（; BH史密斯CO-PIĴTrumble PI）和亚利桑那州立大学的这项研究得到了美国国立卫生研究院从NCRR（R01 RR014166到BHS），美国国立卫生研究院NIDCD（R01 DC011422 BHS的合作，PI），美国农业部门的资金支持。资金的工作坊，以训练学生（ 图4的数据）是由智利科学基金会提供的。

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The authors have no conflicts of interest to disclose.

该协议提出了使用每次操作空调2可靠的方法。这是两个的几种方法已被成功地应用27，21。我们采用方法2，使用市盈率，因为它是一贯可靠的，甚至可以跨越不同的实验者所有的实验研究。 相同的基本过程已经适应了许多不同种类的研究，蜜蜂，包括使用不同的条件性刺激和不同的行为反射的。它也被链接到的差异的遗传基础的调查，在学习28,29，在大脑13,14,30嗅觉感知和记忆的生理相关因素，以及行为15,16,31的调节和分子遗传学基础。由于嗅觉学习蜜蜂生态相关的先进知识，这在100多年前开始与卡尔·冯·弗里希的首批研究<suP&gt; 31，每结果可以很容易地连接到一个群体的生存需要。最近，已经适于农业研究，以农药和环境毒素17的亚致死行为效应。 基本程序是在于其可应用于研究其他物种的问题，以及强大。两个蛾类， 烟草天蛾和海灰翅夜蛾 ，已经使用的PER的研究来探讨气味的识别和各物种的嗅觉学习能力32,33的神经基础。每实验果蝇提供了许多见解参与chemosensation和学习34的分子信号级联反应。每近来被用来研究习惯的跳蚤（Rhodnius蝽 ）35，一个重要的病媒。 在一般情况下，该过程是健壮的变化的方法;采用的Different方法可能会产生治疗组之间相同的相对差异。尽管相对简单的过程，有几个问题可以在时间防止蜜蜂学习的气味蔗糖的关联。下面的主题是可能改变的协议，一些在市盈率空调可能出现的一些常见问题。 数量上的考虑和试验类型的调理协议学习任何协议都需要蜜蜂暴露了一些在采集阶段的试验。这个数目在很大程度上取决于任务的难度。蜜蜂能学会审单后，一个简单的任务，但他们至少需要三个试验诱发长期记忆的形成。正如所料，蜜蜂需要显著更多的试验，以充分了解一项艰巨的任务。通常，存在试验超出该蜂不再显著提高的最大数目IR的表现。该最大值取决于特​​定任务时，气味的类型和浓度，以及在蔗糖浓度。 当使用多于一种类型的试验用不同的气味，呈现出不同的试验类型中的伪随机序列，以保持蜜蜂从记忆的气味演示了一个简单序列，而不是学习的气味36之间的差异。在这些伪随机序列，必须有相等数量的试验中对每个气味类型的。此外，一种香型审判的可能性正在前面和后面相同的气味，或其他任何异味，必须等于所有的气味。二气味 - 甲乙 - 使用下面的顺序：ABBABAAB。在八项试验每个气味呈现四倍。串连相同的序列，以达到试验每个气味的期望数目。 无论是实验设计的，也有必须保持一些参数常数以优化学习。 CS和美国之间的重叠是有效的调节至关重要。该试验间隔（ 图1，表1）需要是常数并且被优化，因为不规则或太短或太长试验间隔可以显著影响调理方案1的效果。 在行为测试试验中的重要性及设计注意事项在一个实验中，当CS和美国一起呈现的“采集阶段”记录的反应，可能是有用的。不过，要注意的气味呈现条件不同于一种试验到另一个。在增强试验中，气味接着蔗糖，提出结合视觉刺激（装置移动用于输送蔗糖液滴）可以影响蜜蜂的响应。和蜜蜂只有3秒s到显示的响应（4秒气味刺激减去与蔗糖陈述所述一个秒的重叠， 图1）。如果实验涉及的蜜蜂学习区分2气味（ 如钢筋和无钢筋），未增强的气味呈现交替试验时不会奖励呈现的视觉刺激和蜜蜂有整整四秒的时间。因此，应对这两个气味是不是采集过程完全可比。对于任何调理协议，不收购曲线1只依赖。为了更好地确定哪些蜜蜂了解到，推出了一系列的非增强型测试试验，在此期间，没有气味增强，从而确保所有刺激试验时间相同的条件下进行。 根据实验的具体目的，测试可以由经调节的气味或一系列的试验与空调外径的单个试验的试验或或空调，新气味的组合。在空调异味的单个测试试验提供的蜜蜂是否还记得空调异味的简单分析。然而，响应于所述第一测试试验可能反映了蜜蜂动机水平以及对调节后的气味的召回。的一系列测试的实验中，无论是串联的空调气味或一系列的条件和一个或多个新的气味的单一测试的消光的实验中，也可以被用来评估存储器。该系列灭绝试验可以评估的蜜蜂多么强烈还记得空调协会。该召回强更多扑灭条件反应所需的试验。一系列的条件和新的气味单一测试也可以评估的特异性的调节气味的蜜蜂的记忆。 此外，还必须进行调理，同时测试对照和处理组在相同的时间点。对比例空调要保持较长时期的蜜蜂后不久，荷兰国际集团蜜蜂的表现是因为接触到蔗糖美国的蜜蜂喂以饱食的问题。例如，长的时间间隔之后的降低性能可能是由于存储器腐烂，或者它可能是由于由未增强曝光感应与美国变化的激励状态和/或学习，使得结果模棱两可。因此，治疗组的性能应始终判断相对于对照组的空调，并在同一时间点进行测试。 臭气浓度和完整性有几种方法，其中所述气味（CS）的浓度和完整性可能会受到影响。