我们要感谢她的支持莉娅McAleer在本次实验和她的帮助下进行的海报介绍​​，在2009年神经科学会议和本视频制作协会，以​​及与穆罕默德阿卜杜拉和Allison阿伦斯。我们也感谢其在数据收集和分析本实验的援助罗茜马多克斯，Chavana雷切尔和琳达鞠。这个项目是由NIH / NIDA的格兰特3R01DA014640 - 05S1（CLD）的瓦戈纳中心，对酒精和成瘾问题研究琼斯院士和美国国立卫生研究院/ NIAA培训资助AA07471（AAF）的支持。可卡因盐酸慷慨提供由NIDA的药品库存和供应及控制程序。

*=最重要的步骤<ol><li>精心处理的雄性Sprague - Dawley大鼠进行培训，以杠杆压与食物奖励（45毫克蔗糖颗粒;生物服务，弗伦奇敦，新泽西州）。 </li><li> *静脉导管的构造，不锈钢套管（塑料之一，弗吉尼亚州）和硅橡胶管。大鼠接受一个颈静脉导管手术过程，让药物的自我管理。麻醉组成的氧气的混合物（0.8升/分钟;公司Airgas西南，科珀斯克里斯蒂，TX）和异氟醚（2.5-4％; AErrane，百特医疗用品，迪尔菲尔德，IL）通过气体输送系统（VetEquip公司，普莱森的交付，CA）。手术后，导管通畅，维持每天0.1毫升0.9％生理盐水，肝素和Timentin（抗生素）解决方案的冲洗。庆大霉素，外用抗生素，适用于头盖骨和胸部切口，以防止感染。所有的手术过程中遵循无菌动物手术的指导方针。蒸压外科用品和仪器在化学溶液和热珠灭菌消毒。植入导管冲洗，在制备过程中的甲醇，水和空气，然后沐浴在手术当天的70％的乙醇溶液。大鼠手术准备剃的头，胸部，其次应用了优碘溶液。手术后的大鼠进行了监测几个小时才开始返回到动物的殖民地。 </li><li>可卡因（NIDA的药品库存，供应和控制程序; RTI国际研究三角园，数控），在这个实验是在适当的剂量浓度溶解在生理盐水溶液（0.9％），根据动物重量​​。可卡因的解决方案，然后通过0.2微米的无菌注射器过滤器每天使用前过滤。 </li><li> *该仪器是构造出两个单杆操作性配备了房子和刺激的灯光和photobeams 3集（MED Associates公司，圣奥尔本斯，VT）的钱伯斯（28 × 22 × 21厘米）。钱伯斯联手构造的有机玻璃巷（21 × 25 × 25厘米）。胡同由两个黑色与白色条纹的墙壁和两个金属墙壁，一个明确的有机玻璃顶部，和白色的树脂玻璃地板。两个操作性商会钢格杆（4.8毫米）地板安装可拆卸的金属笼底。感官线索都被放置在操作性商会，创造出独特的，区分环境。视觉线索由白色或黑色的感觉材料的正面和背面的墙壁，并在笼顶。嗅觉线索石油为基础的气味（玫瑰或肉桂）饱和棒室地板下的棉花球。 </li><li>实验过程中发生多天，与基线偏好测试（2天），其次是药物的自我管理会议（8或16天），开始和结束一个条件性位置偏爱（CPP）的测试（1天）。 </li><li> *在基线偏好和CPP测试程序，动物也有获得两个操作性商会。以上设备安装一台摄像机记录每个分庭老鼠的出入口。商会内的Photobeams检测自发活动指数的光束，破损。在药物自我管理会话，动物仅限于一个单一的操作性室，而杠杆的反应和自发活动（例如，photobeam破损）数据与一个医学奔腾100兆赫配备MED - PC软件（MED Associates公司的电脑记录，圣奥尔本斯，佛蒙特州）。 </li><li>连续两天收集基线偏好测量。首先，老鼠是摆在中心巷，访问了20分钟的时间内既操作性商会。后立即放置在会议厅内，摄像机，MED - PC的软件程序和计时器同时启动。基线偏好数据评估过程的录像收看。 </li><li> *药物的自我管理的会议后展开的最后一个基准测试的日子。感官线索（例如，白色或黑色的墙面和玫瑰或肉桂香味）和面板封锁访问有机玻璃中心巷放入前动物室。要启动会话，动物置于内室，而从装药注射器管连接到动物的导管入口。操作性室门关闭后，MED - PC程序和定时器的启动和可卡因的访问是为1小时。注入0.1毫升（0.75毫克/公斤/损伤）的可卡因或生理盐水溶液中的每一杆的响应结果。在每个自我管理会议结束时，老鼠是从药管断开，从室中删除，并放置在家里笼。 </li><li>在过去的自我管理会议后的一天，动物接受一个条件性位置偏爱（CPP）的测试。为基线测量（见7）的过程是相同的。 </li><li> *数据评估：盲组分配实验者确定的金额通过观看录像带，以评估车厢之间的入口和出口，15分钟内（开始后5分钟内设备的位置），当时的动物花费在每个车厢。同样的程序是用来确定基准的偏好和CPP测试成绩。 </li><li> *数据分析：基线测量和调节后CPP分数之间的差异，以确定药物调节作用。计算差异分数减去在生理盐水配对车厢花费的时间量从量的时间花费在药物配对舱在CPP测试药物配对车厢（秒数） - （生理盐水中的秒数配对室）。可以执行差异得分的变化，从基线到测试阶段的统计分析，以确定药物空调的效果。 </li></ol> CPP的分数也可以表示为CPP的差值的百分比（％）。在本研究中，首先计算CPP差异分数％％的时间花费在每个操作性室使用公式确定：（在药物的时间（或生理盐水）在每个分庭室/总时间）× 100 =％条件性位置偏爱（CPP）。不同的得分百分比由公式：（药物配对％） - （生理盐水配对％）= CPP的差异分数％。 <img src="/files/ftp_upload/1998/1998fig1.jpg" alt="图1" /> 图1。关系可卡因摄入量和条件性位置偏爱之间的皮尔逊相关性分析，确定一个显着的正面相关性的动物共有可卡因的钢筋杆的响应和CPP的差异分数％之间的八个会话（例如，长期组的自我管理可卡因 的 ;。p &lt;0.05），但不适合在短期组（例如，4可卡因会）的动物。开放的钻石代表可卡因长期（N = 9）和实心圆代表可卡因短期（N = 8）组单独的数据点。最佳拟合线是虚线为长期和短期数据点的固体。

