这项工作是在大学医院的埃尔兰根 - 纽伦堡大学的跨学科研究中心临床研究（IZKF，项目E7）。

1。设置装配和软件编程<ol><li> D / A卡安装在个人计算机上（ 如 ：6229，美国国家仪器公司NI PCI），并把它连接到突破框（ 如 ：BNC-2110年，美国国家仪器公司），都应该至少支持一个输入和一个输出通道至少各44.1千赫的采样率。 </li><li>将突破盒的输出通过BNC电缆连接到声音放大器（ 如 ：AMP75宽带功率放大器，托马斯·沃尔夫）。 </li><li>安装一个红外摄像头（ 例如 ：大IP摄像机专业，GrandTec电子）在黑暗中的动物监测。 </li><li>安装一个集成开发环境（ 如 MATLAB）实施程序的流程图图1中给出的定义。负责从相应的作者可以得到免费的版本上运行的Matlab的2007年b。 </li><li>在一个隔音室安装一个扬声器上表。将扬声器TØ声音放大器。 </li><li>绝缘板的顶部安装一个压电传感器（ 如力传感器FSG15N1A，霍尼韦尔），支持有能力，有突破箱的输入通过BNC电缆连接到地面传感器信号。 </li><li>生成的测量腔的啮齿类动物的大小调整到由丙烯酸玻璃管来衡量（沙土鼠，例如 ：长度为15厘米，内径4.3厘米，外径4.8厘米）。修复篦带有网眼为0.5毫米的宽度，以前面的管和软垫的门的锁定机构（ 例如 ，钩）向后方。固定可以用热胶，门只的铰链和钩吊环被固定到所述管。 </li><li>将发泡塑料脚，适合测量管的尺寸，绝缘传感器板。呎应该支持下管的前端部和后端，并解除管传感器级别。确保压电传感器和测量室之间有轻微的接触。 </李&gt; <li>修正传感器板的测量室集中在扬声器前面和把麦克风声音控制（ 例如乙＆K型2669 /乙＆ķ类型4190连接到测量放大器乙＆ķ类型2610，所有：布乃尔和克亚尔）旁边的水平的动物的头部，以便它不会干扰与管。 </li><li>请注意，您的听力资料的质量将取决于您的音响系统的质量。在任何情况下，使用麦克风和测量放大器的频率传递函数确定你的系统之前，你的第一个实验，包括您的软件程序中纠正这个频率传递函数，使您的扬声器平坦的光谱输出。 </li><li>对准摄像头的设置，这样就可以监测动物的行为。 </li></ol> 2。行为听阈的测定（听力图） <ol><li>以从它的首页笼的动物，并把它头部先在管;失去了门。 </li><li>把管上的泡沫的脚和传感器。关闭任何光线和关闭的大门室。分庭本身是没有空调，但有温度，湿度和其他环境变量周围的实验室。由通风器的空气的变化是不可取的，由于感应噪声，但该室的体积是支持动物许多小时的氧。等待15分钟，让动物习惯的设置。的的驯化时间是有益的动物，因为它可以冷静下来按自己的步伐和习惯管。另一方面，没有看到任何的行为差异在驯化时间在几个会议，这表明不熟悉的动物的管和室，在实验前需要额外会议。 </li><li>启动程序和定义参数的刺激（亦参参考。5）：刺激由不同频率纯音quencies。必须提交的惊吓刺激的声压等级够高，重复性引起的惊吓反应。在我们的实验室中，我们使用强度105分贝SPL（刺激持续时间6毫秒，2毫秒的余弦平方的上升和下降斜）引起惊吓反应在长爪沙鼠。测试刺激前惊跳刺激的呈现进行测试的范围内，通常是从下面的听力阈值的水平远高于阈值和频率覆盖整个可听范围内的物种中，在不同的频率和强度。在每次试验测试的惊吓和刺激的频率和持续时间相匹配，惊吓和测试刺激刺激间的时间间隔设置为100毫秒。使用至少15重复与随机刺激间隔为10±2.5秒，为每个测试刺激的频率和强度的组合（参见图2A中 ，左）。可以呈现在随后的测试刺激随机或非然domized订单。