最普遍的问题与气味递送的气味墨盒耗尽从过度使用（ 图2）。更换墨盒10-12用途（每2或3项试验为5蜜蜂组），以避免这一缺陷。同样重要的是使用新鲜的墨盒测试试验，因为用过的碳粉盒可耗尽的差异，因此目前不平等的刺激气味。另一个常见的​​问题是气味盒污染由于使用盒，一个以上的气味不完全清洁它。脏或被污染的空气流动也无意引入额外的嗅觉刺激（活性炭过滤器可以防止后台污染）。 PER调理与生理反应的气味测量耦合时，这是特别有问题的。漏气味墨盒出现问题，因为蜜蜂暴露于气味试验开始前，从而降低了刺激气味的显着性。在空气供给对气味输送宽松可导致很少或没有气味输送当阀打开时，人为地降低了蜜蜂响应于预定的气味。 蔗糖溶液和非条件刺激（奖励） 在A安装，浓缩，并作为美国的蔗糖溶液的完整性是对实验的成功是至关重要的。调节是作为美国37的量和蔗糖水溶液浓度的函数。在方法2中使用的微米注射器允许非常精确（下降到0.2微升）控制美国的交付，我们建议您使用它们的描述这两种方法。用牙签（方法1）是足够的情况下，其中昂贵的注射器不能用，比如，在训练大群学生，在外地工作，或以较低的预算。认真执行方法1是罚款，只要定时保持着定期更换牙签，避免蔗糖的积累（和交付的浓度）。但是，使用牙签它更难以准确地控制和估计量和美国的幅度传递，其用于调节的研究1重要。蔗糖的所需浓度提供足够的奖励，以保持主动地学习了空调协会可以根据操作任务的难度和蜜蜂的内部状态或一年时间的蜜蜂。更困难的任务需要蜜蜂的成功学习任务更高的蔗糖溶液。即使在5℃的蔗糖溶液模具可累积，这将危及该解决方案的完整性，在实验过程中影响了蜜蜂的健康和感知的奖励。这是最好的替代解决方案，每隔几天。 精密，定时，以及CS和美国交付的一致性关于正确实施每道工序，或与此有关的任何调理过程中，最关键的问题涉及精度，时间和CS的一致性和美国交付（ 图1）。调查谁是新的程序往往是不精确的约交付一个或两个刺激。三军情报局的未能以允许重叠的CS和US导致空调性能不佳。 PLC的自动的可听信号，用于实验者气味输送发病后交付蔗糖3秒。调查人员应持有的蔗糖/水滴接近蜜蜂的信号后，迅速交货。这些程序帮助夹带一致的ISI。通过调节舞台上放置一个秒表，以及允许在审讯方便的时间作为安置之前和分娩的刺激后，监测蜜蜂一次在舞台上。这样的ITI的可以是相对一致的并且整个过程可以在受控的速度下运行。 ITI的过短，例如小于1分钟，或太长会导致空调性能差1。 对蜜蜂业绩的季节性，环境和语境效果蜜蜂的表现可以用它的周围被显著影响之前和调节过程中。通常情况下，来与四季变化的温度和食物供应的波动会改变蜜蜂的学习动机。当鲜花盛开，蜜蜂的学习动机在实验室条件的关联降低38。当殖民地强调 - 从极端温度，食物短缺，或疾病有关的压力 - 蜜蜂&#39;将显示在他们的学习表现下降。蜜蜂不停地在飞行的房间可以好好学习了一段时间，但有太多的疾病的压力和老化随着时间的推移他们的学习表现。调节过程中的环境还可以降低蜜蜂的表现。任何多余的气味，运动和其他刺激可以从实验的刺激分散的蜂。为了避免这个问题，保持了相当一致，简化的视觉环境。 基因型和经验影响每场演出 &lt;P舱=“jove_content”&gt;蜜蜂工人可以在性能明显不同于任何调理过程由于任务专业化，基因型，或其他环境因素20。因此，标准化，尽可能地，用在一个实验中，以动物的类型，以减少个体间差异是很重要的。在领导的开放式交配女王，这意味着她交配与许多不同的无人驾驶飞机的殖民地，工人将不同的父本基因型。遗传背景可导致感官反应39戏剧性的差异和学习表现29。使用菌落率领皇后工具性从单一的无人机28人工授精精子减少这种个体间差异。 以上所述的协议，包括用于从巢入口收集蜜蜂工人的方法。然而，这些蜜蜂彼此在考虑年龄或行为的任务不同。他们可能是年轻（inexperienCED）以上（比较有经验）觅食。他们可能是年轻的蜜蜂使他们的第一个方向飞行。或者，他们可能是守卫的蜜蜂。为了减少变异，标志着具有快干磁漆或标识标签蜜蜂要么因为他们脱颖而出，成为成年人（控制年龄）和/或他们开始觅食（以控制经验）。然后，几天后，收集标记的蜜蜂进行调理。在从事护理工作者的行为可以从蜂巢中的帧进行收集。护理人员可当他们将他们的头到育雏细胞喂养和照顾幼虫内被验明正身。 用处女皇后或无人机每调理除了 ​​工人蜜蜂，处女蜜蜂蜂王和雄蜂可以很容易调节的市盈率过程的研究旨在发展蜜蜂在不同的学习表现28的遗传线路。处女蜂王应尽快日之后收集安永从化蛹出现，并直接放入约束的线束没有麻醉。从殖民地内育雏梳收集年轻，不成熟的无人机一般都不会主动地学习。之后，他们已经开始交配飞行，无人机很容易了解每任务28。因为他们从配合航班返回，并维持隔夜在一个小笼子中用于饲养女王的殖民地，他们应该被收集。不要试图调节的前一天，以利用它们;他们没有在一夜之间吊带存活良好。一对夫妇的前调理小时的无人机可以从笼子里被收集并放入约束的线束没有麻醉。 结论此PER程序，在方法的方式，相当于一个起点设计PER实验。最PER协议将要求概述以某种方式来改变的程序来实现的实验和accommodat的具体目标E倍数治疗组。这是很容易实现。然而，正确的实施需要对细节的关注和实践。一旦掌握了它可以是一个功能强大的程序添加为研究工具用于不同的昆虫种类数的基础和应用研究项目。

