这项工作是由一个NRSA博士前补助1F31DC012985（到RD）和美国国立卫生研究院资助，以支持JC 我们要感谢琳内亚魏斯博士的帮助编制数字上的稿子，约瑟夫·莱恩博士有益的意见和弗雷德里克博士马里昂轮询的有益的技术咨询。我们也想感谢的四审稿意见非常有用。

以下协议符合美国耶鲁大学的所有动物护理指引。 1。试剂和设备的准备<ol><li>录音设备设置（ 图1A）。 <ol><li fo:keep-together.within-page="always">选择是无大的变化温度或湿度，并从分离的电气和机械噪声，如冰箱和离心机的来源一个房间钻机安装。 <img alt="图1" fo:content-width="5in" fo:src="/files/ftp_upload/51355/51355fig1highres.jpg" src="/files/ftp_upload/51355/51355fig1.jpg" /> 图1。记录台设置的（一）概述。立体显微镜（一个）安装在隔振台（b）中选择参考电极支架（三）被安装在平台上的探头（四）相反，通过显微操作。一个出口的塑料管（五）提供针对飞准备加湿空气流也被安装在吨他的平台。该探头被连接到所述放大器（f）中，其连接到数字采集系统（DAS）（克），它被连接到PC（小时）。电极和出口管（B）配置:左侧参比电极，在右边记录电极，和空气流出口管冲着飞准备<a href="https://www.jove.com/files/ftp_upload/51355/51355fig1highres.jpg" target="_blank">点击这里查看大图。</a> </li><li>山体视显微镜抗振动台或平台的中心。 </li><li>附加显微操作为参比电极/昆虫制剂和探头/记录电极的显微镜的左侧和右侧，分别使用磁性台灯。 </li><li>贴装插座塑料管中的第三微操纵器到显微镜的后部，定向成使得管口指向朝向蝇预备的位置（参见图1B）。 </li><li>使用柔性塑料管，在TACH出口塑料管保温瓶部分装满了水。连接一个小水族馆泵到空气泡通过水的烧瓶中，通过对苍蝇的出口塑料管生成加湿空气流。 </li><li>摩光纤光源关闭振动台，定向输出经由正下方的编制一张白色卡片纸反射光照亮的准备。确保光源不休息在桌子上。注意:反映在纸张盘上的光源的优点是双重的:它能够使对比度提高，使得感受器更容易地可视化，并且它防止了准备，会导致直接的光加热。 </li><li>塞tastePROBE放大器进入数字采集系统（DAS）和DAS到个人计算机，根据供应商的使用说明书。插上脚踏开关触发，并在工作区下的安排。注:电气隔离墙壁插座放大器和DAS是高度DESIR能。 </li><li>用鳄鱼夹和绝缘电线和电工胶带长度连接的金属部件，以表电气接地显微镜，显微操作，以及光源。通过连接到地面建筑物或DAS，它是通过电源插头接地电接地的金属平台。 </li><li>供选择的个人电脑上的DAS安装相应的采集软件。注意:确保数字采集驱动程序都与PC上的操作系统兼容。 </li><li>配置软件的扩增（10到100倍），信号滤波（典型的是贝塞尔带通滤波器设定为100赫兹-3，000赫兹）和采样率（至少10千赫）。注:从味觉神经元的信号幅度一般在0.5-2毫伏范围内，所以显示比例设置，以方便他们的可视化。注意:100赫兹滤波器有助于排除外来的电噪声，但是，它的变化尖峰的形状，可以使先进的尖峰排序莫尔Ë挑战。可替换地，一个1赫兹滤波器都可以使用。 </li><li>任选地，法拉第笼可以被设置围绕整个振动台。然而，铝箔的小片通常是足够的，以减少由外部环境或调查所产生的任何噪声。 </li></ol></li><li fo:keep-together.within-page="always">玻璃电极的制备 <img alt="图2" fo:content-width="5in" fo:src="/files/ftp_upload/51355/51355fig2highres.jpg" src="/files/ftp_upload/51355/51355fig2.jpg" /> 图2。参考和记录电极。下玻璃毛细管拉成参比电极，用（A）的放大倍率和无（B）尖折断的照片，并记录电极（C）。白条表示2毫米。 <a href="https://www.jove.com/files/ftp_upload/51355/51355fig2highres.jpg" target="_blank">点击这里查看大图。</a> <ol><li>拉参考电极从使用吸管牵拉仪器的玻璃毛细管。注:移液器拉马计划的确切设置会有所不同仪器之间。努力实现一个很长的渐进的锥度。孔径在前端不是关键的，因为尖将飞准备（ 图2A和2B）之前被破坏，但是，确保该电极的锥形长度的直径既不太薄，这将不允许足够固定化的唇瓣，也不太大，这可能会损坏味觉神经元或破裂的唾液腺。 </li><li>从与灯丝用吸管拉马仪器硼硅玻璃毛细管拉记录电极。试图实现一个锥度比的参比电极的浅，约10-15微米（ 图2C）28的孔径。 </li></ol></li><li>促味剂的溶液的制备<ol><li>使用比德尔 - 弗鲁西城林格氏溶液化（B＆E）为参比电极的电解质。一公升的B＆E中，溶解7.5克氯化钠加入0.35g KCl和0.279克氯化钙2∙2H 2 O一公升超纯水。存储小等分试样在-20℃下</li><li>用30mM的tricholine柠檬酸溶液（TCC）作为记录电极电解质和溶剂为促味剂溶液25，如苦味或糖GRN响应是被测定。或者，1-3 73mM氯化钾溶液可以使用，如果是水电池的反应来测定。 </li><li>为了使促味剂的解决方案，权衡促味剂适量粉末状，加入台泥作出初始库存的浓度。使用该项进行系列稀释从这个初始库存，以产生用于测试所需的浓度。注意:如果促味剂不容易溶于水，其它溶剂，如乙醇，可用于使初始原料浓度。台泥适当控制解决方案，并没有促味剂的溶剂应该在这种情况下被使用。 </li><li>店等分长期在-20°C存储的促味剂溶液1工作等分试样在4℃下，用于记录使用长达一周，这取决于促味剂的化学性质。 </li></ol></li></ol> 2， 果蝇准备<img alt="图3" fo:content-width="5in" fo:src="/files/ftp_upload/51355/51355fig3highres.jpg" src="/files/ftp_upload/51355/51355fig3.jpg" /> 图3。录音准备工作苍蝇（A）插入参比电极的位置成飞背胸部。白色箭头指示的参比电极。 （ 二）参比电极的中间位置是:通过颈部和头部先进，长鼻尚未扩展。 （C，D）与参比电极粉煤灰在最后的位置与尖端内唇瓣电极，和长鼻完全展开。 <a href="https://www.jove.com/files/ftp_upload/51355/51355fig3highres.jpg" target="_blank">点击这里查看大图。</a> <ol><li>收集新eclosed苍蝇从维护良好的动态文化，温度和湿度控制的条件下生长记录，并把它们的年龄5-10天的记录之前，新鲜的培养瓶。 </li><li>准备飞行前寒意镜板置于冰上15-30分钟。 </li><li>与B＆E解决方案使用的是长而细胶针0.5毫米直径，如脊髓针1毫升注射器回填玻璃参比电极，并轻轻拍打任何气泡。打破少量尖关闭使用镊子和利用毛细作用来绘制出所有剩余的气泡组织，解剖显微镜下观察。 </li><li>滑块B＆E填充参比电极丝上参比电极座，注意不要引入气泡。 </li><li>吸飞入P200的移液器吸头，使用内置吸引苍蝇从管材，网格，和移液器吸头，在冰桶和冷却30-60秒29的地方。 </li><li>从冰取出显微镜片，擦去水分，并在显微镜下的位置。轻轻拍打飞出枪头到显微镜板。 注:飞应充分固定，以轻松操控。 </li><li>在低倍率，轻轻地用一对镊子取出前腿，同时保持稳定的胸部与其他钳子。放置在其腹侧飞，背部朝上。注意:请务必小心，以免在准备过程中接触唇瓣与钳在任何时候，以尽量减少机械损伤。 </li><li>按住苍蝇的地方，一对镊子，将参比电极的后背胸部正中线。条目的一个建议的角度大约为45度，在所述头部（ 图3A）的方向。 </li><li>固定参考electrodË持有人与橡皮泥，使得飞是在显微镜高倍下可见。通过颈部和头部操纵和角度的玻璃电极，通过使用两对钳子的滑动对参考电极架飞。注:很快，但工作顺利，它是更容易完成这个步骤，而苍蝇仍是从冷（ 图3B）固定。 </li><li>轻轻地延长长鼻用一对钳子的，而滑动蝇进一步向下玻璃参比电极，直到电极的前端是唇瓣内侧和长鼻完全伸展（ 图3C和3D）。注:小心不要刺破任何部分长鼻组织或扩张与参比电极的唇瓣的边缘，因为这样可能会损坏苍蝇和/或味觉神经元，影响录音质量。 </li></ol> 3。从Labellar感器记录 图4。从苍蝇录音（A）唇瓣上飞准备离开与接触的权利，在高倍率下对齐记录电极。 （ 二）记录电极和唇瓣单感器接触，在高放大倍率。 <a href="https://www.jove.com/files/ftp_upload/51355/51355fig4highres.jpg" target="_blank">点击这里查看大图。</a> <ol><li>总是通过触摸抗振动台或平台的金属表面录制过程中接触任何设备之前，自己接地！注意:不提供静电荷的探头为可损坏电路是极其重要的。 </li><li>安全参考电极夹到微操纵安装在recordin的航空表克钻机。位置1瓣唇瓣在视场显微镜的，在高倍率下（通常至少140X），并在加湿的空气的流线。 </li><li>开启加湿空气流，电脑，DAS和放大器。开放的采集软件。 </li><li>冲洗和填充玻璃记录电极与所需的促味剂。 <ol><li>通过使用注射器和塑料管28穿过管至少10次拉少量水冲洗玻璃记录电极用超纯水。 </li><li>冲洗记录电极与促味剂的至少5倍。填写记录电极大约有三分之一到一半充满的促味剂和管取出。如果有气泡，点击释放或干脆重新填充电极。 </li><li>滑动电极到探头的银线迅速而顺利以便不引入气泡。 </li></ol></li><li>刺激单感器与促味剂的填充记录电极。 <ol><li>使用micromanipu振荡器带给感兴趣感器对准记录电极。 </li><li>按下脚踏开关触发放大器的采集模式。 </li><li>超前的记录​​电极与显微操作的精细控制旋钮仔细直至它与感器和记录开始针尖接触。 </li><li>经过1-2秒取出电极。 </li><li>如果需要，重复步骤3.5与其他感器。注:等待至少1分钟的演讲到同一感器之间。如果有用于长时间的单个促味剂的记录，所述促味剂的溶液干燥后会在尖端的溶液可能会变得更加集中。这可通过用平滑的纸张通过毛细管作用来除去少量的液体玻璃电极的前端轻轻接触来补救。 </li></ol></li><li>记录对另一促味剂，冲洗并用新的促味剂负荷记录电极，然后重复步骤3.4。注:彻底冲洗味质的电极是ABSolutely至关重要的，以避免交叉污染。 </li><li>定期保存数据文件的标识信息，如日期，基因型和促味剂。注:重要的是要保持记录会话的数据分析过程中的促味剂和每个演示感器身份的书面记录是很重要的。 </li></ol>