如果您使用的是一个非随机的方法（ 例如 ，一个测试所有测试的频率固定的刺激程度）允许5分钟的恢复之间不同的刺激套。需要注意的是所确定的绝对阈值，将取决于随机的刺激，但可能相对于阈值的变化不会（ 如声损伤后，比照参考。5）。 </li><li>在分析数据之前，删除无效的试验数据集（ 例如 ，试验中的动物的惊吓刺激之前移动的比照图2B）。 </li><li>一个时间窗口内的第一个50毫秒的惊吓刺激后，计算响应的振幅（峰值到峰值的响应的第一最小值之间的第一最大）和响应延迟（时间从刺激开始响应发病的）每个单个试验。 </li><li>装一个功能完整的响应幅度设定一个频率的数据，对所有有效SINGL prestimulus强度排序为玻尔兹曼Ë试验。 50％的的玻尔兹曼功能7点，表明这种刺激频率的听力阈值。 </li></ol> 3。声损伤和听力障碍的量化<ol><li>准备氯胺酮 - 甲苯噻嗪麻醉，用盐酸氯胺酮的混合物：96毫克/公斤（氯胺酮Ratiopharm公司，Ratiopharm公司），甲苯噻嗪盐酸盐：4毫克/公斤（Rompun 2％，拜耳）硫酸阿托品：1毫克/千克（Atropinsulfat，贝朗Melsungen的AG）和生理的NaCl溶液（Berlin-Chemie公司的AG，柏林）的比例为9:1:2:8。 </li><li>用3毫升/公斤的麻醉皮下注入动物。等待，直到动物是深度麻醉（约5分钟，检查反射， 例如 ，使用踏板撤退反射）。在测量过程中，为了维持麻醉，不断注入麻醉溶液3毫升/千克/小时，使用注射泵的速度。控制适当的设备（ 如通过摄像机），呼吸的生命体征和保持动物的体温placi纳克在变暖垫。 </li><li>诱导的声创伤， 例如，使用一个响亮的115分贝SPL纯质的音调， 例如 ：2 kHz的75分钟。 </li><li>结束后的创伤，停止注射泵和在唤醒笼变暖的垫在一个安静的地方，让动物醒着。定期检查的觉醒阶段，如果生命体征稳定。将其饲养笼中的动物，只有当它是完全清醒的。让动物从麻醉（至少2天），其余在它的首页笼中恢复过来。 </li><li>执行2.1至2.6。比较之前和之后的声损伤的听力阈值，通过计算对每个频率的听力损失的百分比。实验结束后，所有无痛苦地安乐死的动物。 </li></ol> 4。试验声幻影感知（主观耳鸣） <ol><li>进行这些测量，之前和之后的声损伤。 </li><li>按照2.1到2.2，如果动物是不是已经在安装程序。 </li><li> A N红棕色的刺激范式可以用来测试啮齿动物的主观耳鸣。所有这些方法中的比率是一个沉默的间隙内的背景噪声的显着性检验。如果该间隙被认为由动物，它可以被用作测试刺激减少惊吓反应类似的步骤2中所述的。如果动物患有耳鸣（即是可能发展声创伤后的），这耳鸣将被感知沉默的间隙内，因此，使得该间隙那么突出。的惊吓反应效果上的差距，因此将减弱，耳鸣动物相比健康对照组（PPI降低，比照8）。这两个略有不同的协议知觉测试。 </li><li>在这里介绍的第一个主观性耳鸣范例（ 图2A，中心，比照9）使用以下参数为刺激：不惊人死不休的声音强度105分贝，频率从1到16 kHz，1个八度音阶，stimul我们长6毫秒，2毫秒余弦平方的上升和下降斜坡。出示一个白噪声背景50分贝SPL在实验过程中，带有或不带15毫秒间隙之前的惊吓刺激100毫秒;本的至少15个试验的每个频率和间隙条件。如果使用非随机的方式，允许5分钟的恢复之间的不同刺激套。使用不同的惊吓刺激频率将给出一个粗略的估计程度的耳鸣频率。 </li><li>主观耳鸣作为第二范式（ 图2A，右），你可以使用下面的惊吓刺激参数：声音强度105分贝SPL，双击刺激，每次点击的持续时间为0.1毫秒和0.1毫秒的点击次数之间具有倒方向，第二次点击相对于第一个。