作者要感谢澳大利亚国家健康和医学研究委员会和迪肯大学的支持。在迪肯大学感官实验室进行的这项工作是由国家健康和医学研究委员会资助（1043780）（RSJK）和澳大利亚园艺有限公司（BS12006）（RSJK）的支持。

下面的技术已被批准用于由迪肯大学人类研究伦理委员会使用。 1，人口统计学和人体测量学<ol><li>从参与者的记录人口统计信息，包括出生日期和性别。 </li><li>在基线（和其他研究指出，如果时间研究设计）采取的身高和体重的测量。确保参与者已经脱去鞋子，重型夹克或其他衣物，并从他们的口袋里取出任何重物。 <ol><li>用测距仪测量参与者的高度。记录测量到最近厘米。 </li><li>使用专用秤称量的参与者。记录重量为最接近的克。 </li><li>体重（kg）/身高2（米2）：使用公式计算BMI。由此看来，参加者按照标准定义的BMI值分类;健康18.5-25千克/米2，超重25-30公斤/米2或肥胖＆＃62，30千克/米2 27。 </li></ol></li></ol>口腔脂肪酸阈值评估2。生产样品<ol><li>使用非脂肪UHT奶作为基数脂肪酸味阈值评估。产品可以购买和贮存散装如有必要，将继续为未开封长达6个月，或直至该产品已达到其到期。准备2种类型的车辆：车辆添加了脂肪酸和控制车辆。为测试准备的解决方案的体积取决于参与者数目。下面的协议提供了典型的金额2人参加。 </li><li>准备一个基地乳溶液放置5％w来使用，在控制和脂肪酸车辆：V食品级阿拉伯胶（ 如 100克每2升的牛奶）到一个3升的玻璃烧杯中。如果需要，水合物胶在使用前（这将取决于口香糖制造商而异）。 </li><li>瓦特添加0.01％：V的EDTA的胶，以防止氧化（ 例如 ，2个每2升牛奶）00毫克。 </li><li>分配约1升每名参加非脂乳（ 例如 。，2参与者，用2升牛奶），倒入烧杯中。 </li><li>用实验室级混合器与emulsor屏幕，均质溶液在12,000 rpm离心2分钟。预留的解决方案。 </li><li>准备使用食品级的C18脂肪酸的解决方案：1。氧化可通过气相色谱法，如果必要进行评估。 </li><li>制备了一系列的脂肪酸车辆（UHT无脂牛奶）的13变体与C18的浓度增加：1（0.02，0.06，1，1.4，2，2.8，3.8，5，6.4，08 9.8，12，和20mM / L）。要做到这一点，标注250毫升的玻璃烧杯，每个浓度。 </li><li>添加5％的液体石蜡，以每个烧杯中（ 例如 ，每100毫升牛奶溶液5毫升石蜡）。 </li><li>根据C18：1浓度，添加C18适量的第1个至每烧杯（ 见表1）。 </li></ol><table border="1" cellpadding="0" cellspacing="0"&gt; <tbody><tr><td> C18：1浓度（MM） </td><td> 微升/ 100毫升 </td></tr><tr><td> 0.02 </td><td> 0.56 </td></tr><tr><td> 0.06 </td><td> 1.9 </td></tr><tr><td> 1 </td><td> 31.5 </td></tr><tr><td> 1.4 </td><td> 44.1 </td></tr><tr><td> 2 </td><td> 63.1 </td></tr><tr><td> 2.8 </td><td> 88.4 </td></tr><tr><td> 3.8 </td><td> 119.9 </td></tr><tr><td> 5 </td><td> 157.8 </td></tr><tr><td> 6.4 </td><td> 202 </td></tr><tr><td> 8 </td><td> 250 </td></tr><tr><td> 9.8 </td><td> 309 </td></tr><tr><td> 12 </td><td> 380 </td></tr><tr><td> 20 </td><td> 631.2 </td></tr></tbody></table> 表1实施例C18：每100毫升溶液1浓度增加C18的浓度（μL/ L）：（1）用于制备该系列的13乳剂口服脂肪酸苏氨酸eshold测试。 <ol start="10"><li>下面使用，填补了C18：1的容器与N 2，尽量减少氧化和存储低于4°C </li><li>添加的碱牛奶溶液各脂肪酸的烧杯中，以100ml的总体积。备用。 </li><li>利用剩余的碱溶液，准备控制车辆。在一个2升玻璃烧杯中，（在750毫升的最终体积如 35毫升液体石蜡）中添加液体石蜡剩余量的5％，而剩余的碱溶液一起和均匀化对于每100毫升的液体持续30秒。 </li><li>融汇控制车辆每100毫升30秒。这一步是先于脂肪酸的解决方案进行的，以防止污染与C18：1。 </li><li>匀化各脂肪酸的车辆中，具有最低浓度为每100毫升30秒开始。 </li><li>灭菌的均化之前和之后测试。 </li><li>由于同质化的过程可以提高solutio的温度NS，控制和C18的检查温度：1样品用温度计。为所有的样品在室温（20℃）。 </li><li>牛奶样品必须在同一天测试中新鲜制备的。味道在测试之前每个解决方案来评估的新鲜度和适用性。 注意：根据所需的体积，溶液的配制将需要至少60分钟。 </li></ol> 3，口服脂肪酸阈值测试<ol><li>确保参与者已经从进食或饮酒（包括咖啡，口香糖，漱口水等），至少1小时在测试之前忍住了。 </li><li>通过在红色的灯光与参加者戴着鼻夹进行测试尽量减少非味道线索。 </li><li>使用上升被迫选择三角形的程序，以确定口服脂肪酸阈值。标号30毫升塑料杯部与一个三位数的识别号码。为每个参与者提供了一组三个20毫升的解决方案以随机顺序;两个控制车辆和一个脂肪阿哲ð车辆与C18的最低浓度：1（0.02毫米）。 </li><li>以确定参与者的口服脂肪酸阈值时，指示所述参与者从左至右品尝每种溶液和吐出成片。要求学员无法下咽的样本。 </li><li>请参与者找出其中3个样品是&#39;奇怪&#39;或&#39;不同&#39;，如果他们不能确定，他们必须猜测（被迫选择）。 </li><li>有参加者各组样品后漱口用离子交换水。 </li><li>如果正确识别，提供参与者与第二组的3溶液（2控制及以随机顺序1脂肪酸溶液）具有相同的脂肪酸浓度。如果不正确地识别，提供参与者与第二组3个解决方案，但下一个最高浓度的C18：1（0.06毫米）。 </li><li>继续这个过程，直到参与者能够正确地识别出“奇”样本3倍行以相同的浓度。在其中它们能够正确地识别“奇”样本的浓度记录为参与者C18：1的检测阈值。参见图1为这一过程的图形表示。 </li><li>基于检测阈值，表征参与者敏感，或hyposensitive到C18：1。与以往文献线，过敏性个人可以检测到C18：1在浓度&lt;3.8毫米，而hyposensitive个人需要的浓度&gt; 3.8毫米。 注意：根据不正确的答案的数目，测试过程可以10-30分钟之间取来完成。 </li></ol><img alt="图1" fo:content-width="5in" fo:src="/files/ftp_upload/51236/51236fig1highres.jpg" src="/files/ftp_upload/51236/51236fig1.jpg" /> 用于测定脂肪酸检测t图1。递增被迫选择三角形的程序。