许多昆虫了解生态相关的刺激，然后他们改变他们的行为反应的那些刺激，以适应其环境中的新的预测关系。几种不同的机制可以这背后行为可塑性（ 例如 ，非关联，关联/巴甫洛夫和操作性/器乐1）。这些类型的可塑性的不同之处是如何刺激或行为都与重要事件，如食品的出现，伴侣，或危险有关。理解这些形式的可塑性是用于基础研究非常重要成神经系统如何变化来编码新的记忆2。这也是理解昆虫是疾病的重要媒介（ 如 ，采采蝇和蚊子）和昆虫是农业上重要的适应性行为重要的是，无论是在作物生产（蜜蜂）或害虫。 学习行为可塑性任何阿尼姆人需要实验控制的电平超过了一些变量是不能达到的，在字段1。它要求能更加控制的条件下被使用，但它仍然是相关与在自然条件下的行为健壮调理协议的发展。蜜蜂（ 蜜蜂 ）是如何开发的协议，用于执行行为可塑性3,4对照分析一个很好的典范。长鼻扩展响应（PER）的蜜蜂是一种自然的行为反射在蜜蜂扩展其长鼻回应触角刺激糖溶液。在正常的觅食行为，每当蜜蜂发现蜜源的花出现。幸运的是，蜜蜂很容易表现在实验室这个简单而容易量化的行为。这使得有可能研究，在受控制的环境，影响这种生态相关的行为5的机制。每CAÑ ​​还可以调理协议中用于研究不同的处理条件，其目的是为了揭示背后的可塑性6的行为和神经机制的刺激下，知觉，学习与记忆。 由于由桑原7所述第一研究中，每调理已广泛用于揭示非结合，缔合和背后行为可塑性在蜜蜂8操作性的机制。这些机制中的那些相同的自由飞行蜜蜂9的研究显示。不同于自由飞翔蜜蜂的研究，每调理协议可以与电10,11或脑活细胞荧光成像12-14。此外，PER协议允许经由药物或分子遗传学的治疗实验操作的神经通路的测试有关网络的特定组件，例如neur的作用假说omodulators 15,16。市盈率协议也提供了评估的环境条件，以及对健康的毒素和蜜蜂17的觅食效率的亚致死效应的重要途径。 此过程说明了两个并行的气味传递方法。方法1是一个版本的气味和非条件刺激（蔗糖）交付，提供了一个价格低廉，技术简单的方法呈现的气味和蔗糖的奖励。这种方法是很好的基础训练，当自动化是不可能的。这是介绍这种技术到教室或教学实验室的极佳方式。在调理涉及更困难的任务和气味的感知，学习，或存储器的耦合生理评估实验，这是非常重要的，以精确地和精确地调节刺激的发作，持续时间递送定时。对于最可靠的刺激传递，使用自动化的气味交付的手段ND奖励交货的精确方法。方法2采用自动化的气味传递和更精确的蔗糖交付。它在技术上是更复杂的，并且需要更多的初始设置比方法1，但它显著增加用于调节刺激的定时和数量的一致性，应尽可能使用。