提出了一种带通滤波，噪音为50 dB SPL的背景高斯滤波器宽度0.5个八度，中心频率范围从1到16 kHz的八度音。目前这种噪声EI进一步带或不带一个15毫秒的间隙之前100毫秒的惊吓刺激;目前至少15个试验的每个频率和间隙条件。如果使用非随机的方式，允许5分钟的恢复之间的不同刺激套。使用不同的中心频率的带通的背景噪声将得到所感知的耳鸣频率的一个粗略的估计。 </li><li>按照2.4〜2.5;所有获取的数据与为每个测试的频率的基准的数据集和每个创伤条件， 即 ，之前和之后的声创伤正常化。此引用是纯音（参见2.3）没有任何prestimulus的惊吓刺激的反应幅度。的频率来确定可以由纯音或中心频率的带通滤波的噪声。计算的平均响应每个参考和规范每个计算出的响应的振幅将通过其参考。 </li><li>除以归一化响应的间隙条件[振幅计算PPIn通过每个测试频率的平均值的归一化无间隙状态。 </li><li>创伤后以百分比计算PPI的变化，每个测试频率。 </li></ol>动物的惊吓反应，很容易生成和分析。 图2B简要介绍了一个典型的结果的一种动物，与纯音105分贝SPL没有任何prestimulus 15次的刺激。大多数试验是有效的和无效的审判，很容易识别（试验标志着由红方）。响应的幅度和等待时间，只计算有效的试验。 图3A中给出一个典型的行为阈值改变。 2记载的方法，获得的听力图的一个示例性的动物之前给予（蓝色）和之后（红色）在2 kHz（黄色区域）的声创伤。特别是在2 kHz清晰的听力损失。可以是一种主观耳鸣知觉的反应可见在图3B中，示出与上述相同的动物的归一化的反应振幅示例性刺激一个八度的下方和上方的创伤，如4.5中所述。前对刺激的响应的比较的有和无间隙（蓝色）和创伤后（红色）允许一个可能的耳鸣知觉的解释。下面的创伤频率没有变化的响应模式可以发现的，而上面的创伤的影响的间隙消失后的创伤，表示在这个频率mispercept。 <img alt="图1" src="/files/ftp_upload/4433/4433fig1.jpg" /> 图1。计划用于收购行为的阈值和主观耳鸣数据流图 。请注意，这仅仅是一个简化的程序代码版本。缩写：GUI - 图形用户界面，ISI - 刺激间隔间。 图2。听觉惊跳反应（ASR）的刺激。一个计划使用三种不同的刺激方案。左图：前脉冲抑制（PPI），ASR没有任何纯音测试刺激前（绿色）不惊人死不休音（红色）测量的响应时间，以蓝色表示。中心板：间隙/纯音的惊吓刺激呈现的白噪声背景的不同频率的噪音范例。右侧面板间隙/噪音范式，点击惊吓刺激呈现带通滤波的中心频率不同的背景。B听觉惊跳的反应与阈值的范例没有任何prestimulus的刺激频率在1 kHz记录的15个试验示范。三次试验都算无效（红色方块）移动之前已经在动物的刺激发病。 &lt;IMG ALT =“图3”SRC =“/ files/ftp_upload/4433/4433fig3.jpg”/&gt; 图3。 ASR在一个动物的典型结果。行为阈值（红色）（蓝色）和的声损伤在2 kHz（黄色区域）。阈值的计算从对PPI调制ASR协议的Boltzmann函数作为阈值的转折点。听力损失在2千赫到66％以上，而较远的距离的创伤频率1，经常可以看到，即使提高听力阈值，B归一化响应幅值（空心圆：单独审判，实心圆手段，晶须需要注意的是：标准差）在间隙/噪音单击“ASR协议（4.5）1和4 kHz中心频率的刺激。响应排序条件进行试验，和不带间隙中的噪声之前和之后的创伤在2 kHz。只有在4千赫的间隙的效果消失的创伤后表示主观耳鸣知觉解决此频率。