hresholds参加者提供三种解决方案（两个控制解决方案和一个C18：在给定浓度1溶液），并要求确定了“奇怪的样本”。如果正确的话，参与者都获得了第二组具有相同的C18样品：1的浓度。如果不正确，则参与者设置有另一组样品与C18的较高浓度：1。此过程继续进行，直到3“奇”解决方案是在给定浓度正确识别。这一点被认为是个体的脂肪酸检测阈值“。 4，脂肪排名任务此任务涉及的参与者品尝四种样品瞬间蛋羹，各有不同的脂肪含量（0，2，6，和10％）和排名，以便他们认为升序脂肪浓度。 <ol><li>按照包装上的说明使用非脂肪速溶香草吉士粉准备1批蛋羹。混合2汤匙吉士宝wder，1汤匙白糖和2杯无脂牛奶在微波炉碗。如果建议的产品是不可用的，这可被类似的无脂 ​​肪速溶产品（ 例如 ，煮即成蛋奶）。 </li><li>采用高功率，在30秒的间隔用1400 W微波炉，总共约23分钟加热该混合物，或直到厚。取决于蛋羹的品牌和所用的微波的功率，这可能会有所不同。让蛋羹冷却。 </li><li>标签四500毫升厨房碗（或类似）与脂肪百分比。 </li><li>分割成蛋糊4个独立的100-G批次。 </li><li>添加0，2，6，和10％的植物油，以每碗要达到理想的脂肪含量（ 例如 ，在一个100克配料，加0毫升，分2毫升，6毫升，和10ml的植物油以达到各自的脂肪百分比）和结合。搅拌均匀每个样品以及确保所有成分都完全合并。 </li><li>标签四与随机THRE 30毫升的塑料杯部分电子数字。填补了部分杯20微克每吉士（1型每杯吉士）的。 </li><li>在测试之前冷藏的样品和服务冷（4℃）。 </li><li>进行下红灯测试，以减少视觉线索。 </li><li>有与会者品尝，吞咽和排名从感知最低4蛋羹，以脂肪含量最高，并获得得分超过了5根据他们的反应。 </li><li>得分为这个任务被示于表2。 注：大概准备时间吉士样品为30分钟。脂肪排名的任务应该不超过10分钟才能完成。 </li></ol><table align="left" border="1" cellpadding="0" cellspacing="0"><tbody><tr><td> 排名顺序 </td><td> 得分 </td></tr><tr><td> 0,2，6，10 </td><td> 5 </td></tr><tr><td> 2，0，6，10 </td><td> 4 </td></tr><tr><td> 0，2，10，6 </td><td> 3 </td></tr><tr><td> 0,6，2，10 </td><td> 2 </td></tr><tr><td> 1，6，10，2 </td><td> 1 </td></tr><tr><td> 6，0,2，10 </td><td> 1 </td></tr><tr><td> 2，10，06 0 </td><td> 1 </td></tr><tr><td> 2,6，0，10 </td><td> 0 </td></tr><tr><td> 6，2，10，0 </td><td> 0 </td></tr><tr><td> 0，10，2，6 </td><td> 0 </td></tr></tbody></table> 表2。脂肪排名得分任务，参加者给出4个样品吉士与0，2，6，或10％的脂肪补充说。参与者被要求来样订做排名从低到高脂肪含量和评分0〜5分（5分为最高）。 5，脂肪食物喜欢的<ol><li>准备市售食品的正常和低脂肪的选择小样本（5-20​​克）。食物包括定期和低脂版本：奶油奶酪（上一块饼干送达），巧克力慕斯，芝士，干饼干，花生酱浸送达了一块胡萝卜，蛋黄酱，沙拉博士埃辛（送达黄瓜切片），和酸奶。 </li><li>标记每个样品用随机三位数字的识别。 </li><li>随机顺序，以防止为了效果目前样品。 </li><li>指导参加单独品尝每个样品。食品摄入，但他们的愿望参与者可以吃多或尽可能少每个样品。 </li><li>有与会者评价他们多么喜欢或不喜欢每个样本。使用100毫米享乐广义震级衡量的喜好（GLMS; 见图2），从最强可以想象看不顺眼最强可以想象等。记录放置一个垂直线表示，如参加者或食物的厌恶的地步顺心。 </li></ol><img alt="图2" fo:content-width="5in" fo:src="/files/ftp_upload/51236/51236fig2highres.jpg" src="/files/ftp_upload/51236/51236fig2.jpg" /> 图2。享乐GLMS。享乐GLMS 30,31用于评估常规和低脂肪市售食物的喜好。与会者品尝和评价每个样品放置一个垂直线在哪个最能代表他们的像，或样品的厌恶的地步。 <ol start="6"><li>无味道的投入不会最小化这个任务，所以执行这个任务了普通光下没有参加戴鼻夹。 </li></ol> 6。舌摄影<ol><li>成立了相机和三脚架摄影。普通室内光线充足。 </li><li>为近距离摄影的相机设置到微距模式（或类似）。 </li><li>用打孔器建立在1.5厘米6毫米直径的圆X 1.5公分（或类似）的滤纸广场。标签纸与参与者的识别号码。 </li><li>在一个50毫升烧杯中，将结合用离子交换水蓝色食用着色剂以1:20的比例。每个参与者必须少量。 </li><li>倒30毫升富D级乙醇为50ml的烧杯镊子消毒。 </li><li>使用胶带，标记上测试台的侧一幅20厘米×30厘米的矩形（这个应该是正规的办公桌高度）， 如图3a所示 。 </li></ol><img alt="图3" fo:content-width="5in" fo:src="/files/ftp_upload/51236/51236fig3highres.jpg" src="/files/ftp_upload/51236/51236fig3.jpg" /> 图3 A）舌拍摄设置。演示前，舌头摄影B）舌摄影所需的表设置的。舌头摄影方法的示范<ol start="7"><li>有与会者放置在矩形的边角标明他们的胳膊肘，休息的下巴在他们的手掌和舒适地伸出自 ​​己的舌头，用嘴唇来稳定这个位置（ 图3b）。参与者必须保持在这个位置上的试验在持续时间克</li><li>使用矩形（1.5厘米×3cm）中的滤纸条，简要地干燥该舌的底部。 </li><li>蘸棉签插入食品着色/水溶液和传输少量染料到舌的前背表面，立即向右中线点和靠近顶端（参见图4）。干燥舌第二次用滤纸。 </li><li>无水乙醇消毒镊子用纸巾和使用镊子，将预标记1.5厘米2滤纸上在参与者的舌头，用6毫米的孔的蓝色食用着色剂（参见图4）。 </li><li>使用闪光灯，采取参与者的舌头三个数码照片。出于保密，确保只有参与者的嘴和舌头是可见的。 </li><li>从参与者的舌头已经再次经过消毒的食品级乙醇镊子取出1.5 平方厘米的滤纸。上传照片s到照片编辑软件，并具有缩放功能，计算所有可见的菌状乳头。 </li><li>从其他区分乳头菌状乳头为大蘑菇状，架空结构。他们没有考虑对染料溶液为强，因此出现在色轻得多。 注：舌拍摄时间不应超过10分钟才能完成。 </li></ol><img alt="图4" fo:content-width="5in" fo:src="/files/ftp_upload/51236/51236fig4highres.jpg" src="/files/ftp_upload/51236/51236fig4.jpg" /> 图4。量化菌状乳头密度。位置的6毫米面积为菌状乳头评估。使用摄影编辑软件，数值数字表示每个菌状乳头。