<ol>
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操作性静脉注射毒品的自我管理和地方调节程序是可靠和有效的模型，为研究药物依赖和成瘾1 2 3的神经生物学基础。这两种方法被广泛应用于临床前药物滥用研究，能够测量4个滥用药物的补强性能。然而，这两种方法有缺点，这里介绍的新仪器和方法改进后，5。 CPP的传统程序的一个大缺点是药品监督管理局非队伍的模式。这种药物摄入模式比自我管理6 7 8 9 10 11不同的行为和神经化学的成果，是不符合人类吸毒的经验。此外，不像药物自我管理程序，CPP的范例是无法衡量逐步改变药物摄入量增加反映在药物的动机，一个公认的转折点，在娱乐性药物的使用不可控制的吸毒开关的的。然而，药物自我管理行为，以及解释性的限制。例如，回应率是经常被用来推断奖励价值，但可以直接影响自我管理的药物，药物的激励效果无关的肌肉运动的影响。此外，增强药物的反应也可以影响的存在或缺乏药物输液相关的刺激 12 。 可卡因是已知的老鼠容易自我管理，并产生各种不同的剂量和管理2 13月14日航线强大的条件性位置偏好。从目前的研究结果支持了以前的报告药物加固模型。此外，可卡因增强杠杆反应和CPP得分之间显着正相关关系，确定在龙，而不是短期的可卡因集团。这些结果表明，地方调节和自我管理不一定是同构的奖励措施。举例来说，这是可以想象的短期暴露于可卡因后，CPP的反映急性加强与最初的娱乐性药物的使用通常看到的属性。另一方面，在更多的可卡因经验，可卡因空调加固水平的不断提高通信的摄入量升级大鼠可能表明药物敏感或有价值的药物作用，增强了某些人群的逐步变化。 此功能的方法，已超出评估的积极加强特效药物的实用工具。例如，厌恶药物的影响，不存在在最初使用药物可以延长药物暴露（如空调的地方厌恶，或CPA）出现，并会被检测出使用这种技术。其他用途包括为最敏感可卡因线索联想学习的亚群屏幕上的潜力，在可卡因的动机的行为，以评估经验介导的变化，并检测空调持久的药物戒毒和/或线索复职期间的影响。总之，本仪器和方法是时间和成本效益，可用于研究一个主题，有关滥用药物的广泛，并提供比cpp或自我管理的方法单独的实验设计更具灵活性。