<ol>
	<li>Brown, M., Irvine, D. R., Park, V. N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Perceptual+learning+on+an+auditory+frequency+discrimination+task+by+cats:+association+with+changes+in+primary+auditory+cortex.">Perceptual learning on an auditory frequency discrimination task by cats: association with changes in primary auditory cortex.</a> Cereb. Cortex. 14, 952-965 (2004).</li><li>Ohl, F. W., Scheich, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Learning-induced+plasticity+in+animal+and+human+auditory+cortex.">Learning-induced plasticity in animal and human auditory cortex.</a> Curr. Opin. Neurobiol. 15, 470-477 (2005).</li><li>Koch, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+neurobiology+of+startle.">The neurobiology of startle.</a> Prog. Neurobiol. 59, 107-128 (1999).</li><li>Larrauri, J., Schmajuk, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Prepulse+inhibition+mechanisms+and+cognitive+processes:+a+review+and+model.">Prepulse inhibition mechanisms and cognitive processes: a review and model.</a> EXS. 98, 245-278 (2006).</li><li>Walter, M., Tziridis, K., Ahlf, S., Schulze, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Context+dependent+auditory+thresholds+determined+by+brainstem+audiometry+and+prepulse+inhibition+in+Mongolian+gerbils.">Context dependent auditory thresholds determined by brainstem audiometry and prepulse inhibition in Mongolian gerbils.</a> Open Journal of Acoustics. 2, 34-49 (2012).</li><li>Gaese, B. H., Nowotny, M., Pilz, P. K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Acoustic+startle+and+prepulse+inhibition+in+the+Mongolian+gerbil.">Acoustic startle and prepulse inhibition in the Mongolian gerbil.</a> Physiol. Behav. 98, 460-466 (2009).</li><li>Fechter, L. D., Sheppard, L., Young, J. S., Zeger, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Sensory+threshold+estimation+from+a+continuously+graded+response+produced+by+reflex+modification+audiometry.">Sensory threshold estimation from a continuously graded response produced by reflex modification audiometry.</a> J. Acoust. Soc. Am. 84, 179-185 (1988).</li><li>Turner, J. G., Parrish, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Gap+detection+methods+for+assessing+salicylate-induced+tinnitus+and+hyperacusis+in+rats.">Gap detection methods for assessing salicylate-induced tinnitus and hyperacusis in rats.</a> Am. J. Audiol. 17, 185-192 (2008).</li><li>Campeau, S., Davis, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Fear+potentiation+of+the+acoustic+startle+reflex+using+noises+of+various+spectral+frequencies+as+conditioned+stimuli.">Fear potentiation of the acoustic startle reflex using noises of various spectral frequencies as conditioned stimuli.</a> Animal Learning &#38; Behavior. 20, 177-186 (1992).</li><li>Young, J. S., Fechter, L. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Reflex+inhibition+procedures+for+animal+audiometry:+a+technique+for+assessing+ototoxicity.">Reflex inhibition procedures for animal audiometry: a technique for assessing ototoxicity.</a> J. Acoust. Soc. Am. 73, 1686-1693 (1983).</li><li>Turner, J. G., Brozoski, T. J., Bauer, C. A., Parrish, J. L., Myers, K., Hughes, L. F., Caspary, D. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Gap+detection+deficits+in+rats+with+tinnitus:+a+potential+novel+screening+tool.">Gap detection deficits in rats with tinnitus: a potential novel screening tool.</a> Behav. Neurosci. 120, 188-195 (2006).</li><li>Turner, J., Larsen, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Relationship+between+noise+exposure+stimulus+properties+and+tinnitus+in+rats:+Results+of+a+12-month+longitudinal+study.">Relationship between noise exposure stimulus properties and tinnitus in rats: Results of a 12-month longitudinal study.</a> ARO. Abs. 594, (2012).</li><li>Turner, J. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Behavioral+measures+of+tinnitus+in+laboratory+animals.">Behavioral measures of tinnitus in laboratory animals.</a> Prog. Brain Res. 166, 147-156 (2007).</li><li>Engineer, N. D., Riley, J. R., Seale, J. D., Vrana, W. A., Shetake, J. A., Shetake, J. A., Sudanagunta, S. P., Borland, M. S., Kilgard, M. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Reversing+pathological+neural+activity+using+targeted+plasticity.">Reversing pathological neural activity using targeted plasticity.</a> Nature. 470, 101-104 (2011).</li></ol>