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作者什么都没有透露。

测定口服脂肪酸阈值，高脂肪食物的喜好，和舌乳头密度描述的技术进行了验证，并在一些已经发表的作品，近年来使用，我们建议口服脂肪酸的阈值评估，脂肪排名任务和油腻的食物的喜好一式两份，在一项研究中每个相关的时间点进行。目前尚无有关评估检测阈值32的最佳方法的一些讨论。特别地，使用溶液的组合物的实验室之间变化，一样的方法本身。具体地，所述脂肪酸在这个协议中使用，C18：1，我们相信这是一个通常代表和易于使用的脂肪酸，相对于其它脂肪酸，包括亚油酸（C18：2）和月桂酸（C12：0） ，先前已9使用。 C18：1是常用的食品供应和不同于C12实测值：0是液体在室温下，和对氧化比C18更耐：2 9。C18：1还显示出在多个测试会话提供可靠的数据，并且是高度相关的C18：2和C12：0 10。此外，C18：1已被广泛地研究整个相关文献，并因此进行比较更有帮助。 本文件和其他程序在其他实验室用内列出的协议之间差异的一个主要点是用于展示脂肪酸的刺激，并通过该检测阈值确定的系统的方法的车辆。该文献中使用的两种主要脂肪酸车辆是无脂乳10,17和水乳液6。 虽然两者都表现出疗效脂肪酸阈值确定，参与者可能更容易识别内乳脂肪的口感，即，它是不寻常的味道脂肪酸中的水，这可能会导致较低的水平外部有效性的用于利用研究水基。非脂乳为脂肪酸的化学感应车辆，不影响有效性。虽然这两种方法都尚未被直接比较在文献中，它是已知的脂肪酸是难溶于水33。由于牛奶为基础的解决方案脂肪酸的溶解度因此，该乳液既能保持时间更长，比水基溶液更均匀，虽然这尚未得到证实。当实现这种方法，必须注意的游离脂肪酸可以是天然存在于牛奶34，因此，该产品应使用以及其期满内，以防止游离脂肪酸（其随着年龄的发展）的增加，并与潜在的干扰是很重要的味阈值性能。成功制备解决方案取决于许多因素。首先，其中的“成分”的添加顺序是必要的。车辆准备步骤应该仔细按照这些早期的概述，以确保适当的车辆组成第二稳定的乳液。其次，温度必须控制对。每个样品必须向与会者介绍了在室温，确保学员不检测的“奇样本&#39;由于种种因素，比&#39;味道&#39;等。最后，所有的样品必须被正确地均化的时间所建议的时间。而脂肪酸的乳液和无脂乳比如果水是用作更有效的，但仍然在样品中乳液分离的机会。 在口服脂肪酸阈值确定所使用的具体测试方法也必须加以考虑。两个感觉为基础的方法已在文献中被广泛描述的;一个是被迫上升三角形选择程序和替代，楼梯方法35。升被迫选择三角形的方法是一个既定的方法味阈值测定，可以认为有用的有几个原因，包括事实，不像楼梯方法，升方法开始于C18的最低浓度：1（0.02毫摩尔）和增加直到参与者能够检测在溶液9脂肪酸的存在。相反，楼梯方法包括增加或从预定的中点11降低脂肪酸的浓度。然而，在上述阈值的点开始阈值确定可能造成的损害的反应品尝那些能力脱敏。此外，递增方法具有随机的机会影响的结果（3.7％）与阶梯法（11.1％）11相比具有较低的概率。因此，我们建议升强制选择三角法，并结合非脂肪乳作为口味测试的车辆似乎是准确地确定口服阈值的有效手段。 食品的接受或喜欢的测试是感官研究中所执行的更简单的评估1，因此有几个问题吨帽子往往出现。然而，使用喜好规模类型是一个重要的焦点。在这种情况下，一个享乐GLMS是最有效的，因为它具有良好的分辨能力，并且容易为参与者使用36。享乐GLMS的结束点都标注了描述“最强可以想象厌恶”和“最强可以想象像&#39;和参与者评估顺心反对一切享乐体验，而不是单纯的食物30,31。这是有效地控制用于通过标准的9点量表产生天花板效应，因为所有的经验被认为和比较。此外，享乐GLMS更能够表现出更大的个体差异，为规模更广泛的36。食品验收测试本身所提出的食物是有限的，在每类食物，我们目前只有两个选择。进一步的研究可能包括几个品牌或类型每种食物，各有不同的脂肪含量，或者是专门制造的产品，其中的脂肪内容可被控制，并且是唯一的变量。需要注意的是所有数据的解释必须谨慎进行是很重要的。虽然喜好，偏好和摄入之间的潜在联系是合理的和有趣的，是在实验室环境中产生的结果，并有可能限制这些研究结果是否适用于真实世界的情况。 通过舌头摄影评估乳头密度是一个比较困难的过程，与必须采取以产生适当和适用结果的具体步骤。特别是，它以确定正确的乳头类型是重要的。三种类型的味觉乳头的是对人舌可见;菌状，叶状和circumvallate 4。菌状乳头但是，我们可以很容易地分辨为蘑菇形结构26，而且一般是在敏感性评估记录的乳头。菌状乳头往往集中在5-60的范围内每6毫米面积37（取决于灵敏度），虽然已经有研究表明，有些人可能有230乳头向上同一地区21。使用相机的类型是基本获得相应的结果，并且可以解释这种差异。此前在这方面的使用数码摄影，电视显微镜是黄金标准标识和记录乳突密度。然而，已经确定，识别相同的水平，可以利用适当的数字照相机26。此外，数码摄影仅需几分钟，其中电视显微镜可能需要长达一个小时26。不仅如此，但数码摄影必须远成本更低，并且更便于携带，这可以是用于与各种参与者组26使用有帮助的可能性。最后，虽然我们的目标是衡量菌状乳头密度与口腔脂肪酸检测协会，我们也建议口味阈值的t他5原型口味也可以并行执行。鉴于与乳突密度和味觉功能以前的联动，这个可以作为一个额外的“检查措施”，这可能会增加完整性的数据，尤其是考虑到这是一个新的研究领域。 口服化学感受研究领域，特别是关于脂肪酸，是一个新兴的1，因此，重要的是所有的研究被执行以高标准，优选与使用的一致的协议，以允许直接比较。