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I., Davis, A., Ramaswamy, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Contribution+of+different+taste+cells+and+signaling+pathways+to+the+discrimination+of+"bitter"+taste+stimuli+by+an+insect.">Contribution of different taste cells and signaling pathways to the discrimination of "bitter" taste stimuli by an insect.</a> The Journal of neuroscience : the official journal of the Society for Neuroscience. 22, 7281-7287 (2002).</li><li>Sanford, J. L., Shields, V. D., Dickens, J. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Gustatory+receptor+neuron+responds+to+DEET+and+other+insect+repellents+in+the+yellow-fever+mosquito,+Aedes+aegypti.">Gustatory receptor neuron responds to DEET and other insect repellents in the yellow-fever mosquito, Aedes aegypti.</a> Die Naturwissenschaften. 100, 269-273 (2013).</li><li>Merivee, E., Must, A., Milius, M., Luik, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Electrophysiological+identification+of+the+sugar+cell+in+antennal+taste+sensilla+of+the+predatory+ground+beetle+Pterostichus+aethiops.">Electrophysiological identification of the sugar cell in antennal taste sensilla of the predatory ground beetle Pterostichus aethiops.</a> J Insect Physiol. 53, 377-384 (2007).</li><li>Popescu, A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Function+and+central+projections+of+gustatory+receptor+neurons+on+the+antenna+of+the+noctuid+moth+Spodoptera+littoralis.">Function and central projections of gustatory receptor neurons on the antenna of the noctuid moth Spodoptera littoralis.</a> Journal of comparative physiology. A, Neuroethology. 199, 403-416 (2013).</li><li>Marion-Poll, F., Der Pers, J. V. a. n. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Un-filtered+recordings+from+insect+taste+sensilla.">Un-filtered recordings from insect taste sensilla.</a> Entomologia Experimentalis et Applicata. 80, 113-115 (1996).</li><li>Wieczorek, H., Wolff, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+labellar+sugar+receptor+of+Drosophila.">The labellar sugar receptor of Drosophila.</a> J. Comp. Physiol. A. Neuroethol Sens. Neural Behav. Physiol. 164, 825-834 (1989).</li><li>Morita, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Initiation+of+spike+potentials+in+contact+chemosensory+hairs+of+insects.+III.+D.C.+stimulation+and+generator+potential+of+labellar+chemoreceptor+of+calliphora.">Initiation of spike potentials in contact chemosensory hairs of insects. III. D.C. stimulation and generator potential of labellar chemoreceptor of calliphora.</a> Journal of cellular and comparative physiology. 54, 189-204 (1959).</li><li>Lacaille, F., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=An+inhibitory+sex+pheromone+tastes+bitter+for+Drosophila+males.">An inhibitory sex pheromone tastes bitter for Drosophila males.</a> PLoS One. 2, e661 (2007).</li><li>Benton, R., Dahanukar, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Electrophysiological+recording+from+Drosophila+taste+sensilla.">Electrophysiological recording from Drosophila taste sensilla.</a> Cold Spring Harbor protocols. 2011, 839-850 (2011).</li><li>Pellegrino, M., Nakagawa, T., Vosshall, L. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Single+sensillum+recordings+in+the+insects+Drosophila+melanogaster+and+Anopheles+gambiae.">Single sensillum recordings in the insects Drosophila melanogaster and Anopheles gambiae.</a> J. Vis. Exp. , e1725 (2010).</li><li>Axon Instruments. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> The Axon Guide for Electrophysiology &amp; Biophysics Laboratory Techniques. , (1993).</li><li>Fujishiro, N., Kijima, H., Morita, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Impulse+frequency+and+action+potential+amplitude+in+labellar+chemosensory+neurones+of+Drosophila+melanogaster.">Impulse frequency and action potential amplitude in labellar chemosensory neurones of Drosophila melanogaster.</a> Journal of insect physiology. 30, 317-325 (1984).</li><li>Marion-Poll, F., Tobin, T. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Software+filter+for+detecting+spikes+superimposed+on+a+fluctuating+baseline.">Software filter for detecting spikes superimposed on a fluctuating baseline.</a> Journal of neuroscience. 37, 1-6 (1991).</li><li>Meunier, N., Marion-Poll, F., Lansky, P., Rospars, J. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Estimation+of+the+individual+firing+frequencies+of+two+neurons+recorded+with+a+single+electrode.">Estimation of the individual firing frequencies of two neurons recorded with a single electrode.</a> Chem Senses. 28, 671-679 (2003).</li><li>Meunier, N., Marion-Poll, F., Rospars, J. P., Tanimura, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Peripheral+coding+of+bitter+taste+in+Drosophila.">Peripheral coding of bitter taste in Drosophila.</a> Journal of neurobiology. 56, 139-152 (2003).</li></ol>

作者什么都没有透露。

Labellar感受器由于形态和解剖组织的差异而有所不同在便于记录。有时候感器不响应任何促味剂，连一个已知可引起积极的回应。取决于感器类型与发生这种情况的频率而变化。 L感器是最一致响应和相对容易访问，因为它们的长度。一般来说，S感器是一致响应，但他们的短长并在唇瓣位置好好接触挑战性。余感器可以更容易地访问，这取决于制剂的角度，但是，它们更频繁响应。在任何给定的?...