没有利益冲突的声明。

我们提出了一个基于在啮齿类动物的听力测量声惊吓反应，可用于确定行为听阈（听力图10）和听觉的幻象知觉，如主观耳鸣11前脉冲抑制价格便宜，而且易于制造安装。特别是后者的测量是在最近的几个报告8,12,13,14的焦点，并可以看作是一个先决条件，电调查这种疾病的神经元的机制。使用这种方法可以区分的动物没有发展一个主观的耳鸣声损伤和那些没有这样做一遍?...

a low cost setup for behavioral audiometry in rodents

在听觉动物研究的关键是听证会的基本参数的动物实验中所涉及的主题有准确的信息。这些参数可能是生理性的听觉通路的响应特性， 例如，通过脑干听力检查（BERA）。但是，这些方法允许对应于这些生理参数的听觉知觉，只是间接的和不确定的推断。为了评估的知觉的听力水平，行为的方法，必须使用。知觉的描述，在动物模型中使用的行为方式的一个潜在问题是，这些方法大多涉及某种行为测试的主题之前，可以学习的范例， 如动物可能要学会按杠杆响应于一个声音。由于这些学习范式改变自己的看法1,2，他们因此会影响人们获得任何结果约这些方法，因此必须谨慎解释。例外的是利用反射性反应的范例，因为在这里没有学习的范例之前进行感知测试。一个这样的反射反应是声惊吓反应（ASR），可高度重复性引起意想不到的响亮的声音在天真的动物。这ASR反过来可以影响取决于这个前刺激的感知性的声音之前：声音远高于听觉阈值将完全抑制的振幅的ASR;听起来接近阈值只会稍微抑制ASR。这种现象被称为预脉冲抑制（PPI）的3,4，和PPI的量上的ASR逐渐依赖于预脉冲的感知性。 PPI的ASR因此非常适合以确定行为的听力图天真，非受过训练的动物，以确定听力障碍，甚至以检测出可能在这些主观耳鸣知觉动物。在本文中，我们演示了如何使用这种方法，在啮齿类动物模型（参见文献5），长爪沙鼠（长爪沙鼠），这是一个正常人类的听觉范围内，众所周知模型物种的惊吓反应的研究（ 例如： 6）。

Watch this Scientific Journal Video about A Low Cost Setup for Behavioral Audiometry in Rodents at JoVE.com

A Low Cost Setup for Behavioral Audiometry in Rodents

听到的参数，如听阈，听力有障碍或幻象的看法（主观耳鸣）的行为确定为一种快速，廉价的方法。它使用预脉冲抑制声惊吓响应，并可以容易地实现在个人计算机中，使用一个可编程的AD / DA转换器和压电传感器。

在啮齿动物的行为测听的低成本设置

In auditory animal research it is crucial to have precise information about basic hearing parameters of the animal subjects that are involved in the ...

a-low-cost-setup-for-behavioral-audiometry-in-rodents

Research

JoVE Journal

Neuroscience

12.6K 次观看.  University of Erlangen-Nuremberg. 听到的参数，如听阈，听力有障碍或幻象的看法（主观耳鸣）的行为确定为一种快速，廉价的方法。它使用预脉冲抑制声惊吓响应，并可以容易地实现在个人计算机中，使用一个可编程的AD / DA转换器和压电传感器。

视频: A Low Cost Setup for Behavioral Audiometry in Rodents

A General Method for Evaluating Incubation of Sucrose Craving in Rats

For someone on a food-restricted diet, food craving in response to food-paired cues may serve as a key behavioral transition point between abstinence and relapse to food taking 1. Food craving conceptualized in this way is akin to drug craving in response to drug-paired cues. A rich literature has been developed around understanding the behavioral and neurobiological determinants of drug craving; we and others have been focusing recently on translating techniques from basic addiction research to better understand addiction-like behaviors related to food 2-4.
As done in previous studies of drug craving, we examine sucrose craving behavior by utilizing a rat model of relapse. In this model, rats self-administer either drug or food in sessions over several days. In a session, lever responding delivers the reward along with a tone+light stimulus. Craving behavior is then operationally defined as responding in a subsequent session where the reward is not available. Rats will reliably respond for the tone+light stimulus, likely due to its acquired conditioned reinforcing properties 5. This behavior is sometimes referred to as sucrose seeking or cue reactivity. In the present discussion we will use the term "sucrose craving" to subsume both of these constructs.
In the past decade, we have focused on how the length of time following reward self-administration influences reward craving. Interestingly, rats increase responding for the reward-paired cue over the course of several weeks of a period of forced-abstinence. This "incubation of craving" is observed in rats that have self-administered either food or drugs of abuse 4,6. This time-dependent increase in craving we have identified in the animal model may have great potential relevance to human drug and food addiction behaviors.
Here we present a protocol for assessing incubation of sucrose craving in rats. Variants of the procedure will be indicated where craving is assessed as responding for a discrete sucrose-paired cue following extinction of lever pressing within the sucrose self-administration context (Extinction without cues) or as responding for sucrose-paired cues in a general extinction context (Extinction with cues).

Responding for food or drug-paired cues increases over the course of a period of abstinence and this may relate to an increased susceptibility to relapse behaviors. Here we detail a procedure for evaluating this "incubation of craving" in rats that have self-administered sucrose.