<table border="0" cellpadding="0" cellspacing="0" width="1278">
	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:371px;">Name of Material/ Equipment</td>
			<td style="width:203px;">Company</td>
			<td style="width:119px;">Catalog Number</td>
			<td style="width:587px;">Comments/Description</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:371px;">Sucrose</td>
			<td style="width:203px;">Sigma-Aldrich</td>
			<td style="width:119px;">S9378-1KG</td>
			<td style="width:587px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:371px;">Odorant compounds (for example):</td>
			<td style="width:203px;"></td>
			<td style="width:119px;"></td>
			<td rowspan="6" style="width:587px;">For additional examples of odorants, see any of the papers on olfactory processing from the Smith lab</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:371px;">1-hexanol</td>
			<td style="width:203px;">Sigma-Aldrich</td>
			<td style="width:119px;">471402-100ML</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:371px;">2-octanone</td>
			<td style="width:203px;">Sigma-Aldrich</td>
			<td style="width:119px;">W280208-800G-K</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:371px;">heptanol</td>
			<td style="width:203px;">Sigma-Aldrich</td>
			<td style="width:119px;">51778-5ML</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:371px;">gerianol</td>
			<td style="width:203px;">Sigma-Aldrich</td>
			<td style="width:119px;">163333-25G</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:371px;">nonanal</td>
			<td style="width:203px;">Sigma-Aldrich</td>
			<td style="width:119px;">131210-100ML</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:371px;">Hexane</td>
			<td style="width:203px;">Sigma-Aldrich</td>
			<td style="width:119px;">296090-1L</td>
			<td style="width:587px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:371px;">Heavy Mineral Oil</td>
			<td style="width:203px;">Sigma-Aldrich</td>
			<td style="width:119px;">330760-1L</td>
			<td style="width:587px;">Make sure it&#8217;s odorless</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:371px;">Ethanol</td>
			<td style="width:203px;">Sigma-Aldrich</td>
			<td style="width:119px;">459836-1L</td>
			<td style="width:587px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:371px;">Scintillation vials</td>
			<td style="width:203px;">Sigma-Aldrich</td>
			<td style="width:119px;">Z190535-1PAK</td>
			<td style="width:587px;">Use a small drill bit to bore a small hole in the cap of the vials&#160;</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:371px;">Bee harness</td>
			<td style="width:203px;"></td>
			<td style="width:119px;"></td>
			<td style="width:587px;">Custom-made from 0.9 cm diameter plastic soda straws or hard plastic/metal tubing</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:371px;">Duct tape</td>
			<td style="width:203px;"></td>
			<td style="width:119px;"></td>
			<td style="width:587px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:371px;">Kimwipes</td>
			<td style="width:203px;">Sigma-Aldrich</td>
			<td style="width:119px;">Z188956-1PAK</td>
			<td style="width:587px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:371px;">Wash bottles</td>
			<td style="width:203px;">Sigma-Aldrich</td>
			<td style="width:119px;">Z560847-3EA</td>
			<td style="width:587px;">For the 70% ethanol&#160;</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:371px;">Small fan in mountable housing</td>
			<td style="width:203px;"></td>
			<td style="width:119px;"></td>
			<td style="width:587px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:371px;">Dryer tubing</td>
			<td style="width:203px;"></td>
			<td style="width:119px;"></td>
			<td style="width:587px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:371px;"></td>
			<td style="width:203px;"></td>
			<td style="width:119px;"></td>
			<td style="width:587px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:371px;"></td>
			<td style="width:203px;"></td>
			<td style="width:119px;"></td>
			<td style="width:587px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:371px;"></td>
			<td style="width:203px;"></td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:371px;">FOR METHOD 1 ONLY</td>
			<td style="width:203px;"></td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:371px;">Name</td>
			<td style="width:203px;">Company</td>
			<td style="width:119px;">Catalog Number</td>
			<td style="width:587px;">Comments</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:371px;">20 ml disposable plastic syringes</td>
			<td style="width:203px;">Cole-Parmer</td>
			<td style="width:119px;">WU-07945-18</td>
			<td style="width:587px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:371px;">15 mm filter paper circles</td>
			<td style="width:203px;">Sigma-Aldrich</td>
			<td style="width:119px;">Z274844-1PAK</td>
			<td style="width:587px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:371px;">Pushpins</td>
			<td style="width:203px;"></td>
			<td style="width:119px;"></td>
			<td style="width:587px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:371px;">Toothpicks</td>
			<td style="width:203px;"></td>
			<td style="width:119px;"></td>
			<td style="width:587px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:371px;"></td>
			<td style="width:203px;"></td>
			<td style="width:119px;"></td>
			<td style="width:587px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:371px;"></td>
			<td style="width:203px;"></td>
			<td style="width:119px;"></td>
			<td style="width:587px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:371px;"></td>
			<td style="width:203px;"></td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:371px;">FOR METHOD 2 ONLY</td>
			<td style="width:203px;"></td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:371px;">Name</td>
			<td style="width:203px;">Company</td>
			<td style="width:119px;">Catalog Number</td>
			<td style="width:587px;">Comments</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:371px;">Gilmont Micrometer syringe, 0.2 mL</td>
			<td style="width:203px;">Cole-Parmer</td>
			<td style="width:119px;">EW-07840-00</td>
			<td style="width:587px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:371px;">Gilmont micrometer syringe tip</td>
			<td style="width:203px;">Cole-Parmer</td>
			<td style="width:119px;">EW-07841-00</td>
			<td style="width:587px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:371px;">26G 3/8&#8221; Leur hub hypodermic needles</td>
			<td style="width:203px;">Fisher Scientific</td>
			<td style="width:119px;">14-826-10</td>
			<td style="width:587px;"></td>
		</tr>
		<tr height="63">
			<td height="63" style="height:63px;width:371px;">1cc tuberculin syringes (plastic/glass)</td>
			<td style="width:203px;">Sigma-Aldrich</td>
			<td style="width:119px;">Z181641-1EA&#160;&#160; OR &#160;&#160;Z192090-200EA</td>
			<td style="width:587px;">glass tuberculin syringes are available, but plastic syringes are much less expensive and will work well for a limited number of uses</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Small rubber/silicone restrictors</td>
			<td>Cole-Parmer</td>
			<td>EW-95702-02</td>
			<td>Made from 4.8 mm outer diameter silicone tubing or the rubber tips of the 1 cc syringe plungers</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:371px;">Parafilm</td>
			<td style="width:203px;">Sigma-Aldrich</td>
			<td style="width:119px;">P7793-1EA</td>
			<td style="width:587px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:371px;">75 X 100 mm filter paper</td>
			<td style="width:203px;">Sigma-Aldrich</td>
			<td style="width:119px;">Z695106-500EA</td>
			<td style="width:587px;"></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:371px;">Direct Logic 05 Programmable Logic Controller</td>
			<td style="width:203px;">Koyo Electronics Industries Co, Ltd</td>
			<td style="width:119px;"></td>
			<td style="width:587px;">http://www.koyoele.co.jp/&#160;&#160;&#160;&#160;&#160;&#160;&#160; OR&#160;&#160;&#160;&#160;&#160;&#160; english/support/dlc.html#plc</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:371px;">1 mm, 4mm &#38; 6 mm inner diameter PVC or silicone tubing&#160;</td>
			<td style="width:203px;">Cole-Parmer</td>
			<td style="width:119px;">Various</td>
			<td style="width:587px;">Cole-Parmer has a wide selection of suitable tubing</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:371px;">Polypropylene connectors &#38; leur fittings</td>
			<td style="width:203px;">Cole-Parmer</td>
			<td style="width:119px;">Various</td>
			<td style="width:587px;">Cole-Parmer has a wide selection of connectors and fittings for many tube sizes</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:371px;">65-mm Correlated Flowmeter</td>
			<td style="width:203px;">Cole-Parmer</td>
			<td style="width:119px;">EW-03216-08</td>
			<td style="width:587px;">Aluminum with Glass float; For Liquids and Gases, With Valve</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:371px;">&#160;&#160;&#160;&#160; OR&#160;</td>
			<td style="width:203px;"></td>
			<td style="width:119px;"></td>
			<td style="width:587px;"></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:371px;">Tetra Whisper 300 (Tetratek DW96-2) Aquarium Air Pump</td>
			<td style="width:203px;">Aquacave</td>
			<td style="width:119px;">AE-TETRA-300</td>
			<td style="width:587px;">www.aquacave.com</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:371px;">LIF series Solenoid Valves for .042 &#34; ID Tubing, Configuration &#34;E&#34;</td>
			<td style="width:203px;">The Lee Company</td>
			<td style="width:119px;">LFAA1200118H</td>
			<td style="width:587px;">Neoprene, 430 SS, 302 SS, 280mWatts&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160;&#160; http://www.theleeco.com/LEEWEB2.NSF</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:371px;">PC-Board 12VDC 70dB Piezo Buzzer</td>
			<td style="width:203px;">RadioShack</td>
			<td style="width:119px;">273-074</td>
			<td style="width:587px;">www.radioshack.com</td>
		</tr>
	</tbody>
</table>