味觉允许的昆虫以检测可溶性的化学物质种类繁多并且起着一种营养物质的接受了重要的作用，或一种有毒或毒性1的抑制。味道也被认为在择偶中发挥作用，通过信息素1-5的检测。这些重要而多样化的功能所做的昆虫味觉系统调查的令人信服的目标进入感觉系统如何翻译环境因素纳入有关的行为输出。 果蝇味觉系统的主要单位是味道的头发，或感器。分子通过孔在其尖端2,6进入感器。感器上发现的唇瓣，腿，翅缘，咽6。在唇瓣，感受器的数量和位置是千篇一律。有感器基于长度3形态类:长（L），中间体（I），和短（S<html> ）感器7,8。每个感器包含两个（I-型）或4（L-和S-型）味觉受体神经元（GRNS）9。不同GRNS应对不同类别的味觉刺激的:苦，糖，盐和渗透压7,10和表达味觉受体8,11-13的不同子集。只有我和S型感器含有苦响应GRNS 8,10。该GRNS项目的subesophageal神经节（SOG）和其活化味分子被传递到用于解码的更高中枢神经系统，从而导致行为反应6。数量相对较少的神经元和顺从分子和行为分析使果蝇味觉系统的味觉系统的一般调查一个很好的模型。相对难易程度，系统可以通过基因突变或GAL4-UAS表达系统来操作也可作为一种有价值的工具14,15。 ontent"&gt;因为这些感受器从唇瓣的表面突出，它们使电生理学优良的目标。的GRNS的烧成可以使用外记录进行监控。历史上，侧壁的记录方法，其使用插入玻璃电极感器来记录神经细胞的活动，26已被使用，但是，这种方法在技术上是挑战性的执行，并且难以进行长时间录制从每个制备的针尖记录方法，其测量神经元的响应与电极的那同时提供了一个促味，从此成为首选9,16的方法，它已被用来研究果蝇 8,10,17,18的味觉系统，以及其他一些昆虫种类19-23，它有由tastePROBE放大器的发展，这克服了针尖记录方法的主要缺点之一通过补偿得到了极大的便利参比电极和对昆虫感器之间的大的电势差，使得没有过多的放大或过滤24要被记录的GRN动作电位。另一个重要发展是采用tricholine柠檬酸作为记录电解质25。台泥抑制反应的渗透压敏感GRN和不刺激盐敏感GRN，使得由苦和糖味质生成的反应更容易进行分析25。 在这里，我们描述了如何在目前卡尔森实验室进行的果蝇 labellar感受器尖记录。该协议将解释如何建立一个合适的电钻机，如何准备飞，以及如何执行味道录音。我们也当使用这种出示由果蝇感器的子集，以及一些常见的问题和可能遇到的潜在解决方案的记录得出了一些有代表性的数据技术。

<table border="0" cellpadding="0" cellspacing="0" width="783">
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	<tbody>
		<tr height="84">
			<td height="84" style="height:84px;width:168px;">Stereo Zoom Microscope</td>
			<td style="width:187px;">Olympus&#160;</td>
			<td style="width:211px;">SZX12 DFPLFL1.6x PF eyepieces: WHN10x-H/22</td>
			<td style="width:219px;">capable of ~150x magnification with long working distance table mount stand</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:168px;">Anti-vibration Table</td>
			<td style="width:187px;">Kinetic Systems</td>
			<td>BenchMate2210</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:168px;">Micromanipulators</td>
			<td style="width:187px;">Narishige</td>
			<td>NMN-21</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:168px;">Magnetic stands</td>
			<td style="width:187px;">ENCO</td>
			<td>Model #625-0930</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:168px;">Reference Electrode Holder</td>
			<td style="width:187px;">Harvard Apparatus</td>
			<td style="width:211px;">ESP/W-F10N</td>
			<td>Can be mounted on 5ml serological pipette for extended range</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:168px;">&#160;Silver Wire</td>
			<td style="width:187px;">World Precision Instruments</td>
			<td style="width:211px;">AGW1510</td>
			<td>0.3-0.5mm diameter</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:168px;">Retort Stand</td>
			<td style="width:187px;"></td>
			<td style="width:211px;"></td>
			<td>generic</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:168px;">Outlet Plastic Tube</td>
			<td style="width:187px;"></td>
			<td style="width:211px;"></td>
			<td>generic, 1cm diameter</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:168px;">Flexible Plastic Tubing</td>
			<td style="width:187px;">Nalgene&#160;</td>
			<td>8000-0060</td>
			<td style="width:219px;">VI grade 1/4 in internal diameter&#160;</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:168px;">500 ml Conical Flask</td>
			<td style="width:187px;"></td>
			<td style="width:211px;"></td>
			<td>generic,&#160; with side arm</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:168px;">Aquarium Pump</td>
			<td style="width:187px;">Aquatic Gardens</td>
			<td style="width:211px;">Airpump 2000</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:168px;">Fiber Optic Light Source</td>
			<td style="width:187px;">Dolan-Jenner Industries</td>
			<td>Fiber-Lite 2100</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:168px;">White Card/Paper</td>
			<td style="width:187px;">Whatman</td>
			<td style="width:211px;">1001-110</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:168px;">Digital Acquisition System</td>
			<td style="width:187px;">Syntech</td>
			<td style="width:211px;">IDAC-4</td>
			<td>Alternative: National Instruments NI-6251 &#160;</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:168px;">Headstage</td>
			<td style="width:187px;">Syntech</td>
			<td style="width:211px;">DTP-1</td>
			<td>Tasteprobe</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:168px;">Tasteprobe Amplifier</td>
			<td style="width:187px;">Syntech</td>
			<td style="width:211px;">DTP-1</td>
			<td>Tasteprobe</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:168px;">Alligator Clips</td>
			<td style="width:187px;">Grainger</td>
			<td style="width:211px;">1XWN7</td>
			<td>Any brand is fine</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:168px;">Insulated Electrical Wire</td>
			<td style="width:187px;"></td>
			<td style="width:211px;"></td>
			<td>Generic</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:168px;">Gold Connector Pins</td>
			<td style="width:187px;">World Precision Instruments</td>
			<td align="right" style="width:211px;">5482</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:168px;">Personal Computer</td>
			<td style="width:187px;">Dell&#160;</td>
			<td style="width:211px;">Vostro</td>
			<td>Check for compatibility with digital acquisition system and software</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:168px;">Acquisition Software</td>
			<td style="width:187px;">Syntech</td>
			<td style="width:211px;">Autospike</td>
			<td>Autospike works with IDAC-4; alternatively, use Labview with NI-6251</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:168px;">Aluminum Foil and/or Faraday Cage</td>
			<td style="width:187px;"></td>
			<td style="width:211px;"></td>
			<td>Electro-magnetic noise shielding</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:168px;"></td>
			<td style="width:187px;"></td>
			<td style="width:211px;"></td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:168px;">Borosilicate Glass Capillaries</td>
			<td style="width:187px;">World Precision Instruments</td>
			<td style="width:211px;">1B100F-4</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:168px;">Pipette Puller</td>
			<td style="width:187px;">Sutter Instrument Company</td>
			<td style="width:211px;">Model P-87 Flaming/Brown Micropipette Puller</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:168px;"></td>
			<td style="width:187px;"></td>
			<td style="width:211px;"></td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:168px;">Beadle and Ephrussi Ringer Solution</td>
			<td style="width:187px;"></td>
			<td style="width:211px;"></td>
			<td>See recipe in protocol section</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:168px;">Tricholine citrate, 65%&#160;</td>
			<td style="width:187px;">Sigma</td>
			<td style="width:211px;">T0252-100G</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:168px;"></td>
			<td style="width:187px;"></td>
			<td style="width:211px;"></td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:168px;">Stereo Microscope</td>
			<td style="width:187px;">Olympus</td>
			<td style="width:211px;">VMZ 1x-4x</td>
			<td style="width:219px;">Capable of 10x-40x magnification</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:168px;">Ice Bucket</td>
			<td style="width:187px;"></td>
			<td style="width:211px;"></td>
			<td>Generic</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:168px;">p200 Pipette Tips</td>
			<td style="width:187px;"></td>
			<td style="width:211px;"></td>
			<td>Generic</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:168px;">Spinal Needle</td>
			<td style="width:187px;">Terumo</td>
			<td style="width:211px;">SN*2590</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:168px;">1ml Syringe</td>
			<td style="width:187px;">Beckton-Dickenson</td>
			<td align="right" style="width:211px;">301025</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:168px;">Fly Aspirator</td>
			<td style="width:187px;"></td>
			<td style="width:211px;"></td>
			<td>Assembled from P1000 pipette tips, flexible plastic tubing, and mesh</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:168px;">Modeling Clay</td>
			<td style="width:187px;"></td>
			<td style="width:211px;"></td>
			<td>Generic</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:168px;">Forceps</td>
			<td style="width:187px;">Fine Science Tools By Dumont</td>
			<td style="width:211px;">11252-00</td>
			<td>#5SF (super-fine tips)</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:168px;">10ml Syringe&#160;</td>
			<td style="width:187px;">Beckton-Dickinson</td>
			<td align="right" style="width:211px;">301029</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:168px;">Plastic Tubing</td>
			<td style="width:187px;">Tygon</td>
			<td style="width:211px;">R-3603</td>
			<td></td>
		</tr>
	</tbody>
</table>