评估大鼠蔗糖渴求育成的一个通用方法

For someone on a food-restricted diet, food craving in response to food-paired cues may serve as a key behavioral transition point between abstinence and ...

科研

神经科学

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 Lightweight, Headphones-based System for Manipulating Auditory Feedback in Songbirds

Experimental manipulations of sensory feedback during complex behavior have provided valuable insights into the computations underlying motor control and sensorimotor plasticity1. Consistent sensory perturbations result in compensatory changes in motor output, reflecting changes in feedforward motor control that reduce the experienced feedback error. By quantifying how different sensory feedback errors affect human behavior, prior studies have explored how visual signals are used to recalibrate arm movements2,3 and auditory feedback is used to modify speech production4-7. The strength of this approach rests on the ability to mimic naturalistic errors in behavior, allowing the experimenter to observe how experienced errors in production are used to recalibrate motor output.
Songbirds provide an excellent animal model for investigating the neural basis of sensorimotor control and plasticity8,9. The songbird brain provides a well-defined circuit in which the areas necessary for song learning are spatially separated from those required for song production, and neural recording and lesion studies have made significant advances in understanding how different brain areas contribute to vocal behavior9-12. However, the lack of a naturalistic error-correction paradigm - in which a known acoustic parameter is perturbed by the experimenter and then corrected by the songbird - has made it difficult to understand the computations underlying vocal learning or how different elements of the neural circuit contribute to the correction of vocal errors13.
The technique described here gives the experimenter precise control over auditory feedback errors in singing birds, allowing the introduction of arbitrary sensory errors that can be used to drive vocal learning. Online sound-processing equipment is used to introduce a known perturbation to the acoustics of song, and a miniaturized headphones apparatus is used to replace a songbird's natural auditory feedback with the perturbed signal in real time. We have used this paradigm to perturb the fundamental frequency (pitch) of auditory feedback in adult songbirds, providing the first demonstration that adult birds maintain vocal performance using error correction14. The present protocol can be used to implement a wide range of sensory feedback perturbations (including but not limited to pitch shifts) to investigate the computational and neurophysiological basis of vocal learning.

We describe the design and assembly of miniaturized headphones suitable for replacing a songbird&#x2019;s natural auditory feedback with a manipulated acoustic signal. Online sound processing hardware is used to manipulate song output, introduce real-time errors in auditory feedback via the headphones, and drive vocal motor learning.

一个轻量级的，耳机为基础的系统操作鸣禽听觉反馈

Experimental manipulations of sensory feedback during complex behavior have provided valuable insights into the computations underlying motor control and ...

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

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, including rats, hamsters, and mice, are active during the night and relatively inactive during the day. Many other behavioral and physiological measures also exhibit daily rhythms, but in rodents, running-wheel activity serves as a particularly reliable and convenient measure of the output of the master circadian clock, the suprachiasmatic nucleus (SCN) of the hypothalamus. In general, through a process called entrainment, the daily pattern of running-wheel activity will naturally align with the environmental light-dark cycle (LD cycle; e.g. 12 hr-light:12 hr-dark). However circadian rhythms are endogenously generated patterns in behavior that exhibit a ~24 hr period, and persist in constant darkness. Thus, in the absence of an LD cycle, the recording and analysis of running-wheel activity can be used to determine the subjective time-of-day. Because these rhythms are directed by the circadian clock the subjective time-of-day is referred to as the circadian time (CT). In contrast, when an LD cycle is present, the time-of-day that is determined by the environmental LD cycle is called the zeitgeber time (ZT).
Although circadian rhythms in running-wheel activity are typically linked to the SCN clock6-8, circadian oscillators in many other regions of the brain and body9-14 could also be involved in the regulation of daily activity rhythms. For instance, daily rhythms in food-anticipatory activity do not require the SCN15,16 and instead, are correlated with changes in the activity of extra-SCN oscillators17-20. Thus, running-wheel activity recordings can provide important behavioral information not only about the output of the master SCN clock, but also on the activity of extra-SCN oscillators. Below we describe the equipment and methods used to record, analyze and display circadian locomotor activity rhythms in laboratory rodents.

Circadian rhythms in voluntary wheel-running activity in mammals are tightly coupled to the molecular oscillations of a master clock in the brain. As such, these daily rhythms in behavior can be used to study the influence of genetic, pharmacological, and environmental factors on the functioning of this circadian clock.