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<td>Operant Conditioning Behavior (Drug Self-administration) Test Package for Rat</td>
<td>&nbsp;</td>
<td><a href="http://www.med-associates.com/">Med Associates, Inc</a></td>
<td>MED-008-CT-B1</td>
<td>&nbsp;</td>
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<td>Infrared Source and Detector (Photobeams)</td>
<td>&nbsp;</td>
<td><a href="http://www.med-associates.com/">Med Associates, Inc</a></td>
<td>ENV-253SD ENV-253</td>
<td>&nbsp;</td>
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<td>Med PC Software</td>
<td>&nbsp;</td>
<td><a href="http://www.med-associates.com/">Med Associates, Inc</a></td>
<td>SOF-735</td>
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<td>Single speed syringe pump</td>
<td>&nbsp;</td>
<td>Razel Scientific Instruments</td>
<td>Model R-E</td>
<td>&nbsp;</td>
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<td>45 mg sucrose pellets</td>
<td>&nbsp;</td>
<td>Bio-Serv</td>
<td>F0042</td>
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<td>Plastics One</td>
<td>C313G-5UP</td>
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<td>Cocaine</td>
<td>&nbsp;</td>
<td>RTI International</td>
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novel apparatus and method for drug reinforcement

动物模型的钢筋已被证明是用于了解基本药物成瘾的神经生物学机制。操作性药物自我管理和条件性位置偏爱（CPP）的程序是豪气用在动物研究模型药物加固，消费，并在人类成瘾的各个组成部分。在这项研究中，我们使用了新的方法来研究大鼠药物加固相结合，传统的CPP和自我管理方法。我们组装的仪器使用两个MED副操作性商会，感官刺激，和一个有机玻璃构造的中立区。这些修改使我们的实验，包括药物摄入的动机方面，通过自我管理和无毒的药物/ CUE空调强度与CPP测试评估。在我们的实验中，老鼠的自我管理可卡因在四个（例如，“短期”）或8个（0.75毫克/公斤/损伤，IV）（例如，“长期”）交替天，在会议操作性的环境中含有独特的感官线索（例如，嗅觉和视觉）。在隔天，在其他的（不同的线索）操作性的环境，生理盐水可用于自我注射（0.1毫升，IV）。二十四小时后，最后self-administration/cue-pairing会议，进行了一个CPP测试。与典型的CPP结果相一致，有一个显着的偏好与可卡因的自我管理的室。此外，发生在动物的长期实验，CPP的幅度和可卡因钢筋杆反应之间的一个显着的正相关关系。总之，本仪器和方法是时间和成本效益，可用于研究一个主题，有关滥用药物的广泛，并提供比cpp或自我管理的方法单独的实验设计更具灵活性。

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Novel Apparatus and Method for Drug Reinforcement

操作性药物自我管理和条件性位置偏爱（CPP）的程序是豪气用于研究模型药物加固，消费，并在人类成瘾的各个组成部分。在这份报告中，我们结合传统作为一种新的方法来研究药物加固和成瘾大鼠的CPP和自我管理的方法。

新型仪器和药物加固方法

Animal models of reinforcement have proven to be useful for understanding the neurobiological mechanisms underlying drug addiction.  Operant drug ...