过多的膳食脂肪的消耗是一个潜在的贡献者体重增加1-3和肥胖已成为现代全球性的流行病。研究表明，较高水平的脂肪摄入量，尤其是作为一个自由采食饮食的一部分，可能与较高的BMI 2,3有关，但影响食物中脂肪消费和偏好的因素并不清晰。搜寻在该背后消耗脂肪的机制，因此具有明显的目标，特别令人感兴趣的是口服机构负责脂肪检测，通常被称为“脂肪酸味道&#39;2。 从进化的角度来看，味觉系统想必担任消化系统的把关，指导能源密集的营养成分和驱逐潜在的有毒化合物4的消耗。味觉是通过其分布于三个类型的舌头的专门味觉受体细胞引发乳头;菌状，叶状，和circumvallate乳头，它可以分别含有高达几百味蕾5。除了 ​​被广泛接受的5典型的口味（甜，咸，酸，苦和鲜味），它并不完全令人惊讶的是一直存在的口头机制用于 ​​检测脂肪，或者更可能的分解产物脂肪酸6建议。 以前的研究一致表明脂肪酸能在口腔中超过一定浓度范围7-11被检测到，尽管它不是传统意义上的&#39;味道&#39;，因为它有一个与之关联的单一明显感知质量（ 即 。甜）12。从我们的实验室工作突出减值脂肪酸化学感受功能的影响，即对体重和膳食脂肪的消耗。那些谁是不太能够检测脂肪酸（hyposensitive）似乎有较高的身体质量指数（BMI）和消耗更多能源9，而与口腔脂肪酸的敏感性和膳食脂肪消耗的关系也被观察到;也就是说，脂肪酸hyposensitive个人表现出要消耗更多的动物脂肪，包括肉类，高脂肪的乳制品，脂肪和传播所有这些都被牵连贡献者体重增加13。此外，个人谁是脂肪酸更敏感似乎是更好的装备在具有不同脂肪含量的9个样品之间的区别。而其他研究小组都未能找到类似的协会10,14,15，这个不断增长的研究领域仍然耐人寻味。 在脂肪酸化学感受这些个体差异似乎在一定程度上受到环境因素，包括饮食进行调制。习惯性脂肪的摄入量已受损的口腔脂肪酸化学感受，因此，一个高度偏爱有关，而消费增加饮食中的脂肪16。除了 ​​味觉适应，胃肠道（GI）也出现顺应这种变化，脂肪摄入量和17胃肠受损脂肪酸的敏感性可能无法产生适当的饱腹感信号的劝阻多余的能量消耗18的反应受到牵连。 除了 ​​环境因素，脂肪酸化学感受也可以通过个体之间的遗传或生理上的差异，包括菌状乳头密度对一个人的舌头19的浓度（大概是味觉受体）所决定的。高密度的菌状舌乳头已被链接到提高口腔灵敏度为众多的口服检测化合物，包括6 - 正丙基硫氧嘧啶（PROP）20，糖21，盐22，而其他人也注意到，奶油感知23的关联。 PROP supertasters（谁presumaBLY有许多菌状乳头以上）能够从低脂肪的沙拉酱24分清 ​​高脂肪，并能比非品酒师23,25更加准确地判别脂肪含量的奶制品和奶油感。在此阶段然而，菌状乳头密度和口服脂肪酸“味道”的检测之间的关系是未知的。 在人类口腔脂肪酸化学感受研究的基础是各种感官技术的应用。识别个体差异在口服脂肪酸的检测是一个主要的焦点，并在很大程度上取决于脂肪酸的检测阈值的确定，即，在该脂肪酸的最低点是能够在溶液9被检测到。尽管所使用的特定的测试方法，并刺激整个车辆的文献和研究团体之间变化，典型的过程包括提出一个参与者提供了一套乳化脂肪酸和对照组（没有的f阿蒂酸）解决方案，并确定哪些是“奇”的样本。在这里，我们提出了使用乳化牛奶的解决方案和一个上升被迫选择三角形的过程阈值决定10建立的，可靠的和可重复的方法。 在多大程度上口头脂肪酸敏感性影响饮食，即食用脂肪类食物，并认为脂肪食物的能力是兴趣也是在这里，我们对另外两个成熟的技术也报告，以进一步扩大我们的脂肪酸化学感受的理解。高脂肪食物的喜好可以通过为个人提供的市售食品样品，既具有常规和低脂肪的选择谁被要求来表示每个16的喜好来确定。在问候脂肪感知，一个胖胖的排名工作已经制定了我们的实验室，目的是评估一个人的检测脂肪蛋羹，一个典型的食品基质16的能力。为了评估genetic或个人之间的生理差异，通常使用的舌摄影方法包括染色，照相，定量菌状乳头26。虽然使用这种技术中脂肪酸的研究正处于起步阶段，应用越来越普遍，尤其是在这两个精益和超重/肥胖人群可能有助于确定消耗多余脂肪的内在原因。

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			<td height="42" style="height:42px;width:216px;">Name of Material/ Equipment</td>
			<td style="width:104px;">Company</td>
			<td style="width:119px;">Catalog Number</td>
			<td style="width:471px;">Comments/Description</td>
		</tr>
		<tr height="63">
			<td height="63" style="height:63px;width:216px;">Gum Arabic TIC Pretested PRE-HYDRATED FT Powder</td>
			<td style="width:104px;">Alchemy Agencies Ltd. NZ</td>
			<td style="width:119px;">CFR# 21 CFR 184.1330</td>
			<td>Food grade agrigum</td>
		</tr>
		<tr height="63">
			<td height="63" style="height:63px;width:216px;">EDTA (Ethylenediaminetetraacetic acid)</td>
			<td style="width:104px;">Merck</td>
			<td style="width:119px;">1.08418 0250</td>
			<td>Disodium salt dehydrate</td>
		</tr>
		<tr height="63">
			<td height="63" style="height:63px;width:216px;">L4RT Homogenizer</td>
			<td style="width:104px;">Silverson Longmedow, MA</td>
			<td style="width:119px;">L4RT</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Liquid Paraffin</td>
			<td style="width:104px;">Fauldings</td>
			<td style="width:119px;"></td>
			<td>No catalog number as liquid paraffin is a regular consumable product</td>
		</tr>
		<tr height="63">
			<td height="63" style="height:63px;width:216px;">Nikon AF-S VR Micro Nikkor 105mm f/2.8G IF-ED camera lens</td>
			<td style="width:104px;">Nikon</td>
			<td align="right" style="width:119px;">2160</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">SLIK Sprint Pro II tripod</td>
			<td style="width:104px;">Slik Corporation</td>
			<td>611-849</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Nikon D90 Digital Camera with LCD Protector</td>
			<td style="width:104px;">Nikon</td>
			<td style="width:119px;">BM-10</td>
			<td></td>
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		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Nitrogen</td>
			<td style="width:104px;"></td>
			<td style="width:119px;"></td>
			<td></td>
		</tr>
		<tr height="63">
			<td height="63" style="height:63px;width:216px;">Tanita Body Scan Composition Monitor Scales</td>
			<td style="width:104px;">Tanita, Cloverdale, WA, Australia</td>
			<td style="width:119px;">BC-551</td>
			<td></td>
		</tr>
		<tr height="84">
			<td height="84" style="height:84px;width:216px;">Seca Stadiometer</td>
			<td style="width:104px;">Medshop Australia, Fairfield, VIC, Australia</td>
			<td style="width:119px;">MED435</td>
			<td></td>
		</tr>
	</tbody>
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measuring oral fatty acid thresholds, fat perception, fatty food liking, and papillae density in humans