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, ...

A Low-cost Method for Analyzing Seizure-like Activity and Movement in Drosophila

Video tracking systems have been used widely to analyze Drosophila melanogaster movement and detect various abnormalities in locomotive behavior. While these systems can provide a wealth of behavioral information, the cost and complexity of these systems can be prohibitive for many labs. We have developed a low-cost assay for measuring locomotive behavior and seizure movement in D. melanogaster. The system uses a web-cam to capture images that can be processed using a combination of inexpensive and free software to track the distance moved, the average velocity of movement and the duration of movement during a specified time-span. To demonstrate the utility of this system, we examined a group of D. melanogaster mutants, the Bang-sensitive (BS) paralytics, which are 3-10 times more susceptible to seizure-like activity (SLA) than wild type flies. Using this novel system, we were able to detect that the BS mutant bang senseless (bss) exhibits lower levels of exploratory locomotion in a novel environment than wild type flies. In addition, the system was used to identify that the drug metformin, which is commonly used to treat type II diabetes, reduces the intensity of SLA in the BS mutants.

A Low-cost Method for Analyzing Seizure-like Activity and Movement in Drosophila

Using a webcam and a combination of free and inexpensive software programs, locomotive patterns in Drosophila melanogaster can be analyzed to detect differences in the speed, distance, and time of various motor activities.

在分析癫痫样活动和运动的一种低成本方法<em&gt;果蝇</em

Video tracking systems have been used widely to analyze Drosophila melanogaster movement and detect various abnormalities in locomotive behavior. While ...

A Human-machine-interface Integrating Low-cost Sensors with a Neuromuscular Electrical Stimulation System for Post-stroke Balance Rehabilitation

A stroke is caused when an artery carrying blood from heart to an area in the brain bursts or a clot obstructs the blood flow to brain thereby preventing delivery of oxygen and nutrients. About half of the stroke survivors are left with some degree of disability. Innovative methodologies for restorative neurorehabilitation are urgently required to reduce long-term disability. The ability of the nervous system to reorganize its structure, function and connections as a response to intrinsic or extrinsic stimuli is called neuroplasticity. Neuroplasticity is involved in post-stroke functional disturbances, but also in rehabilitation. Beneficial neuroplastic changes may be facilitated with non-invasive electrotherapy, such as neuromuscular electrical stimulation (NMES) and sensory electrical stimulation (SES). NMES involves coordinated electrical stimulation of motor nerves and muscles to activate them with continuous short pulses of electrical current while SES involves stimulation of sensory nerves with electrical current resulting in sensations that vary from barely perceivable to highly unpleasant. Here, active cortical participation in rehabilitation procedures may be facilitated by driving the non-invasive electrotherapy with biosignals (electromyogram (EMG), electroencephalogram (EEG), electrooculogram (EOG)) that represent simultaneous active perception and volitional effort. To achieve this in a resource-poor setting, e.g., in low- and middle-income countries, we present a low-cost human-machine-interface (HMI) by leveraging recent advances in off-the-shelf video game sensor technology. In this paper, we discuss the open-source software interface that integrates low-cost off-the-shelf sensors for visual-auditory biofeedback with non-invasive electrotherapy to assist postural control during balance rehabilitation. We demonstrate the proof-of-concept on healthy volunteers.

A novel low-cost human-machine interface for interactive post-stroke balance rehabilitation system is presented in this article. The system integrates off-the-shelf low-cost sensors towards volitionally driven electrotherapy paradigm. The proof-of-concept software interface is demonstrated on healthy volunteers.

人机界面集成的低成本传感器，具有神经肌肉电刺激系统中风后康复平衡

A stroke is caused when an artery carrying blood from heart to an area in the brain bursts or a clot obstructs the blood flow to brain thereby preventing ...

High-density Electroencephalographic Acquisition in a Rodent Model Using Low-cost and Open-source Resources

Advanced electroencephalographic analysis techniques requiring high spatial resolution, including electrical source imaging and measures of network connectivity, are applicable to an expanding variety of questions in neuroscience. Performing these kinds of analyses in a rodent model requires higher electrode density than traditional screw electrodes can accomplish. While higher-density electroencephalographic montages for rodents exist, they are of limited availability to most researchers, are not robust enough for repeated experiments over an extended period of time, or are limited to use in anesthetized rodents.1-3 A proposed low-cost method for constructing a durable, high-count, transcranial electrode array, consisting of bilaterally implantable headpieces is investigated as a means to perform advanced electroencephalogram analyses in mice or rats.