a proboscis extension response protocol for investigating behavioral plasticity in insects: application to basic, biomedical, and agricultural research

昆虫修改他们的反应与生存（ 如食物，配偶，威胁）重要事件相关联的刺激体验刺激通过。还有，通过该昆虫学会凸协会和它们涉及这些事件的几个行为机制。重要的是要理解这个行为可塑性的，旨在帮助对昆虫有利于农业的程序是很重要的。这种理解，也可用于发现解决由昆虫作为传病媒介和害虫创建生物医学和农业问题。长鼻扩展响应（PER）的调理方案，开发了蜜蜂（ 蜜蜂 ）在50年前，研究他们是如何看待和了解花香气味，而信号的花蜜和花粉资源的殖民地需要生存。在每次操作提供了一个强大和易于运用框架学习几种不同的生态有关机甲行为可塑性的共享机制。它是为使用容易适应与其他几种昆虫物种和其他行为的反射。这些协议可以很容易地用在各种装置，用于通过电生理或生物成像监测神经活性在中枢神经系统一起使用，或者用于操纵靶向神经调节途径。它是一个强大的检测快速检测造成的环境压力，毒素或农药的行为亚致死效应。 我们将展示如何在每个协议是简单的实现使用两个程序。一种适合作为实验室练习为学生或对实验性治疗的效果的快速检测。另一个提供更彻底的控制变量，它是行为调节的研究很重要。我们显示如何若干措施的行为反应，从二进制是/否到像延迟和长鼻延伸的持续时间更连续变量可以用来测试hypothese第而且，我们讨论了一些陷阱，当他们使用的程序的第一次，研究人员经常遇到的问题。

我们提出了使用上述的PER的协议的两个示例。第一个例子利用方法2，研究蜜蜂如何感知不同气味的分子相似的功能，气味作为CS 5,24,25。第二种是使用方法1，有些必须开始时，每调节的实验用在研究实验室应采取的预防措施的一个例子。 研究像这样的已被用来形容结合蜜蜂和蛾类的嗅觉“感性空间”与生物成像13或电分析26。蜜蜂组（n = 20）进行调节的气味癸醛与蔗糖联想加固超过12前锋配对空调试验（ 图3A）。大约10％的蜜蜂在一审判决后，这是正常的反应&#39;自发&#39;。在那之后的responde的百分比ð增长在未来几年的试验，直到蜜蜂100％的受访者在第五审判，以后每试。几项研究已经表明，这种增加的响应是特定于向前的配对状态（ 图1）相对于几个重要的控制程序3。采集阶段后，每个蜜蜂进行了一系列的非增强型测试试验（ 图3B），其中涉及曝光的气味CS和该系统从CS中的分子结构不同其他一些增味剂。蜜蜂反应最强烈的CS。他们对其他气味物质的反应下降，作为结构的系统性变化的函数，用最不喜欢CS的加臭剂的最低响应。 在图4中，从最近的研讨会，让学生了解到，第一次的结果，如何调理用图示方法1蜜蜂。学生冷杉<html>吨空调蜜蜂通过正向配对methycyclohexanone（MCH）和辛醇（OCT）与蔗糖加固（ 图4A）。 ％的反应是低的第一次试验和增加对后续试验，表明该蜂学到的气味蔗糖关联。有在不同程度的蜜蜂学会在不同的学生群体的条件相关。在我们的经验中，这些种类的差异可能是由于在作为CS中的异味，在方法（方法1）的不精确的元素，或向实验者的训练水平。在第三种情况下，性能迅速成为与实验者“增加经验更一致。调理后，将各蜜蜂用每气味测试两次。在已被空调与华侨城或妇幼保健正向配对蜜蜂群体，反应一如预期，强劲的空调异味（ 图4B）。 <html> ENT“&gt;在经历着配对至十月蜜蜂的第二组空调在平行于另一组中的气味华侨城向后配对蔗糖。倒向配对是几种类型的控制程序来演示之一，增加反应是特定转发配对，而不是由于其它进程，诸如从致敏非特异性觉醒。为了向后配对，蔗糖美国和气味CS从向后配对呈现相反的顺序是什么， 如图1所示。采集数据是未示出，因为CR不能得分给定的气味CS上介绍的蔗糖之后的呈现。正如所预测的缔调理，响应水平华侨城分别向前配对组相对于所述向后组（ 图4C）中的高。 7fig1highres.jpg“WIDTH =”500“/&gt; 图1图的实验设计，使用空调向前配对。见表1本图中所使用的术语的定义。（一）政务司司长（气味）和之前与美国（蔗糖/水溶液）重叠。所示的CS和美国之间的关系是最适合调理蜜蜂和飞蛾。但最佳的ISI可以取决于空调协议和动物物种1（B）击穿和CS，CR，美国，和UR的相对定时。这既发生前美国演讲或在非增强型“测试”试验的CR是实验方案的相关措施。 <img alt="图2" fo:content-width="5in" src="/files/ftp_upload/51057/51057fig2highres.jpg" width="500" /> 图2：气味枯竭与墨盒再利用。对于这些数据，气味盒是建立与己醇，如文中所述。己醇是常用的一种CS的PER研究;但是，消耗的实际速率将取决于气味和溶剂。墨盒，然后使用一次，持续​​4秒，每分钟超过800个试验（假设5蜜蜂/审判标准的实验，就等于40的用途）。在指定的溶剂中稀释，这是为每实验相对标准，分别为2.0M的（A）和0.2M的（B）（A，B）中的气味后，第一和第八试验通过吸附于固相上进行取样微萃取（SPME）纤维是当时脱附到气相色谱仪。根据峰的相对面积的第一次试验相对于第八两种溶剂后和两个浓度较高（秩配对符号秩检验p &lt;0.05 [*]或更大），这表明较少的气味被从暗盒传送后盘盒已被使用的次数（C，D）示出刚刚在第八TR相同的数据<html> IAL与y轴展开。它表明，耗尽是稍大于相对于矿物油的己烷，但仅在较低的浓度。 <img alt="图3" fo:content-width="5in" src="/files/ftp_upload/51057/51057fig3highres.jpg" width="500" /> 图3采集和测试用癸醛作为CS和方法2调理。 （一）试行按试验在12前锋配对试验（ 图1）缓解（CR）。数据来自空调四组，每组5蜜蜂N = 20蜜蜂。 ISI的是3秒和ITI为6分（B）的使用，从癸不同碳链长度和/或羰基的碳的位置的气味非增强型测试。气味被提出在横跨四个组的随机序列，并且为了避免在CR的消光分别穿插着增强试验用癸醛（每两至三次试验）。 e_content“FO：保together.within页=”总是“&gt; <img alt="图4" fo:content-width="5in" src="/files/ftp_upload/51057/51057fig4highres.jpg" width="500" /> 从车间图4的数据在此期间，研究生进行了培训 ，从三组条件，以辛醇蜜蜂用每个方法一，（一）数据采集（N = 23和15的全球定位系统1和2，分别） 来调节蜜蜂和甲基环己酮组（n = 10），（二）在未增强的测试，两者气味的CR。四项试验中随机序列各组提出。（三）非增强型测试，既气味两组蜜蜂，一个正向的配对（1的Gp，N = 23）和一个落后的配对组（n = 25）。 <table border="0" cellpadding="0" cellspacing="0"><tbody><tr><td> 期限（缩写） </td><td> 定义 </td><td> 相关的例子 </td></tr><tr><td>条件刺激（CS） </td><td> á刺激是引发很少或没有回应，在第一和美国发出信号</td><td>气味</td></tr><tr><td>条件反应（CR）一个</td><td>与美国关联之后到CS的响应</td><td>长鼻延长</td></tr><tr><td>非条件刺激（美国） </td><td>这引起了响应。一种生物学显著刺激</td><td>蔗糖/水溶液</td></tr><tr><td>无条件响应（UR）一个</td><td>与美国的反应</td><td>长鼻延长</td></tr><tr><td>各国刺激间隔（ISI） </td><td>发病的CS的美国和发作之间的时间</td><td>气味蔗糖区间</td></tr><tr><td>跨试用期（ITI） </td><td>单个动物相继CS美配对之间的时间</td><td>气味（试行n）的气味（试行N + 1）的时间间隔</td></tr></tbody></table> 表1林portant术语。请参考图1对这些术语的说明a。对于每调理CR和UR是相同的，虽然其他类型的关联和反射对CR和UR可以不同。 <a href="/files/ftp_upload/51057/BEE_4.avi" target="_blank">电影1。</a>蜂蜜蜜蜂每执行后，它已习惯于以一种气味作出反应的视频。注意蜜蜂的1.5秒​​以下的气味发作大约每响应。跟踪软件映射触角的运动响应呈现的气味。绘制在视频的右上角的线条描绘的实际触角运动。两个图形在上X（左）和Y的屏幕显示触角运动的底部（右）坐标。图中的垂直线，在视频显示表明，从左到右，光开，PER，不亮和长鼻回缩。