从一些实验室新的证据表明，人类必须确定脂肪酸在口腔内，据推测是通过安置在味觉细胞脂肪酸受体的能力。以前的研究已经表明，一个人的口头敏感性脂肪酸，特别是油酸（C18：1）与身体质量指数（BMI）进行相关，饮食中的脂肪量，并确定食品中脂肪的能力。我们已经开发出一种可靠的和可再现的方法来评估的脂肪酸口服化学感受，用牛奶和C18：1的乳液，与上升的强迫选择三角形程序一起。与此同时，食品矩阵已经发展到评估一个人的感知脂肪，除了一个简单的方法来评估脂肪的食物喜好的能力。作为补充措施舌摄影是用来评估乳头密度，具有较高的密度常常被与增加的味道敏感性相关。

上文详述的方法是重要的，因为一些新的证据已表明，在口腔和胃肠道障碍脂肪酸化学感受可以被增加，BMI和肥胖17的发展相关联。几个研究已经使用了所述协议来调查口服脂肪酸的检测以及最近的出版物已经表明该方法是可靠和可重复的10。已使用这种方法的研究已经能够通过识别该点的参与者都能够检测牛奶样品9之间的差别可靠地确定个人的口腔脂肪酸的检测阈值。 。经过三年的测试阶段，利用这个协议中，斯图尔特等[9]发现，对于C18的平均检测阈值：1为2.2±0.1，与检测阈值范围为1-6.4毫米（ 见图5）。最近，我们已经建立了C18：1的检测阈值范围从0.26-12毫米，平均（2.64±0.7毫米）10。这些结果支持了脂肪酸可以在口腔中被检测到，并且该标识的灵敏度〜18的个体差异的概念：1存在。基于这些结果，我们可以划分为个人或过敏性hyposensitive到C18：1。过敏性个人都能够正确识别的C18：1 &lt;3.8毫米，而hyposensitive科目要求的浓度&gt; 3.8毫米。我们实验室的研究发现口服灵敏度C18：1 hyposensitive个人消耗更多的饱和动物脂肪，并有较高的BMI 13：1，与食物中的脂肪消耗和体重指数（ 图6），其中C18相关有趣的是，在由Stewart和Keast的16进行了一项研究，发现食用低脂肪饮食导致对C18的敏感性增加：1为精简和超重参与者（ 图7）。然而，这项研究还发现，当参与者consu<html> MED高脂肪的饮食，瘦了个体敏感性降低C18：1，而超重的人有味道的敏感性没有变化（ 图8）。这表明，习惯性地消耗高脂肪食物，这是更容易为超重个体，可能导致减毒脂肪酸化学感受28。然而，由于有在精益和超重参与者之间的基线灵敏度无显着差异，这些结果可能表明，精益人只是更容易受到有关脂肪的摄入饮食结构的改变。这也可能表明，它是特定干预（高与低脂肪）的存在下，可能已影响的结果，而不是惯常的饮食，这可能会或可能不会有不同的饮食干预。尽管如此，这项研究表明，有关于脂肪酸味道的敏感性，这需要进一步调查瘦肉和超重的人之间有一些根本性的分歧。 ontent“FO：保持together.within页=”总是“&gt; <img alt="图5" fo:content-width="5in" fo:src="/files/ftp_upload/51236/51236fig5highres.jpg" src="/files/ftp_upload/51236/51236fig5.jpg" /> 图5 C18：1，味检测阈值已标记的变异已在敏感的C18被证明：（1）与参加者能够探测到C18：1在一系列浓度（1毫米- 6.4毫米）的。 <img alt="图6" fo:content-width="5in" fo:src="/files/ftp_upload/51236/51236fig6highres.jpg" src="/files/ftp_upload/51236/51236fig6.jpg" /> 图6 C18：1，味检测阈值并与BMI的关联 ，以检测C18的能力之间的关联：1和身体成分已被证明，由此那些具有较高的检测阈值（hyposensitive个人）有显著更高的BMI值（P = 0.002 ，R 2 = 0.467）。 图7 C18：1，检 ​​测阈值以下的低脂肪饮食经过低脂肪饮食，C18 4周为1消费：1检测阈值增加为精简和肥胖者。 <img alt="图8" fo:content-width="5in" fo:src="/files/ftp_upload/51236/51236fig8highres.jpg" src="/files/ftp_upload/51236/51236fig8.jpg" /> 图8 C18：1，检 ​​测阈值以下的高脂肪食物经过高脂饮食4周为1消费，精益显示的个体敏感性降低C18：1（P = 0.006），而超重的人没有表现出变化（P = 0.609） 。 同样的饮食对检测阈值脂肪酸的影响，有研究suggeST食物的喜好可能是塑料和曝光改变。例如，似乎高脂肪饮食增加偏好较高脂肪的产品，以产生下列一种低脂肪饮食16的消耗量相反。然而，这些变化还没有在文献中是一致的。看来，更改的偏好是由时间的个体一直秉承高或低脂肪饮食的长度调节。具体来说，马特斯29发现参与者的饮食偏好显著变化12周减少脂肪饮食后，而斯图尔特和Keast的16只发现零星的边际变化四周内就类似的饮食后。食用高脂肪的饮食改变参与者的偏好酸奶，有偏好低脂酸奶增加，相反到预期的结果（基线（BL）：19.44±5.73，第4周（WK 4）：21.94±5.21，P = 0.046）。另外，有4周的低脂肪饮食后，偏好低脂肪奶油在所有的p增加articipants（BL：6.23±4.26，WK 4：7.32±3.04，P = 0.046）。为低脂酸奶的喜好增加瘦肉参与者只（BL：2.51±3.26，WK 4：3.68±4.94，P = 0.07），而偏好低脂慕斯下降为所有参与者（P = 0.01）。 检测脂肪食物的能力是通过要求参与者品尝，并享有一系列蛋羹具有不同脂肪含量的评估。脂肪的感知是基于参加者如何以及能够排在样品标识。脂肪的感知已经知道，改变饮食，例如，下面的低脂肪饮食，导致改善的正确识别和排名的脂肪程度蛋奶每个样品11在参与者的表现。此外，它似乎有一个协会敏感性之间的C18：1和鉴定脂肪含量4和排名。事实上，谁是过敏的18个人：1表现显著好Øn中的脂肪排名任务（4.3±0.6）相比，hyposensitive个人（2.3±0.1，P = 0.02）（分数满分为5）9。这表明，个人谁是脂肪酸更敏感也是在四个不同脂肪含量的牛奶蛋糊区分好。虽然有一种趋势，性能消耗低脂肪饮食四周后改善，这不是一个显著的变化（BL：1.3±0.3，WK 4：2±0.3，P = 0.077）16。 乳头密度即乳头的数量（并且因此味蕾）存在于舌头个体之间变化，并且是指示味功能的。较高的菌状乳头密度已与品味高度敏感性，对化合物，包括蔗糖21和苦味物质PROP 20。舌乳头密度，由舌头摄影确定，不同学科之间相当。例如，张&lt;EM&gt;等21发现有参加者之间显著的个体差异，范围从7.07±0.35/cm 2至233.43±0.00/cm 2浓度（数据是对一个参与者），而其他26人发现，平均菌状乳头浓度为156.00±5.86/cm 2。此外，已经发现，乳头可出现显著不同的方式在个人之间的结构，具有变化的高度，宽度和形状，21虽然关于这些差异可能造成的影响有限的证据。鉴于以往的调查结果联系起来有品味的灵敏度乳头数，它是合理的，类似的关系也可以口服脂肪酸的敏感性，使那些谁更口头脂肪酸的敏感可能有味觉乳头的密度更高，因此一些存在较高口服脂肪酸受体。虽然这个协会是尚未建立，它预先选择的窗口NTS一个新的研究领域可能有助于指导牵连脂肪的过度消费的基本机制。