electrophysiological recording from drosophila labellar taste sensilla

昆虫的外围味道响应可以有力地调查与电生理技术。这里介绍的方法使研究人员直接和定量测量味觉反应，反映了感官输入，昆虫的神经系统从它的环境味觉刺激接收。该协议概述了在利用这项技术的所有关键步骤。在组装的电装备，如选择的必要设备和一个合适的环境记录的关键步骤，圈定。我们还描述了记录作出适当的参考和记录电极如何准备，并促味剂的解决方案。我们详细地描述用于通过插入玻璃参比电极成飞，以固定所述长鼻制备昆虫的方法。我们显示通过味觉神经元响应于一个糖和一个苦味化合物烧制的电脉冲的痕迹。该协议的某些方面为technically具有挑战性的，我们包括可能​​遇到的一些常见的技术挑战，如缺乏信号或噪音过大的系统，和潜在的解决方案广泛的描述。该技术具有局限性，如不能交付时间上复杂的刺激，观察背景发射前夕刺激交付，或使用不溶于水的呈味物质方便。尽管有这些限制，这项技术（包括在协议中引用的微小变化）是一种标准，用于记录果蝇神经元对味道的化合物广泛接受的过程。

图5A示出了一个L形感器的一种糖，蔗糖的反应。同一感器不给苦化合物反应，黄连， 图5B示出了一个I型感器，它包含一个苦响应神经元，显示响应于小檗碱较大的幅度峰值，并响应于蔗糖较小的幅度峰值。 L感器显示一个最小的背景响应溶剂对照，台泥，而我感器显示给TCC几乎没有响应（ 图5）。有关labellar GRNS的盐和水反应的更多信息，请参阅广井10。</...

Watch this Scientific Journal Video about Electrophysiological Recording From Drosophila Labellar Taste Sensilla at JoVE.com

Electrophysiological Recording From Drosophila Labellar Taste Sensilla

本协议描述的由labellar味觉神经元在果蝇发射动作电位反应细胞外记录。

电生理记录从<em&gt;果蝇</em&gt; Labellar味感器

The peripheral taste response of insects can be powerfully investigated with electrophysiological techniques. The method described here allows the ...

electrophysiological-recording-from-drosophila-labellar-taste-sensilla

Research

JoVE Journal

Neuroscience

Electrophysiological Recording From Drosophila Labellar Taste Sensilla

16.2K 次观看.  Yale University. 本协议描述的由labellar味觉神经元在果蝇发射动作电位反应细胞外记录。 

视频: Electrophysiological Recording From Drosophila Labellar Taste Sensilla

Simultaneous Recording of Calcium Signals from Identified Neurons and Feeding Behavior of Drosophila melanogaster