Watch this Scientific Journal Video about A Proboscis Extension Response Protocol for Investigating Behavioral Plasticity in Insects: Application to Basic, Biomedical, and Agricultural Research at JoV

A Proboscis Extension Response Protocol for Investigating Behavioral Plasticity in Insects: Application to Basic, Biomedical, and Agricultural Research

The Proboscis Extension Response (PER) conditioning protocol, developed for the honey bee (Apis mellifera), provides an ecologically-relevant and easily quantifiable means for studying several different mechanisms of learning in many insect species.

长鼻扩展响应协议为侦查行为可塑性的昆虫：应用基础，生物医学和农业研究

Insects modify their responses to stimuli through experience of associating those stimuli with events important for survival (e.g., food, mates, threats). ...

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Research

JoVE Journal

Neuroscience

33.5K 次观看.  Arizona State University. The Proboscis Extension Response (PER) conditioning protocol, developed for the honey bee (Apis mellifera), provides an ecologically-relevant and easily quantifiable means for studying several different mechanisms of learning in many insect species. 

视频: A Proboscis Extension Response Protocol for Investigating Behavioral Plasticity in Insects: Application to Basic, Biomedical, and Agricultural Research

Combining Behavioral Endocrinology and Experimental Economics: Testosterone and Social Decision Making

Behavioral endocrinological research in humans as well as in animals suggests that testosterone plays a key role in social interactions. Studies in rodents have shown a direct link between testosterone and aggressive behavior1 and folk wisdom adapts these findings to humans, suggesting that testosterone induces antisocial, egoistic or even aggressive behavior2. However, many researchers doubt a direct testosterone-aggression link in humans, arguing instead that testosterone is primarily involved in status-related behavior3,4. As a high status can also be achieved by aggressive and antisocial means it can be difficult to distinguish between anti-social and status seeking behavior.
We therefore set up an experimental environment, in which status can only be achieved by prosocial means. In a double-blind and placebo-controlled experiment, we administered a single sublingual dose of 0.5 mg of testosterone (with a hydroxypropyl-&beta;-cyclodextrin carrier) to 121 women and investigated their social interaction behavior in an economic bargaining paradigm. Real monetary incentives are at stake in this paradigm; every player A receives a certain amount of money and has to make an offer to another player B on how to share the money. If B accepts, she gets what was offered and player A keeps the rest. If B refuses the offer, nobody gets anything. A status seeking player A is expected to avoid being rejected by behaving in a prosocial way, i.e. by making higher offers.
The results show that if expectations about the hormone are controlled for, testosterone administration leads to a significant increase in fair bargaining offers compared to placebo. The role of expectations is reflected in the fact that subjects who report that they believe to have received testosterone make lower offers than those who say they believe that they were treated with a placebo. These findings suggest that the experimental economics approach is sensitive for detecting neurobiological effects as subtle as those achieved by administration of hormones. Moreover, the findings point towards the importance of both psychosocial as well as neuroendocrine factors in determining the influence of testosterone on human social behavior.

The procedure described in this protocol shows that testosterone administration and folk beliefs about testosterone may be associated with directly opposed social behaviors.

睾酮和社会决策，结合行为内分泌和实验经济学：

Behavioral endocrinological research in humans as well as in animals suggests that testosterone plays a key role in social interactions. Studies in ...

