Larvae

How organisms sense and respond to noxious temperatures is still poorly understood. Further, the mechanisms underlying sensitization of the sensory machinery, such as in patients experiencing peripheral neuropathy or injury-induced sensitization, are not well characterized. The genetically tractable Drosophila model has been used to study the cells and genes required for noxious heat detection, which has yielded multiple conserved genes of interest. Little is known however about the cells and receptors important for noxious cold sensing. Although, Drosophila does not survive prolonged exposure to cold temperatures (&#8804;10 &#186;C), and will avoid cool, preferring warmer temperatures in behavioral preference assays, how they sense and possibly avoid noxious cold stimuli has only recently been investigated.
Here we describe and characterize the first noxious cold (&#8804;10 &#186;C) behavioral assay in Drosophila. Using this tool and assay, we show an investigator how to qualitatively and quantitatively assess cold nociceptive behaviors. This can be done under normal/healthy culture conditions, or presumably in the context of disease, injury or sensitization. Further, this assay can be applied to larvae selected for desired genotypes, which might impact thermosensation, pain, or nociceptive sensitization. Given that pain is a highly conserved process, using this assay to further study&#160;thermal nociception will likely glean important understanding of pain processes in other species, including vertebrates.

Novel Assay for Cold Nociception in Drosophila Larvae

Here we demonstrate a novel assay to study cold nociception in Drosophila larvae. This assay utilizes a custom-built Peltier probe capable of applying a focal noxious cold stimulus and results in quantifiable cold-specific behaviors. This technique will allow further cellular and molecular dissection of cold nociception.

新的分析冷痛觉在<em&gt;果蝇</em&gt;幼虫

How organisms sense and respond to noxious temperatures is still poorly understood. Further, the mechanisms underlying sensitization of the sensory ...

3D Kinematic Analysis for the Functional Evaluation in the Rat Model of Sciatic Nerve Crush Injury

Compared to the Sciatic Functional Index (SFI), kinematic analysis is a more reliable and sensitive method for performing functional evaluations of sciatic nerve injury rodent models. In this protocol, we describe a novel kinematic analysis method that uses a three-dimensional (3D) motion capture apparatus for functional evaluations using a rat sciatic nerve crush injury model. First, the rat is familiarized with treadmill walking. Markers are then attached to the designated bone landmarks and the rat is made to walk on the treadmill at the desired speed. Meanwhile, the posterior limb movements of the rat are recorded using four cameras. Depending on the software used, marker tracings are created using both automatic and manual modes and the desired data are produced after subtle adjustments. This method of kinematic analysis, which uses a 3D motion capture apparatus, offers numerous advantages, including superior precision and accuracy. Many more parameters can be investigated during the comprehensive functional evaluations. This method has several shortcomings that require consideration: The system is expensive, can be complicated to operate, and may produce data deviations due to skin shifting. Nevertheless, kinematic analysis using a 3D motion capture apparatus is useful for performing functional anterior and posterior limb evaluations. In the future, this method may become increasingly useful for generating accurate assessments of various traumas and diseases.

We introduce a kinematic analysis method that uses a three-dimensional motion capture apparatus containing four cameras and data processing software for performing functional evaluations during fundamental research involving rodent models.

坐骨神经粉碎损伤大鼠模型中功能评估的3D运动学分析

Compared to the Sciatic Functional Index (SFI), kinematic analysis is a more reliable and sensitive method for performing functional evaluations of ...

Preparation of Acute Human Hippocampal Slices for Electrophysiological Recordings

Epilepsy affects about 1% of the world population and leads to a severe decrease in quality of life due to ongoing seizures as well as high risk for sudden death. Despite an abundance of available treatment options, about 30% of patients are drug-resistant. Several novel therapeutics have been developed using animal models, though the rate of drug-resistant patients remains unaltered. One of probable reasons is the lack of translation between rodent models and humans, such as a weak representation of human pharmacoresistance in animal models. Resected human brain tissue as a preclinical evaluation tool has the advantage to bridge this translational gap. Described here is a method for high quality preparation of human hippocampal brain slices and subsequent stable induction of epileptiform activity. The protocol describes the induction of burst activity during application of 8 mM KCl and 4-aminopyridin. This activity is sensitive to established AED lacosamide or novel antiepileptic candidates, such as dimethylethanolamine (DMEA). In addition, the method describes induction of seizure-like events in CA1 of human hippocampal brain slices by reduction of extracellular Mg2+ and application of bicuculline, a GABAA receptor blocker. The experimental set-up can be used to screen potential antiepileptic substances for their effects on epileptiform activity. Furthermore, mechanisms of action postulated for specific compounds can be validated using this approach in human tissue (e.g., using patch-clamp recordings). To conclude, investigation of vital human brain tissue ex vivo (here, resected hippocampus from patients suffering from temporal lobe epilepsy) will improve the current knowledge of physiological and pathological mechanisms in the human brain.

The presented protocol describes the transport and preparation of resected human hippocampal tissue with the ultimate goal to use vital brain slices as a preclinical evaluation tool for potential antiepileptic substances.

为电子生理记录准备急性人类河马切片

Epilepsy affects about 1% of the world population and leads to a severe decrease in quality of life due to ongoing seizures as well as high risk for ...