Watch this Scientific Journal Video about Measuring Oral Fatty Acid Thresholds, Fat Perception, Fatty Food Liking, and Papillae Density in Humans at JoVE.com

Measuring Oral Fatty Acid Thresholds, Fat Perception, Fatty Food Liking, and Papillae Density in Humans | Neuroscience | Video

Measuring Oral Fatty Acid Thresholds, Fat Perception, Fatty Food Liking, and Papillae Density in Humans

脂肪酸，与饮食和高脂肪食物的喜好协会口服化学感受可能使涉及与肥胖的发展，以及为什么改变饮食可能难以对许多人机制的鉴定。

测量口腔脂肪酸阈值，感知脂肪，脂肪食物喜欢的，和乳头密度的人类

Emerging evidence from a number of laboratories indicates that humans have the ability to identify fatty acids in the oral cavity, presumably via fatty ...

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19.8K 次观看.  Deakin University. 脂肪酸，与饮食和高脂肪食物的喜好协会口服化学感受可能使涉及与肥胖的发展，以及为什么改变饮食可能难以对许多人机制的鉴定。 

视频: Measuring Oral Fatty Acid Thresholds, Fat Perception, Fatty Food Liking, and Papillae Density in Humans

Using Affordable LED Arrays for Photo-Stimulation of Neurons

Standard slice electrophysiology has allowed researchers to probe individual components of neural circuitry by recording electrical responses of single cells in response to electrical or pharmacological manipulations1,2. With the invention of methods to optically control genetically targeted neurons (optogenetics), researchers now have an unprecedented level of control over specific groups of neurons in the standard slice preparation. In particular, photosensitive channelrhodopsin-2 (ChR2) allows researchers to activate neurons with light3,4. By combining careful calibration of LED-based photostimulation of ChR2 with standard slice electrophysiology, we are able to probe with greater detail the role of adult-born interneurons in the olfactory bulb, the first central relay of the olfactory system. Using viral expression of ChR2-YFP specifically in adult-born neurons, we can selectively control young adult-born neurons in a milieu of older and mature neurons. Our optical control uses a simple and inexpensive LED system, and we show how this system can be calibrated to understand how much light is needed to evoke spiking activity in single neurons. Hence, brief flashes of blue light can remotely control the firing pattern of ChR2-transduced newborn cells.

Adult-born neurons expressing ChR2 can be manipulated in slice electrophysiological preparations in order to examine their contribution towards the function of olfactory neural circuits.

使用光刺激神经元的负担得起的LED阵列

Standard slice electrophysiology has allowed researchers to probe individual components of neural circuitry by recording electrical responses of single ...

科研

神经科学

Selective Viral Transduction of Adult-born Olfactory Neurons for Chronic in vivo Optogenetic Stimulation

Local interneurons are continuously regenerated in the olfactory bulb of adult rodents1-3. In this process, called adult neurogenesis, neural stem cells in the walls of the lateral ventricle give rise to neuroblasts that migrate for several millimeters along the rostral migratory stream (RMS) to reach and incorporate into the olfactory bulb. To study the different steps and the impact of adult-born neuron integration into preexisting olfactory circuits, it is necessary to selectively label and manipulate the activity of this specific population of neurons. The recent development of optogenetic technologies offers the opportunity to use light to precisely activate this specific cohort of neurons without affecting surrounding neurons4,5. Here, we present a series of procedures to virally express Channelrhodopsin2(ChR2)-YFP in a temporally restricted cohort of neuroblasts in the RMS before they reach the olfactory bulb and become adult-born neurons. In addition, we show how to implant and calibrate a miniature LED for chronic in vivo stimulation of ChR2-expressing neurons.

Selective Viral Transduction of Adult-born Olfactory Neurons for Chronic in vivo Optogenetic Stimulation

Adult-born neurons of the olfactory bulb can be optogenetically controlled using Channelrhodopsin2-expressing lentiviral injection in the rostral migratory stream and chronic photostimulation with an implanted miniature LED.

慢性成人出生的嗅觉神经元的选择性病毒转导在体内 Optogenetic刺激

Local interneurons are continuously regenerated in the olfactory bulb of adult rodents1-3. In this process, called adult neurogenesis, neural stem cells ...

MPI CyberMotion Simulator: Implementation of a Novel Motion Simulator to Investigate Multisensory Path Integration in Three Dimensions

Path integration is a process in which self-motion is integrated over time to obtain an estimate of one's current position relative to a starting point 1. Humans can do path integration based exclusively on visual 2-3, auditory 4, or inertial cues 5. However, with multiple cues present, inertial cues - particularly kinaesthetic - seem to dominate 6-7. In the absence of vision, humans tend to overestimate short distances (&lt;5 m) and turning angles (&lt;30&deg;), but underestimate longer ones 5. Movement through physical space therefore does not seem to be accurately represented by the brain.
Extensive work has been done on evaluating path integration in the horizontal plane, but little is known about vertical movement (see 3 for virtual movement from vision alone). One reason for this is that traditional motion simulators have a small range of motion restricted mainly to the horizontal plane. Here we take advantage of a motion simulator 8-9 with a large range of motion to assess whether path integration is similar between horizontal and vertical planes. The relative contributions of inertial and visual cues for path navigation were also assessed.
16 observers sat upright in a seat mounted to the flange of a modified KUKA anthropomorphic robot arm. Sensory information was manipulated by providing visual (optic flow, limited lifetime star field), vestibular-kinaesthetic (passive self motion with eyes closed), or visual and vestibular-kinaesthetic motion cues. Movement trajectories in the horizontal, sagittal and frontal planes consisted of two segment lengths (1st: 0.4 m, 2nd: 1 m; &plusmn;0.24 m/s2 peak acceleration). The angle of the two segments was either 45&deg; or 90&deg;. Observers pointed back to their origin by moving an arrow that was superimposed on an avatar presented on the screen.
Observers were more likely to underestimate angle size for movement in the horizontal plane compared to the vertical planes. In the frontal plane observers were more likely to overestimate angle size while there was no such bias in the sagittal plane. Finally, observers responded slower when answering based on vestibular-kinaesthetic information alone. Human path integration based on vestibular-kinaesthetic information alone thus takes longer than when visual information is present. That pointing is consistent with underestimating and overestimating the angle one has moved through in the horizontal and vertical planes respectively, suggests that the neural representation of self-motion through space is non-symmetrical which may relate to the fact that humans experience movement mostly within the horizontal plane.