科研

神经科学

TMS: Using the Theta-Burst Protocol to Explore Mechanism of Plasticity in Individuals with Fragile X Syndrome and Autism

Fragile X Syndrome (FXS), also known as Martin-Bell Syndrome, is a genetic abnormality found on the X chromosome.1,2 Individuals suffering from FXS display abnormalities in the expression of FMR1 - a protein required for typical, healthy neural development.3 Recent data has suggested that the loss of this protein can cause the cortex to be hyperexcitable thereby affecting overall patterns of neural plasticity.4,5
In addition, Fragile X shows a strong comorbidity with autism: in fact, 30% of children with FXS are diagnosed with autism, and 2 - 5% of autistic children suffer from FXS.6
Transcranial Magnetic Stimulation (a non-invasive neurostimulatory and neuromodulatory technique that can transiently or lastingly modulate cortical excitability via the application of localized magnetic field pulses 7,8) represents a unique method of exploring plasticity and the manifestations of FXS within affected individuals. More specifically, Theta-Burst Stimulation (TBS), a specific stimulatory protocol shown to modulate cortical plasticity for a duration up to 30 minutes after stimulation cessation in healthy populations, has already proven an efficacious tool in the exploration of abnormal plasticity.9,10 
Recent studies have shown the effects of TBS last considerably longer in individuals on the autistic spectrum - up to 90 minutes.11 This extended effect-duration suggests an underlying abnormality in the brain's natural plasticity state in autistic individuals - similar to the hyperexcitability induced by Fragile X Syndrome.
In this experiment, utilizing single-pulse motor-evoked potentials (MEPs) as our benchmark, we will explore the effects of both intermittent and continuous TBS on cortical plasticity in individuals suffering from FXS and individuals on the Autistic Spectrum.

In this article, we examine the effects of Theta-Burst TMS stimulation on cortical plasticity in individuals suffering from Fragile X syndrome and individuals on the autistic spectrum.

TMS：使用的Theta突发协议，以探索个人与脆性X综合征与孤独症的可塑性Mechasnism

Fragile X Syndrome (FXS), also known as Martin-Bell Syndrome, is a genetic abnormality found on the X chromosome.1,2 Individuals suffering from FXS ...

Behavioural Pharmacology in Classical Conditioning of the Proboscis Extension Response in Honeybees (Apis mellifera)

Honeybees (Apis mellifera) are well known for their communication and orientation skills and for their impressive learning capability1,2. Because the survival of a honeybee colony depends on the exploitation of food sources, forager bees learn and memorize variable flower sites as well as their profitability. Forager bees can be easily trained in natural settings where they forage at a feeding site and learn the related signals such as odor or color. Appetitive associative learning can also be studied under controlled conditions in the laboratory by conditioning the proboscis extension response (PER) of individually harnessed honeybees3,4. This learning paradigm enables the study of the neuronal and molecular mechanisms that underlie learning and memory formation in a simple and highly reliable way5-12. A behavioral pharmacology approach is used to study molecular mechanisms. Drugs are injected systemically to interfere with the function of specific molecules during or after learning and memory formation13-16.
Here we demonstrate how to train harnessed honeybees in PER conditioning and how to apply drugs systemically by injection into the bee flight muscle.

Behavioural Pharmacology in Classical Conditioning of the Proboscis Extension Response in Honeybees Apis mellifera

We demonstrate how to implement a behavioral pharmacology method in an appetitive olfactory conditioning paradigm in honeybees (Apis mellifera) by systemic application of drugs. This method allows investigation of the mechanisms underlying learning and memory formation in a simple and reliable way.

在蜜蜂的长鼻扩展响应的经典条件反射行为药理学（蜜蜂）

Honeybees (Apis mellifera) are well known for their communication and orientation skills and for their impressive learning capability1,2. Because the ...

Enabling High Grayscale Resolution Displays and Accurate Response Time Measurements on Conventional Computers

Display systems based on conventional computer graphics cards are capable of generating images with 8-bit gray level resolution. However, most experiments in vision research require displays with more than 12 bits of luminance resolution. Several solutions are available. Bit++ 1 and DataPixx 2 use the Digital Visual Interface (DVI) output from graphics cards and high resolution (14 or 16-bit) digital-to-analog converters to drive analog display devices. The VideoSwitcher 3 described here combines analog video signals from the red and blue channels of graphics cards with different weights using a passive resister network 4 and an active circuit to deliver identical video signals to the three channels of color monitors. The method provides an inexpensive way to enable high-resolution monochromatic displays using conventional graphics cards and analog monitors. It can also provide trigger signals that can be used to mark stimulus onsets, making it easy to synchronize visual displays with physiological recordings or response time measurements.
Although computer keyboards and mice are frequently used in measuring response times (RT), the accuracy of these measurements is quite low. The RTbox is a specialized hardware and software solution for accurate RT measurements. Connected to the host computer through a USB connection, the driver of the RTbox is compatible with all conventional operating systems. It uses a microprocessor and high-resolution clock to record the identities and timing of button events, which are buffered until the host computer retrieves them. The recorded button events are not affected by potential timing uncertainties or biases associated with data transmission and processing in the host computer. The asynchronous storage greatly simplifies the design of user programs. Several methods are available to synchronize the clocks of the RTbox and the host computer. The RTbox can also receive external triggers and be used to measure RT with respect to external events. 
Both VideoSwitcher and RTbox are available for users to purchase. The relevant information and many demonstration programs can be found at <a href="http://lobes.usc.edu/">http://lobes.usc.edu/</a>.

Conventional computer hardware can not generate visual stimuli with sufficiently high grayscale resolution and measure response times with sufficient accuracy. We describe how to use the VideoSwitcher to produce high-resolution monochromatic displays, and the RTbox to measure response times with high accuracy on conventional computer hardware.

启用高灰度分辨率的显示器和传统计算机准确的响应时间测量

Display systems based on conventional computer graphics cards are capable of generating images with 8-bit gray level resolution. However, most experiments ...

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 ...

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 ...

Inducing Plasticity of Astrocytic Receptors by Manipulation of Neuronal Firing Rates

Close to two decades of research has established that astrocytes in situ and in vivo express numerous G protein-coupled receptors (GPCRs) that can be stimulated by neuronally-released transmitter. However, the ability of astrocytic receptors to exhibit plasticity in response to changes in neuronal activity has received little attention. Here we describe a model system that can be used to globally scale up or down astrocytic group I metabotropic glutamate receptors (mGluRs) in acute brain slices. Included are methods on how to prepare parasagittal hippocampal slices, construct chambers suitable for long-term slice incubation, bidirectionally manipulate neuronal action potential frequency, load astrocytes and astrocyte processes with fluorescent Ca2+ indicator, and measure changes in astrocytic Gq GPCR activity by recording spontaneous and evoked astrocyte Ca2+ events using confocal microscopy. In essence, a &#8220;calcium roadmap&#8221; is provided for how to measure plasticity of astrocytic Gq GPCRs. Applications of the technique for study of astrocytes are discussed. Having an understanding of how astrocytic receptor signaling is affected by changes in neuronal activity has important implications for both normal synaptic function as well as processes underlying neurological disorders and neurodegenerative disease.