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19.0K 次观看.  University of Texas at Austin. 操作性药物自我管理和条件性位置偏爱（CPP）的程序是豪气用于研究模型药物加固，消费，并在人类成瘾的各个组成部分。在这份报告中，我们结合传统作为一种新的方法来研究药物加固和成瘾大鼠的CPP和自我管理的方法。

视频: Novel Apparatus and Method for Drug Reinforcement

A Novel Method for Assessing Proximal and Distal Forelimb Function in the Rat: the Irvine, Beatties and Bresnahan (IBB) Forelimb Scale

Several experimental models of cervical spinal cord injury (SCI) have been developed recently to assess the consequences of damage to this level of the spinal cord (Pearse et al., 2005, Gensel et al., 2006, Anderson et al., 2009), as the majority of human SCI occur here (Young, 2010; www.sci-info-pages.com). Behavioral deficits include loss of forelimb function due to damage to the white matter affecting both descending motor and ascending sensory systems, and to the gray matter containing the segmental circuitry for processing sensory input and motor output for the forelimb. Additionally, a key priority for human patients with cervical SCI is restoration of hand/arm function (Anderson, 2004). Thus, outcome measures that assess both proximal and distal forelimb function are needed. Although there are several behavioral assays that are sensitive to different aspects of forelimb recovery in experimental models of cervical SCI (Girgis et al., 2007, Gensel et al., 2006, Ballerman et al., 2001, Metz and Whishaw, 2000, Bertelli and Mira, 1993, Montoya et al., 1991, Whishaw and Pellis, 1990), few techniques provide detailed information on the recovery of fine motor control and digit movement.
The current measurement technique, the Irvine, Beatties and Bresnahan forelimb scale (IBB), can detect recovery of both proximal and distal forelimb function including digit movements during a naturally occurring behavior that does not require extensive training or deprivation to enhance motivation. The IBB was generated by observing recovery after a unilateral C6 SCI, and involves video recording of animals eating two differently shaped cereals (spherical and doughnut) of a consistent size. These videos were then used to assess features of forelimb use, such as joint position, object support, digit movement and grasping technique.
The IBB, like other forelimb behavioral tasks, shows a consistent pattern of recovery that is sensitive to injury severity. Furthermore, the IBB scale could be used to assess recovery following other types of injury that impact normal forelimb function.

A Novel Method for Assessing Proximal and Distal Forelimb Function in the Rat: the Irvine, Beatties and Bresnahan IBB Forelimb Scale

Here we will describe a rodent behavioral assay that can detect recovery of both proximal and distal forelimb function including digit movements during a naturally occurring behavior that does not require extensive training or deprivation to enhance motivation.

一个评估在大鼠近端和远端的前肢功能的新方法：欧文，Beatties和布雷斯纳汉（IBB）前肢规模

Several experimental models of cervical spinal cord injury (SCI) have been developed recently to assess the consequences of damage to this level of the ...

科研

神经科学

A Novel Method of Drug Administration to Multiple Zebrafish (Danio rerio) and the Quantification of Withdrawal

Anxiety testing in zebrafish is often studied in combination with the application of pharmacological substances. In these studies, fish are routinely netted and transported between home aquaria and dosing tanks. In order to enhance the ease of compound administration, a novel method for transferring fish between tanks for drug administration was developed. Inserts that are designed for spawning were used to transfer groups of fish into the drug solution, allowing accurate dosing of all fish in the group. This increases the precision and efficiency of dosing, which becomes very important in long schedules of repeated drug administration. We implemented this procedure for use in a study examining the behavior of zebrafish in the light/dark test after administering ethanol with differing 21 day schedules. In fish exposed to daily-moderate amounts of alcohol there was a significant difference in location preference after 2 days of withdrawal when compared to the control group. However, a significant difference in location preference in a group exposed to weekly-binge administration was not observed. 
This protocol can be generalized for use with all types of compounds that are water-soluble and may be used in any situation when the behavior of fish during or after long schedules of drug administration is being examined. The light/dark test is also a valuable method of assessing withdrawal-induced changes in anxiety.