An efficient way to gain insight into how humans navigate themselves in three dimensions is described. The method takes advantage of a motion simulator capable of moving observers in ways unattainable by traditional simulators. Results confirm that movement in the horizontal plane is underestimated, while vertical movement is overestimated.

MPI Cyber​​Motion模拟器：一种新型的运动模拟器的执行情况调查三个维度的多感官的路径积分

Path integration is a process in which self-motion is integrated over time to obtain an estimate of one's current position relative to a starting point 1. ...

Isolation and Culture of Human Fungiform Taste Papillae Cells

Taste cells are highly specialized, with unique histological, molecular and physiological characteristics that permit detection of a wide range of simple stimuli and complex chemical molecules contained in foods. In human, individual fungiform papillae contain from zero to as many as 20 taste buds. There is no established protocol for culturing human taste cells, although the ability to maintain taste papillae cells in culture for multiple cell cycles would be of considerable utility for characterizing the molecular, regenerative, and functional properties of these unique sensory cells. Earlier studies of taste cells have been done using freshly isolated cells in primary culture, explant cultures from rodents, or semi-intact taste buds in tissue slices1,2,3,4. Although each of these preparations has advantages, the development of long-term cultures would have provided significant benefits, particularly for studies of taste cell proliferation and differentiation. Several groups, including ours, have been interested in the development and establishment of taste cell culture models. Most attempts to culture taste cells have reported limited viability, with cells typically not lasting beyond 3-5 d5,6,7,8. We recently reported on a successful method for the extended culture of rodent taste cells9. We here report for the first time the establishment of an in vitro culture system for isolated human fungiform taste papillae cells. Cells from human fungiform papillae obtained by biopsy were successfully maintained in culture for more than eight passages (12 months) without loss of viability. Cells displayed many molecular and physiological features characteristic of mature taste cells. Gustducin and phospholipase C &beta;2, (PLC-&beta;2) mRNA were detected in many cells by reverse transcriptase-polymerase chain reaction and confirmed by sequencing. Immunocytochemistry analysis demonstrated the presence of gustducin and PLC-&beta;2 expression in cultured taste cells. Cultured human fungiform cells also exhibited increases in intracellular calcium in response to appropriate concentrations of several taste stimuli indicating that taste receptors and at least some of the signalling pathways were present. These results sufficient indicate that taste cells from adult humans can be generated and maintained for at least eight passages. Many of the cells retain physiological and biochemical characteristics of acutely isolated cells from the adult taste epithelium to support their use as a model taste system. This system will enable further studies of the processes involved in proliferation, differentiation and function of mammalian taste receptor cells in an in vitro preparation.
Human fungiform taste papillae used for establishing human fungiform cell culture were donated for research following proper informed consent under research protocols that were reviewed and approved by the IRB committee. The protocol (#0934) was approved by Schulman Associates Institutional Review Board Inc., Cincinnati, OH. Written protocol below is based on published parameters reported by Ozdener et al. 201110.

We aimed to develop a reproducible protocol for isolating and maintaining long-term cultures of human fungiform taste papillae cells. Cells from human fungiform papillae obtained by biopsy were successfully maintained in culture for more than eight passages (12 months) without loss of viability.

人力蕈状味觉​​乳头细胞的分离和培养

Taste cells are highly specialized, with unique histological, molecular and physiological characteristics that permit detection of a wide range of simple ...

Progressive-ratio Responding for Palatable High-fat and High-sugar Food in Mice

Foods that are rich in fat and sugar significantly contribute to over-eating and escalating rates of obesity. The consumption of palatable foods can produce a rewarding effect that strengthens action-outcome associations and reinforces future behavior directed at obtaining these foods. Increasing evidence that the rewarding effects of energy-dense foods play a profound role in overeating and the development of obesity has heightened interest in studying the genes, molecules and neural circuitry that modulate food reward1,2. The rewarding impact of different stimuli can be studied by measuring the willingness to work to obtain them, such as in operant conditioning tasks3. Operant models of food reward measure acquired and voluntary behavioral responses that are directed at obtaining food. A commonly used measure of reward strength is an operant procedure known as the progressive ratio (PR) schedule of reinforcement.4,5 In the PR task, the subject is required to make an increasing number of operant responses for each successive reward. The pioneering study of Hodos (1961) demonstrated that the number of responses made to obtain the last reward, termed the breakpoint, serves as an index of reward strength4. While operant procedures that measure changes in response rate alone cannot separate changes in reward strength from alterations in performance capacity, the breakpoint derived from the PR schedule is a well-validated measure of the rewarding effects of food. The PR task has been used extensively to assess the rewarding impact of drugs of abuse and food in rats (e.g.,6-8), but to a lesser extent in mice9. The increased use of genetically engineered mice and diet-induced obese mouse models has heightened demands for behavioral measures of food reward in mice. In the present article we detail the materials and procedures used to train mice to respond (lever-press) for a high-fat and high-sugar food pellets on a PR schedule of reinforcement. We show that breakpoint response thresholds increase following acute food deprivation and decrease with peripheral administration of the anorectic hormone leptin and thereby validate the use of this food-operant paradigm in mice.

The present report details the protocol employed to measure the rewarding effects of high-fat food in mice using a progressive ratio operant conditioning task.

累进比率响应小鼠可口的高脂肪和高糖食品

Foods that are rich in fat and sugar significantly contribute to over-eating and escalating rates of obesity. The consumption of palatable foods can ...

RNAi-mediated Double Gene Knockdown and Gustatory Perception Measurement in Honey Bees (Apis mellifera)

This video demonstrates novel techniques of RNA interference (RNAi) which downregulate two genes simultaneously in honey bees using double-stranded RNA (dsRNA) injections. It also presents a protocol of proboscis extension response (PER) assay for measuring gustatory perception.	
	RNAi-mediated gene knockdown is an effective technique downregulating target gene expression. This technique is usually used for single gene manipulation, but it has limitations to detect interactions and joint effects between genes. In the first part of this video, we present two strategies to simultaneously knock down two genes (called double gene knockdown). We show both strategies are able to effectively suppress two genes, vitellogenin (vg) and ultraspiracle (usp), which are in a regulatory feedback loop. This double gene knockdown approach can be used to dissect interrelationships between genes and can be readily applied in different insect species.	
	The second part of this video is a demonstration of proboscis extension response (PER) assay in honey bees after the treatment of double gene knockdown. The PER assay is a standard test for measuring gustatory perception in honey bees, which is a key predictor for how fast a honey bee's behavioral maturation is. Greater gustatory perception of nest bees indicates increased behavioral development which is often associated with an earlier age at onset of foraging and foraging specialization in pollen. In addition, PER assay can be applied to identify metabolic states of satiation or hunger in honey bees. Finally, PER assay combined with pairing different odor stimuli for conditioning the bees is also widely used for learning and memory studies in honey bees.