To study neuronal networks in terms of their function in behavior, we must analyze how neurons operate when each behavioral pattern is generated. Thus, simultaneous recordings of neuronal activity and behavior are essential to correlate brain activity to behavior. For such behavioral analyses, the fruit fly, Drosophila melanogaster, allows us to incorporate genetically encoded calcium indicators such as GCaMP1, to monitor neuronal activity, and to use sophisticated genetic manipulations for optogenetic or thermogenetic techniques to specifically activate identified neurons2-5. Use of a thermogenetic technique has led us to find critical neurons for feeding behavior (Flood et al., under revision). As a main part of feeding behavior, a Drosophila adult extends its proboscis for feeding6 (proboscis extension response; PER), responding to a sweet stimulus from sensory cells on its proboscis or tarsi. Combining the protocol for PER7 with a calcium imaging technique8 using GCaMP3.01, 9, I have established an experimental system, where we can monitor activity of neurons in the feeding center &#x2013; the suboesophageal ganglion (SOG), simultaneously with behavioral observation of the proboscis. I have designed an apparatus ("Fly brain Live Imaging and Electrophysiology Stage": "FLIES") to accommodate a Drosophila adult, allowing its proboscis to freely move while its brain is exposed to the bath for Ca2+ imaging through a water immersion lens. The FLIES is also appropriate for many types of live experiments on fly brains such as electrophysiological recording or time lapse imaging of synaptic morphology. Because the results from live imaging can be directly correlated with the simultaneous PER behavior, this methodology can provide an excellent experimental system to study information processing of neuronal networks, and how this cellular activity is coupled to plastic processes and memory.

Simultaneous Recording of Calcium Signals from Identified Neurons and Feeding Behavior of Drosophila melanogaster

The fruit fly, Drosophila melanogaster, extends its proboscis for feeding, responding to a sugar stimulus from its proboscis or tarsus. I have combined observations of the proboscis extension response (PER) with a calcium imaging technique, allowing us to monitor the activity of neurons in the brain, simultaneously with behavioral observation.

从查明的神经元和取食行为的钙信号的同时记录果蝇</em

To study neuronal networks in terms of their function in behavior, we must analyze how neurons operate when each behavioral pattern is generated.  Thus, ...

科研

神经科学

Forebrain Electrophysiological Recording in Larval Zebrafish

Epilepsy affects nearly 3 million people in the United States and up to 50 million people worldwide. Defined as the occurrence of spontaneous unprovoked seizures, epilepsy can be acquired as a result of an insult to the brain or a genetic mutation. Efforts to model seizures in animals have primarily utilized acquired insults (convulsant drugs, stimulation or brain injury) and genetic manipulations (antisense knockdown, homologous recombination or transgenesis) in rodents. Zebrafish are a vertebrate model system1-3 that could provide a valuable alternative to rodent-based epilepsy research. Zebrafish are used extensively in the study of vertebrate genetics or development, exhibit a high degree of genetic similarity to mammals and express homologs for ~85% of known human single-gene epilepsy mutations. Because of their small size (4-6 mm in length), zebrafish larvae can be maintained in fluid volumes as low as 100 &mu;l during early development and arrayed in multi-well plates. Reagents can be added directly to the solution in which embryos develop, simplifying drug administration and enabling rapid in vivo screening of test compounds4. Synthetic oligonucleotides (morpholinos), mutagenesis, zinc finger nuclease and transgenic approaches can be used to rapidly generate gene knockdown or mutation in zebrafish5-7. These properties afford zebrafish studies an unprecedented statistical power analysis advantage over rodents in the study of neurological disorders such as epilepsy. Because the "gold standard" for epilepsy research is to monitor and analyze the abnormal electrical discharges that originate in a central brain structure (i.e., seizures), a method to efficiently record brain activity in larval zebrafish is described here. This method is an adaptation of conventional extracellular recording techniques and allows for stable long-term monitoring of brain activity in intact zebrafish larvae. Sample recordings are shown for acute seizures induced by bath application of convulsant drugs and spontaneous seizures recorded in a genetically modified fish.

A simple method to record extracellular field potentials in the larval zebrafish forebrain is described. The method provides a robust in vivo read-out of seizure-like activity. This technique can be used with genetically modified zebrafish larvae carrying epilepsy-related genes or seizures evoked by administration of convulsant drugs.

斑马鱼幼虫的前脑的电生理记录

Epilepsy affects nearly 3 million people in the United States and up to 50 million people worldwide. Defined as the occurrence of spontaneous unprovoked ...

Simultaneous Pre- and Post-synaptic Electrophysiological Recording from Xenopus Nerve-muscle Co-cultures

Much information about the coupling of presynaptic ionic currents with the release of neurotransmitter has been obtained from invertebrate preparations, most notably the squid giant synapse1. However, except for the preparation described here, few vertebrate preparations exist in which it is possible to make simultaneous measurements of neurotransmitter release and presynaptic ionic currents. Embryonic Xenopus motoneurons and muscle cells can be grown together in simple culture medium at room temperature; they will form functional synapses within twelve to twenty-four hours, and can be used to study nerve and muscle cell development and synaptic interactions for several days (until overgrowth occurs). Some advantages of these co-cultures over other vertebrate preparations include the simplicity of preparation, the ability to maintain the cultures and work at room temperature, and the ready accessibility of the synapses formed2-4. The preparation has been used widely to study the biophysical properties of presynaptic ion channels and the regulation of transmitter release5-8. In addition, the preparation has lent itself to other uses including the study of neurite outgrowth and synaptogenesis9-12, molecular mechanisms of neurotransmitter release13-15, the role of diffusible messengers in neuromodulation16,17, and in vitro synaptic plasticity18-19.

Simultaneous Pre- and Post-synaptic Electrophysiological Recording from Xenopus Nerve-muscle Co-cultures

This video demonstrates the procedures used to grow primary cultures of embryonic Xenopus nerve and muscle cells and the usefulness of this preparation for making simultaneous pre- and post-synaptic patch clamp recordings.