A Novel Method of Drug Administration to Multiple Zebrafish Danio rerio and the Quantification of Withdrawal

A novel method for reducing variability when exposing fish to drugs is explained. Fish exposed to various patterns of ethanol exposure were found to have altered anxiety levels during withdrawal in a light/dark scototaxic assay.

药品监督管理局多个斑马鱼的新方法（<em&gt;斑马鱼</em&gt;）和退出的量化

Anxiety testing in zebrafish is often studied in combination with the application of pharmacological substances. In these studies, fish are routinely ...

Assessment of Neuronal Viability Using Fluorescein Diacetate-Propidium Iodide Double Staining in Cerebellar Granule Neuron Culture

Primary cultured Cerebellar Granule Neurons (CGNs) have been widely used as an in vitro model in neuroscience and neuropharmacology research. However, the co-existence of glial cells and neurons in CGN culture might lead to biases in the accurate assessment of neuronal viability. Fluorescein diacetate (FDA) and Propidium Iodide (PI) double staining has been used to measure cell viability by simultaneously evaluating the viable and dead cells. We used FDA-PI double staining to improve the sensitivities of the colorimetric assays and to evaluate neuronal viability in CGNs. Furthermore, we added blue fluorescent DNA stains (e.g., Hoechst) to improve the accuracy. This protocol describes how to improve the accuracy of assessment of neuronal viability by using these methods in CGN culture. Using this protocol, the number of glial cells can be excluded by using fluorescence microscopy. A similar strategy can be applied to distinguish the unwanted glial cells from neurons in various mixed cell cultures, such as primary cortical culture and hippocampal culture.

This protocol describes how to accurately measure neuronal viability using Fluorescein diacetate (FDA) and Propidium Iodide (PI) double staining in cultured cerebellar granule neurons, a primary neuronal culture used as an in vitro model in neuroscience and neuropharmacology research.

使用荧光素二乙酸酯 - 碘化丙啶双重染色在小脑颗粒神经元培养中评估神经元活力

Primary cultured Cerebellar Granule Neurons (CGNs) have been widely used as an in vitro model in neuroscience and neuropharmacology research. However, the ...

Recording EEG in Freely Moving Neonatal Rats Using a Novel Method

EEG is a useful method to detect electrical activity in the brain. Moreover, it is a widely used diagnostic tool for various neurological conditions, such as epilepsy and neurodegenerative disorders. However, it is technically difficult to obtain EEG recordings in neonates as it requires specialized handling and great care. Here, we present a novel method to record EEG in neonatal rat pups (P8-P15). We designed a simple and reliable electrode using computer pin loci; it can be easily implanted into the skull of a rat pup to record high-quality EEG signals in the normal and epileptic brain. Pups were given an intraperitoneal (i.p.) injection of the neurotoxin kainic acid (KA) to induce epileptic seizures. The surgical implantation performed in this procedure is less expensive than other EEG procedures for neonates. This method allows one to record high-quality and stable EEG signals for more than 1 week. Furthermore, this procedure can also be applied to adult rats and mice to study epilepsy or other neurological disorders.

Here, we introduce a novel technique designed to record electroencephalography (EEG) in freely moving neonatal epileptic pups and describe its procedures, features, and applications. This method allows one to record EEG for more than 1 week.

使用新颖的方法在自由移动的新生大鼠中记录脑电图

EEG is a useful method to detect electrical activity in the brain. Moreover, it is a widely used diagnostic tool for various neurological conditions, such ...

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 ...

A Novel Method to Model Chronic Traumatic Encephalopathy in Drosophila

Chronic Traumatic Encephalopathy (CTE) is an established neurodegenerative disease that is closely associated with exposure to repetitive mild Traumatic Brain Injury (mTBI). The mechanisms responsible for its complex pathological changes remain largely elusive, despite a recent consensus to define the neuropathological criteria. Here, we describe a novel method to develop a model of CTE in Drosophila melanogaster&#160;(Drosophila&#160;)&#160;in an attempt to identify the key genes and pathways that lead to the characteristic hyperphosphorylated tau accumulation and neuronal death in the brain. Adjustable-strength impacts to inflict mild closed injury are delivered directly to the fly head, subjecting the head to rapid acceleration and deceleration. Our method eliminates the potential problems inherent with other Drosophila mTBI models (e.g.,animal death might be induced by damage to other parts of the body or to internal organs). The less labor- and cost-intensive animal care, short life span, and extensive genetic tools make the fruit fly ideal to study CTE pathogenesis and make it possible to perform large-scale, genome-wide forward genetic and pharmacological screens. We anticipate that the ongoing characterization of the model will generate important mechanistic insights on disease prevention and therapeutic approaches.