Procedures for headpiece fabrication and surgical implantation necessary to produce high signal to noise, low-impedance electroencephalographic and electromyographic signals are presented. While the methodology is useful in both rats and mice, this manuscript focuses on the more challenging implementation for the smaller mouse skull. Freely moving mice are only tethered to cables via a common adapter during recording. One version of this electrode system that includes 26 electroencephalographic channels and 4 electromyographic channels is described below.

Instructions for the low-cost construction and surgical implantation of a chronic transcranial high-density electroencephalographic montage into mice are provided. Signal recording, extraction, and processing techniques are also described.

高密度的脑电图采集在啮齿动物模型使用低成本的开源资源

Advanced electroencephalographic analysis techniques requiring high spatial resolution, including electrical source imaging and measures of network ...

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.

间刺激间隔对啮齿类动物时间处理评估的力量

Temporal processing deficits have been implicated as a potential elemental dimension of higher-level cognitive processes, commonly observed in ...

A Hydrophobic Tissue Clearing Method for Rat Brain Tissue

Hydrophobic tissue clearing methods are easily adjustable, fast, and low-cost procedures that allows for the study of a molecule of interest in unaltered tissue samples. Traditional immunolabeling procedures require cutting the sample into thin sections, which restricts the ability to label and examine intact structures. However, if brain tissue can remain intact during processing, structures and circuits can remain intact for the analysis. Previously established clearing methods take significant time to completely clear the tissue, and the harsh chemicals can often damage sensitive antibodies. The iDISCO method quickly and completely clears tissue, is compatible with many antibodies, and requires no special lab equipment. This technique was initially validated for the use in mice tissue, but the current protocol adapts this method to image hemispheres of control and transgenic rat brains. In addition to this, the present protocol also makes several adjustments to preexisting protocol to provide clearer images with less background staining. Antibodies for Iba-1 and tyrosine hydroxylase were validated in the HIV-1 transgenic rat and in F344/N control rats using the present hydrophobic tissue clearing method. The brain is an interwoven network, where structures work together more often than separately of one another. Analyzing the brain as a whole system as opposed to a combination of individual pieces is the greatest benefit of this whole brain clearing method.

Here we present a hydrophobic tissue clearing method that allows for the viewing of target molecules as part of intact brain structures. This technique has now been validated for F344/N control and HIV-1 transgenic rats of both sexes.

鼠脑组织疏水组织清除方法

Hydrophobic tissue clearing methods are easily adjustable, fast, and low-cost procedures that allows for the study of a molecule of interest in unaltered ...

A Rat Model of EcoHIV Brain Infection

It has been well studied that the EcoHIV infected mouse model is of significant utility in investigating HIV associated neurological complications. Establishment of the EcoHIV infected rat model for studies of drug abuse and neurocognitive disorders, would be beneficial in the study of neuroHIV and HIV-1 associated neurocognitive disorders (HAND). In the present study, we demonstrate the successful creation of a rat model of active HIV infection using chimeric HIV (EcoHIV). First, the lentiviral construct of EcoHIV was packaged in cultured 293 FT cells for 48 hours. Then, the conditional medium was concentrated and titered. Next, we performed bilateral stereotaxic injections of the EcoHIV-EGFP into F344/N rat brain tissue. One week after infection, EGFP fluorescence signals were detected in the infected brain tissue, indicating that EcoHIV successfully induces an active HIV infection in rats. In addition, immunostaining for the microglial cell marker, Iba1, was performed. The results indicated that microglia were the predominant cell type harboring EcoHIV. Furthermore, EcoHIV rats exhibited alterations in temporal processing, a potential underlying neurobehavioral mechanism of HAND as well as synaptic dysfunction eight weeks after infection. Collectively, the present study extends the EcoHIV model of HIV-1 infection to the rat offering a valuable biological system to study HIV-1 viral reservoirs in the brain as well as HAND and associated comorbidities such as drug abuse.

Here, we present a protocol to establish a new rat model of active HIV infection using chimeric HIV (EcoHIV), which is critical for enhancing our understanding of HIV-1 viral reservoirs in the brain and offering a system to study HIV-associated neurocognitive disorders and associated comorbidities (i.e., drug abuse).

生态HIV脑感染的老鼠模型

It has been well studied that the EcoHIV infected mouse model is of significant utility in investigating HIV associated neurological complications. ...