同时前和突触后电生理记录<em&gt;爪蟾</em神经肌肉文化

Much information about the coupling of  presynaptic ionic currents with the release of neurotransmitter has been obtained  from invertebrate preparations, ...

Simultaneous Electrophysiological Recording and Calcium Imaging of Suprachiasmatic Nucleus Neurons

Simultaneous electrophysiological and fluorescent imaging recording methods were used to study the role of changes of membrane potential or current in regulating the intracellular calcium concentration. Changing environmental conditions, such as the light-dark cycle, can modify neuronal and neural network activity and the expression of a family of circadian clock genes within the suprachiasmatic nucleus (SCN), the location of the master circadian clock in the mammalian brain. Excitatory synaptic transmission leads to an increase in the postsynaptic Ca2+ concentration that is believed to activate the signaling pathways that shifts the rhythmic expression of circadian clock genes. Hypothalamic slices containing the SCN were patch clamped using microelectrodes filled with an internal solution containing the calcium indicator bis-fura-2. After a seal was formed between the microelectrode and the SCN neuronal membrane, the membrane was ruptured using gentle suction and the calcium probe diffused into the neuron filling both the soma and dendrites. Quantitative ratiometric measurements of the intracellular calcium concentration were recorded simultaneously with membrane potential or current. Using these methods it is possible to study the role of changes of the intracellular calcium concentration produced by synaptic activity and action potential firing of individual neurons. In this presentation we demonstrate the methods to simultaneously record electrophysiological activity along with intracellular calcium from individual SCN neurons maintained in brain slices.

Procedures are described to perform simultaneous recordings of membrane potential or current and changes of intracellular calcium concentration. Suprachiasmatic nucleus neurons are filled with the calcium indicator bis-fura-2 using a patch clamp electrode in the whole cell patch clamp configuration.

Simultaneous electrophysiological and fluorescent imaging recording methods were used to study the role of changes of membrane potential or current in ...

Membrane Potential Dye Imaging of Ventromedial Hypothalamus Neurons From Adult Mice to Study Glucose Sensing

Studies of neuronal activity are often performed using neurons from rodents less than 2 months of age due to the technical difficulties associated with increasing connective tissue and decreased neuronal viability that occur with age. Here, we describe a methodology for the dissociation of healthy hypothalamic neurons from adult-aged mice. The ability to study neurons from adult-aged mice allows the use of disease models that manifest at a later age and might be more developmentally accurate for certain studies. Fluorescence imaging of dissociated neurons can be used to study the activity of a population of neurons, as opposed to using electrophysiology to study a single neuron. This is particularly useful when studying a heterogeneous neuronal population in which the desired neuronal type is rare such as for hypothalamic glucose sensing neurons. We utilized membrane potential dye imaging of adult ventromedial hypothalamic neurons to study their responses to changes in extracellular glucose. Glucose sensing neurons are believed to play a role in central regulation of energy balance. The ability to study glucose sensing in adult rodents is particularly useful since the predominance of diseases related to dysfunctional energy balance (e.g.&#160;obesity) increase with age.

The activity of single neurons from adult-aged mice can be studied by dissociating neurons from specific brain regions and using fluorescent membrane potential dye imaging. By testing responses to changes in glucose, this technique can be used to study the glucose sensitivity of adult ventromedial hypothalamic neurons.

膜下丘脑腹内侧神经元的电位染料成像从成年小鼠为研究葡萄糖传感

Studies of neuronal activity are often performed using neurons from rodents less than 2 months of age due to the technical difficulties associated with ...

Electrophysiological Recording in the Brain of Intact Adult Zebrafish

Previously, electrophysiological studies in adult zebrafish have been limited to slice preparations or to eye cup preparations and electrorentinogram recordings. This paper describes how an adult zebrafish can be immobilized, intubated, and used for in vivo electrophysiological experiments, allowing recording of neural activity. Immobilization of the adult requires a mechanism to deliver dissolved oxygen to the gills in lieu of buccal and opercular movement. With our technique, animals are immobilized and perfused with habitat water to fulfill this requirement. A craniotomy is performed under tricaine methanesulfonate (MS-222; tricaine) anesthesia to provide access to the brain. The primary electrode is then positioned within the craniotomy window to record extracellular brain activity. Through the use of a multitube perfusion system, a variety of pharmacological compounds can be administered to the adult fish and any alterations in the neural activity can be observed. The methodology not only allows for observations to be made regarding changes in neurological activity, but it also allows for comparisons to be made between larval and adult zebrafish. This gives researchers the ability to identify the alterations in neurological activity due to the introduction of various compounds at different life stages.

This paper describes how an adult zebrafish can be immobilized, intubated, and used for in vivo electrophysiological experiments to allow recordings and manipulation of neural activity in an intact animal.

在完整的成年斑马鱼大脑电生理记录

Previously, electrophysiological studies in adult zebrafish have been limited to slice preparations or to eye cup preparations and electrorentinogram ...

Electrophysiological Method for Recording Intracellular Voltage Responses of Drosophila Photoreceptors and Interneurons to Light Stimuli In Vivo

Voltage responses of insect photoreceptors and visual interneurons can be accurately recorded with conventional sharp microelectrodes. The method described here enables the investigator to measure long-lasting (from minutes to hours) high-quality intracellular responses from single Drosophila R1-R6 photoreceptors and Large Monopolar Cells (LMCs) to light stimuli. Because the recording system has low noise, it can be used to study variability among individual cells in the fly eye, and how their outputs reflect the physical properties of the visual environment. We outline all key steps in performing this technique. The basic steps in constructing an appropriate electrophysiology set-up for recording, such as design and selection of the experimental equipment are described. We also explain how to prepare for recording by making appropriate (sharp) recording and (blunt) reference electrodes. Details are given on how to fix an intact fly in a bespoke fly-holder, prepare a small window in its eye and insert a recording electrode through this hole with minimal damage. We explain how to localize the center of a cell's receptive field, dark- or light-adapt the studied cell, and to record its voltage responses to dynamic light stimuli. Finally, we describe the criteria for stable normal recordings, show characteristic high-quality voltage responses of individual cells to different light stimuli, and briefly define how to quantify their signaling performance. Many aspects of the method are technically challenging and require practice and patience to master. But once learned and optimized for the investigator's experimental objectives, it grants outstanding in vivo neurophysiological data.