A Novel Method to Model Chronic Traumatic Encephalopathy in Drosophila

Here, we describe a new approach to inflict closed-head traumatic brain injury in Drosophila melanogaster. Our method has the advantage of directly delivering repetitive impacts with adjustable strength to the head alone. Further exploration of the invertebrate system will help to illuminate the pathogenesis of chronic traumatic encephalopathy.

慢性创伤性脑病模型的一种新方法<em&gt;果蝇</em

Chronic Traumatic Encephalopathy (CTE) is an established neurodegenerative disease that is closely associated with exposure to repetitive mild Traumatic ...

Assay Development for High Content Quantification of Sod1 Mutant Protein Aggregate Formation in Living Cells

Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disease that can be caused by inherited mutations in the gene encoding copper-zinc superoxide dismutase 1 (SOD1). The structural instability of SOD1 and the detection of SOD1-positive inclusions in familial-ALS patients supports a potential causal role for misfolded and/or aggregated SOD1 in ALS pathology. In this study, we describe the development of a cell-based assay designed to quantify the dynamics of SOD1 aggregation in living cells by high content screening approaches. Using lentiviral vectors, we generated stable cell lines expressing wild-type and mutant A4V SOD1 tagged with yellow fluorescent protein and found that both proteins were expressed in the cytosol without any sign of aggregation. Interestingly, only SOD1 A4V stably expressed in HEK-293, but not in U2OS or SH-SY5Y cell lines, formed aggregates upon proteasome inhibitor treatment. We show that it is possible to quantify aggregation based on dose-response analysis of various proteasome inhibitors, and to track aggregate-formation kinetics by time-lapse microscopy. Our approach introduces the possibility of quantifying the effect of ALS mutations on the role of SOD1 in aggregate formation as well as screening for small molecules that prevent SOD1 A4V aggregation.

We describe a method to quantify the aggregation of misfolded proteins. Our protocol details lentiviral induced stable cell line generation, automated confocal imaging, and image analysis of protein aggregates. As an illustrative application, we studied the effect of small molecules in promoting SOD1 aggregation in a time- and dose-dependent manner.

活细胞 Sod1 突变蛋白聚集物高含量定量测定方法的建立

Amyotrophic lateral sclerosis (ALS) is a fatal neurodegenerative disease that can be caused by inherited mutations in the gene encoding copper-zinc ...

Implantation of Chronic Silicon Probes and Recording of Hippocampal Place Cells in an Enriched Treadmill Apparatus

An important requisite for understanding brain function is the identification of behavior and cell activity correlates. Silicon probes are advanced electrodes for large-scale electrophysiological recording of neuronal activity, but the procedures for their chronic implantation are still underdeveloped. The activity of hippocampal place cells is known to correlate with an animal&#39;s position in the environment, but the underlying mechanisms are still unclear. To investigate place cells, here we describe a set of techniques which range from the fabrication of devices for chronic silicon probe implants to the monitoring of place field activity in a cue-enriched treadmill apparatus. A micro-drive and a hat are built by fitting and fastening together 3D-printed plastic parts. A silicon probe is mounted on the micro-drive, cleaned, and coated with dye. A first surgery is performed to fix the hat on the skull of a mouse. Small landmarks are fabricated and attached to the belt of a treadmill. The mouse is trained to run head-fixed on the treadmill. A second surgery is performed to implant the silicon probe in the hippocampus, following which broadband electrophysiological signals are recorded. Finally, the silicon probe is recovered and cleaned for reuse. The analysis of place cell activity in the treadmill reveals a diversity of place field mechanisms, outlining the benefit of the approach.