RNAi-mediated Double Gene Knockdown and Gustatory Perception Measurement in Honey Bees Apis mellifera

In this protocol, we describe two strategies that simultaneously suppress two genes (double gene knockdown) in honey bees. Then we present how to use the proboscis extension response (PER) assay to study the effect of double gene knockdown on honey bee gustatory perception.

RNAi介导的双基因敲除和味觉感知测量蜜蜂（<em&gt;蜜蜂</em&gt;）

This video demonstrates novel techniques of RNA  interference (RNAi) which downregulate two genes simultaneously in honey bees  using double-stranded RNA ...

The Neuromuscular Junction: Measuring Synapse Size, Fragmentation and Changes in Synaptic Protein Density Using Confocal Fluorescence Microscopy

The neuromuscular junction (NMJ) is the large, cholinergic relay synapse through which mammalian motor neurons control voluntary muscle contraction. Structural changes at the NMJ can result in neurotransmission failure, resulting in weakness, atrophy and even death of the muscle fiber. Many studies have investigated how genetic modifications or disease can alter the structure of the mouse NMJ. Unfortunately, it can be difficult to directly compare findings from these studies because they often employed different parameters and analytical methods. Three protocols are described here. The first uses maximum intensity projection confocal images to measure the area of acetylcholine receptor (AChR)-rich postsynaptic membrane domains at the endplate and the area of synaptic vesicle staining in the overlying presynaptic nerve terminal. The second protocol compares the relative intensities of immunostaining for synaptic proteins in the postsynaptic membrane. The third protocol uses Fluorescence Resonance Energy Transfer (FRET) to detect changes in the packing of postsynaptic AChRs at the endplate. The protocols have been developed and refined over a series of studies. Factors that influence the quality and consistency of results are discussed and normative data are provided for NMJs in healthy young adult mice.

The neuromuscular junction (NMJ) is altered in a variety of conditions that can sometimes culminate in synaptic failure. This report describes fluorescence microscope-based methods to quantify such structural changes.

神经肌肉接头处：测量突触大小，碎片和改变突触蛋白密度利用共聚焦荧光显微镜

The neuromuscular junction (NMJ) is the large, cholinergic relay synapse through which mammalian motor neurons control voluntary muscle contraction. ...

Non-invasive Assessment of Changes in Corticomotoneuronal Transmission in Humans

The corticospinal pathway is the major pathway connecting the brain with the muscles and is therefore highly relevant for movement control and motor learning. There exists a number of noninvasive electrophysiological methods investigating the excitability and plasticity of this pathway. However, most methods are based on quantification of compound potentials and neglect that the corticospinal pathway consists of many different connections that are more or less direct. Here, we present a method that allows testing excitability of different fractions of the corticospinal transmission. This so called H-reflex conditioning technique allows one to assess excitability of the fastest (monosynaptic) and also polysynaptic corticospinal pathways. Furthermore, by using two different stimulation sites, the motor cortex and the cervicomedullary junction, it allows not only differentiation between cortical and spinal effects but also assessment of transmission at the corticomotoneural synapse. In this manuscript, we describe how this method can be used to assess corticomotoneural transmission after low-frequency repetitive transcranial magnetic stimulation, a method that was previously shown to reduce excitability of cortical cells. Here we demonstrate that not only cortical cells are affected by this repetitive stimulation but also transmission at the corticomotoneuronal synapse at the spinal level. This finding is important for the understanding of basic mechanisms and sites of neuroplasticity. Besides investigation of basic mechanisms, the H-reflex conditioning technique may be applied to test changes in corticospinal transmission following behavioral (e.g., training) or therapeutic interventions, pathology or aging and therefore allows a better understanding of neural processes that underlie movement control and motor learning.

The aim of the present study was to assess changes in transmission at the corticomotoneuronal synapses in humans after repetitive transcranial magnetic stimulation. For this purpose, an electrophysiological method is introduced that allows assessment of pathway specific corticospinal transmission, i.e. differentiation of fast, direct corticospinal pathways from polysynaptic connections.

非侵入性评估人类皮质神经元传播的变化

The corticospinal pathway is the major pathway connecting the brain with the muscles and is therefore highly relevant for movement control and motor ...

Denver Papillae Protocol for Objective Analysis of Fungiform Papillae

The goal of the Denver Papillae Protocol is to use a dichotomous key to define and prioritize the characteristics of fungiform papillae (FP) to ensure consistent scoring between scorers. This protocol builds off of a need that has arisen from the last two decades of taste research using FP as a proxy for taste pore density. FP density has historically been analyzed using Miller &#38; Reedy&#8217;s 1990 characterizations of their morphology: round, stained lighter, large, and elevated. In this work, the authors forewarned that stricter definitions of FP morphology needed to be outlined. Despite this call to action, follow up literature has been scarce, with most studies continuing to cite Miller &#38; Reedy&#8217;s original work. Consequently, FP density reports have been highly variable and, combined with small sample sizes, may contribute to the discrepant conclusions on the role of FP in taste sensitivity. The Genetics of Taste Lab explored this apparent inconsistency in counting and found that scorers were individually prioritizing the importance of these characteristics differently and had no guidance for when a papilla had some, but not all, of the reported qualities of FP. The result of this subjectivity is highly variable FP counts of the same tongue image. The Denver Papillae Protocol has been developed to remedy this consequence through use of a dichotomous key that further defines and prioritizes the importance of the characteristics put forth by Miller &#38; Reedy. The proposed method could help create a standard way to quantify FP for researchers in the field of taste and nutritional studies.

Here we present a protocol to measure fungiform papilla density from digital photographs. This method builds prioritization and objective characteristic metrics into the original descriptive work on fungiform papillae by Miller &#38; Reedy (1990).

为菌状乳头的客观分析丹佛乳头协议

The goal of the Denver Papillae Protocol is to use a dichotomous key to define and prioritize the characteristics of fungiform papillae (FP) to ensure ...

A Rapid Food-Preference Assay in Drosophila

To select food with nutritional value while avoiding the consumption of harmful agents, animals need a sophisticated and robust taste system to evaluate their food environment. The fruit fly, Drosophila melanogaster, is a genetically tractable model organism that is widely used to decipher the molecular, cellular, and neural underpinnings of food preference. To analyze fly food preference, a robust feeding method is needed. Described here is a two-choice feeding assay, which is rigorous, cost-saving, and fast. The assay is Petri-dish-based and involves the addition of two different foods supplemented with blue or red dye to the two halves of the dish. Then, ~70 prestarved, 2-4-day-old flies are placed in the dish and&#160;allowed to choose between blue and red foods in the dark for about 90 min. Examination of the abdomen of each fly is followed by the calculation of the preference index. In contrast to multiwell plates, each Petri dish takes only ~20 s to fill and saves time and effort. This feeding assay can be employed to quickly determine whether flies like or dislike a particular food.

A Rapid Food-Preference Assay in Drosophila

We present a protocol for a two-choice feeding assay for flies. This feeding assay is fast and easy to run and is suitable not only for small-scale laboratory research, but also for high-throughput behavioral screens in flies.

果蝇的快速食物偏好分析

To select food with nutritional value while avoiding the consumption of harmful agents, animals need a sophisticated and robust taste system to evaluate ...