Electrophysiological Method for Recording Intracellular Voltage Responses of Drosophila Photoreceptors and Interneurons to Light Stimuli In Vivo

Sharp microelectrodes enable accurate electrophysiological characterization of photoreceptor and visual interneuron output in living Drosophila. Here we show how to use this method to record high-quality voltage responses of individual cells to controlled light stimulation. This method is ideal for studying neural information processing in insect compound eyes.

为记录细胞内电压响应电法<em&gt;果蝇</em&gt;光感受器和中间对光刺激<em&gt;在体内</em

Voltage responses of insect photoreceptors and visual interneurons can be accurately recorded with conventional sharp microelectrodes. The method ...

Taste Preference Assay for Adult Drosophila

Olfactory and gustatory perception of the environment is vital for animal survival. The most obvious application of these chemosenses is to be able to distinguish good food sources from potentially dangerous food sources. Gustation requires physical contact with a chemical compound which is able to signal through taste receptors that are expressed on the surface of neurons. In insects, these gustatory neurons can be located across the animal's body allowing taste to play an important role in many different behaviors. Insects typically prefer compounds containing sugars, while compounds that are considered bitter tasting are avoided. Given the basic biological importance of taste, there is intense interest in understanding the molecular mechanisms underlying this sensory modality. We describe an adult Drosophila taste assay which reflects the preference of the animals for a given tastant compound. This assay may be applied to animals of any genetic background to examine the taste preference for a desired soluble compound.

Taste Preference Assay for Adult Drosophila

Taste is an important sensory process which facilitates attraction to beneficial substances and avoidance of toxic substances. This protocol describes a simple ingestion assay for determining Drosophila gustatory preference for a given chemical compound.

口味偏好测定成人<em&gt;果蝇</em

Olfactory and gustatory perception of the environment is vital for animal survival.  The most obvious application of these chemosenses is to be able to ...

Electrophysiological Method for Whole-cell Voltage Clamp Recordings from Drosophila Photoreceptors

Whole-cell voltage clamp recordings from Drosophila melanogaster photoreceptors have revolutionized the field of invertebrate visual transduction, enabling the use of D. melanogaster molecular genetics to study inositol-lipid signaling and Transient Receptor Potential (TRP) channels at the single-molecule level. A handful of labs have mastered this powerful technique, which enables the analysis of the physiological responses to light under highly controlled conditions. This technique allows control over the intracellular and extracellular media; the membrane voltage; and the fast application of pharmacological compounds, such as a variety of ionic or pH indicators, to the intra- and extracellular media. With an exceptionally high signal-to-noise ratio, this method enables the measurement of dark spontaneous and light-induced unitary currents (i.e.&#160;spontaneous and quantum bumps) and macroscopic Light-induced Currents (LIC) from single D. melanogaster photoreceptors. This protocol outlines, in great detail, all the key steps necessary to perform this technique, which includes both electrophysiological and optical recordings. The fly retina dissection procedure for the attainment of intact and viable ex vivo isolated ommatidia in the bath chamber is described. The equipment needed to perform whole-cell and fluorescence imaging measurements are also detailed. Finally, the pitfalls in using this delicate preparation during extended experiments are explained.

Electrophysiological Method for Whole-cell Voltage Clamp Recordings from Drosophila Photoreceptors

Whole-cell recordings from Drosophila melanogaster photoreceptors enable the measurement of spontaneous dark bumps, quantum bumps, macroscopic responses to light, and current-voltage relationships under various conditions. In combination with D. melanogaster genetic manipulation tools, this method enables the study of the ubiquitous inositol-lipid signaling pathway and its target, the TRP channel.

全细胞电压钳记录的电生理方法<em&gt;果蝇</em&gt;光感受器

Whole-cell voltage clamp recordings from Drosophila melanogaster photoreceptors have revolutionized the field of invertebrate visual transduction, ...

Electrophysiological Recording from Drosophila Trichoid Sensilla in Response to Odorants of Low Volatility

Insects rely on their sense of smell to guide a wide range of behaviors that are critical for their survival, such as food-seeking, predator avoidance, oviposition, and mating. Myriad chemicals of varying volatilities have been identified as natural odorants that activate insect Olfactory Receptor Neurons (ORNs). However, studying the olfactory responses to low-volatility odorants has been hampered by an inability to effectively present such stimuli using conventional odor-delivery methods. Here, we describe a procedure that permits the effective presentation of low-volatility odorants for in vivo Single-Sensillum Recording (SSR). By minimizing the distance between the odor source and the target tissue, this method allows for the application of biologically salient but hitherto inaccessible odorants, including palmitoleic acid, a stimulatory pheromone with a demonstrated effect on ORNs involved in courtship and mating behavior1. Our procedure thus affords a new avenue to assay a host of low-volatility odorants for the study of insect olfaction and pheromone communication.

Electrophysiological Recording from Drosophila Trichoid Sensilla in Response to Odorants of Low Volatility

The overall goal of this protocol is to demonstrate how to present odorants of low volatility for single-sensillum recording from Drosophila olfactory receptor neurons that respond to long-chain cuticular pheromones.

电生理记录<em&gt;果蝇</em&gt; Trichoid Sensilla响应低挥发性气味

Insects rely on their sense of smell to guide a wide range of behaviors that are critical for their survival, such as food-seeking, predator avoidance, ...