We describe the diverse steps to implant chronic silicon probes and to record place cells in mice that are running head-fixed on a cue-enriched treadmill apparatus.

富氧运动器械中慢性硅探针的植入及海马位细胞的记录

An important requisite for understanding brain function is the identification of behavior and cell activity correlates. Silicon probes are advanced ...

CMAP Scan MUNE (MScan) - A Novel Motor Unit Number Estimation (MUNE) Method

Like other methods for motor unit number estimation (MUNE), compound muscle action potential (CMAP) scan MUNE (MScan) is a non-invasive electrophysiologic method to estimate the number of functioning motor units in a muscle. MUNE is an important tool for the assessment of neuropathies and neuronopathies. Unlike most MUNE methods in use, MScan assesses all the motor units in a muscle, by fitting a model to a detailed stimulus-response curve, or CMAP scan. It thereby avoids the bias inherent in all MUNE methods based on extrapolating from a small sample of units. Like &#39;Bayesian MUNE,&#39;&#160;MScan analysis works by fitting a model, made up of motor units with different amplitudes, thresholds, and threshold variabilities, but the fitting method is quite different, and completed within five minutes, rather than several hours. The MScan off-line analysis works in two stages: first, a preliminary model is generated based on the slope and variance of the points in the scan, and second, this model is then refined by adjusting all the parameters to improve the fit between the original scan and scans generated by the model.
This new method has been tested for reproducibility and recording time on 22 amyotrophic lateral sclerosis (ALS) patients and 20 healthy controls, with each test repeated twice by two blinded physicians. MScan showed excellent intra- and inter-rater reproducibility with ICC values of &#62;0.98 and a coefficient of variation averaging 12.3 &#177; 1.6%. There was no difference in the intra-rater reproducibility between the two observers. Average recording time was 6.27 &#177; 0.27 min.
This protocol describes how to record a CMAP scan and how to use the MScan software to derive an estimate of the number and sizes of the functioning motor units. MScan is a fast, convenient, and reproducible method, which may be helpful in diagnoses and monitoring disease progression in neuromuscular disorders.

CMAP Scan MUNE MScan - A Novel Motor Unit Number Estimation MUNE Method

This protocol describes a new method to estimate the number of functioning motor units in a muscle, by fitting a model to a detailed stimulus-response curve of the compound muscle action potential. It is quick and easy to perform and analyze and has excellent reproducibility.

CMAP 扫描免疫性 (MScan)-一种新的电机单元数估计 (免疫性) 方法

Like other methods for motor unit number estimation (MUNE), compound muscle action potential (CMAP) scan MUNE (MScan) is a non-invasive electrophysiologic ...

Autoradiography as a Simple and Powerful Method for Visualization and Characterization of Pharmacological Targets

In vitro autoradiography aims to visualize the anatomical distribution of a protein of interest in tissue from experimental animals as well as humans. The method is based on the specific binding of a radioligand to its biological target. Therefore, frozen tissue sections are incubated with radioligand solution, and the binding to the target is subsequently localized by the detection of radioactive decay, for example, by using photosensitive film or phosphor imaging plates. Resulting digital autoradiograms display remarkable spatial resolution, which enables quantification and localization of radioligand binding in distinct anatomical structures. Moreover, quantification allows for the pharmacological characterization of ligand affinity by means of dissociation constants (Kd), inhibition constants (Ki) as well as the density of binding sites (Bmax) in selected tissues. Thus, the method provides information about both target localization and ligand selectivity. Here, the technique is exemplified with autoradiographic characterization of the high-affinity &#947;-hydroxybutyric acid (GHB) binding sites in mammalian brain tissue, with special emphasis on methodological considerations regarding the binding assay parameters, the choice of the radioligand and the detection method.

The method of autoradiography is routinely used to study binding of radioligands to tissue sections for determination of qualitative or quantitative pharmacology.

自体影像学作为一种简单而有力的药理靶标可视化和表征方法

In vitro autoradiography aims to visualize the anatomical distribution of a protein of interest in tissue from experimental animals as well as humans. The ...