Here we describe an adaptation of protocols used to induce homeostatic plasticity in neurons for the study of plasticity of astrocytic G protein-coupled receptors. Recently used to examine changes in astrocytic group I mGluRs in juvenile mice, the method can be applied to measure scaling of various astrocytic GPCRs, in tissue from adult mice in situ and in vivo, and to gain a better appreciation of the sensitivity of astrocytic receptors to changes in neuronal activity.

诱导星形胶质细胞受体的可塑性由神经元的放电频率操作

Close to two decades of research has established that astrocytes in situ and in vivo express numerous G protein-coupled receptors (GPCRs) that can be ...

Dyeing Insects for Behavioral Assays: the Mating Behavior of Anesthetized Drosophila

Mating experiments using Drosophila have contributed greatly to the understanding of sexual selection and behavior. Experiments often require simple, easy and cheap methods to distinguish between individuals in a trial. A standard technique for this is CO2 anaesthesia and then labelling or wing clipping each fly. However, this is invasive and has been shown to affect behavior. Other techniques have used coloration to identify flies. This article presents a simple and non-invasive method for labelling Drosophila that allows them to be individually identified within experiments, using food coloring. This method is used in trials where two males compete to mate with a female. Dyeing allowed quick and easy identification. There was, however, some difference in the strength of the coloration across the three species tested. Data is presented showing the dye has a lower impact on mating behavior than CO2 in Drosophila melanogaster. The impact of CO2 anaesthesia is shown to depend on the species of Drosophila, with D. pseudoobscura and D. subobscura showing no impact, whereas D. melanogaster males had reduced mating success. The dye method presented is applicable to a wide range of experimental designs.

Dyeing Insects for Behavioral Assays: the Mating Behavior of Anesthetized Drosophila

This protocol describes a simple, cost effective way to individually identify Drosophila or other insects. Demonstration data investigating mating success across three species of Drosophila show that this method is comparable or better than the use of CO2 anaesthesia.

染色昆虫行为检测：麻醉的交配行为<em&gt;果蝇</em

Mating experiments using Drosophila have contributed greatly to the understanding of sexual selection and behavior. Experiments often require simple, easy ...

Online Transcranial Magnetic Stimulation Protocol for Measuring Cortical Physiology Associated with Response Inhibition

We describe the development of a reproducible, child-friendly motor response inhibition task suitable for online Transcranial Magnetic Stimulation (TMS) characterization of primary motor cortex (M1) excitability and inhibition. Motor response inhibition prevents unwanted actions and is abnormal in several neuropsychiatric conditions. TMS is a non-invasive technology that can quantify M1 excitability and inhibition using single- and paired-pulse protocols and can be precisely timed to study cortical physiology with high temporal resolution. We modified the original Slater-Hammel (S-H) stop signal task to create a &#34;racecar&#34; version with TMS pulses time-locked to intra-trial events. This task is self-paced, with each trial initiating after a button push to move the racecar towards the 800 ms target. GO trials require a finger-lift to stop the racecar just before this target. Interspersed randomly are STOP trials (25%) during which the dynamically adjusted stop signal prompts subjects to prevent finger-lift. For GO trials, TMS pulses were delivered at 650 ms after trial onset; whereas, for STOP trials, the TMS pulses occurred 150 ms after the stop signal. The timings of the TMS pulses were decided based on electroencephalography (EEG) studies showing event-related changes in these time ranges during stop signal tasks. This task was studied in 3 blocks at two study sites (n=38) and we recorded behavioral performance and event-related motor-evoked potentials (MEP). Regression modelling was used to analyze MEP amplitudes using age as a covariate with multiple independent variables (sex, study site, block, TMS pulse condition [single- vs. paired-pulse], trial condition [GO, successful STOP, failed STOP]). The analysis showed that TMS pulse condition (p&#60;0.0001) and its interaction with trial condition (p=0.009) were significant. Future applications for this online S-H/TMS paradigm include the addition of simultaneous EEG acquisition to measure TMS-evoked EEG potentials. A potential limitation is that in children, the TMS pulse sound could&#160;affect behavioral task performance.

We describe an experimental procedure to quantify excitability and inhibition of primary motor cortex during a motor response inhibition task by using Transcranial Magnetic Stimulation throughout the course of a Stop Signal Task.

在线经颅磁刺激测量与反应抑制相关的皮质生理

We describe the development of a reproducible, child-friendly motor response inhibition task suitable for online Transcranial Magnetic Stimulation (TMS) ...

SwarmSight: Real-time Tracking of Insect Antenna Movements and Proboscis Extension Reflex Using a Common Preparation and Conventional Hardware

Many scientifically and agriculturally important insects use antennae to detect the presence of volatile chemical compounds and extend their proboscis during feeding. The ability to rapidly obtain high-resolution measurements of natural antenna and proboscis movements and assess how they change in response to chemical, developmental, and genetic manipulations can aid the understanding of insect behavior. By extending our previous work on assessing aggregate insect swarm or animal group movements from natural and laboratory videos using the video analysis software SwarmSight, we developed a novel, free, and open-source software module, SwarmSight Appendage Tracking (SwarmSight.org) for frame-by-frame tracking of insect antenna and proboscis positions from conventional web camera videos using conventional computers. The software processes frames about 120 times faster than humans, performs at better than human accuracy, and, using 30 frames per second (fps) videos, can capture antennal dynamics up to 15 Hz. The software was used to track the antennal response of honey bees to two odors and found significant mean antennal retractions away from the odor source about 1 s after odor presentation. We observed antenna position density heat map cluster formation and cluster and mean angle dependence on odor concentration.

This protocol describes steps for using the novel software, SwarmSight, for frame-by-frame tracking of insect antenna and proboscis positions from conventional web camera videos using conventional computers. The free, open-source software processes frames about 120 times faster than humans and performs at better than human accuracy.

SwarmSight: 利用共同的准备和常规硬件实时跟踪昆虫的天线运动和喙的延伸反射

Many scientifically and agriculturally important insects use antennae to detect the presence of volatile chemical compounds and extend their proboscis ...