这个项目是由自然科学和工程研究理事会，加拿大（355881）和加拿大创新基金会的赠款支持。

*最重要的步骤。 1。设备和软件<ol><li>配备两个超轻型的，可伸缩的一侧或食物盛器（ 见表1）定位杆1鼠标操作性商会进行了训练和测试。位于上述各杠杆刺激光，并在笼子的顶部有一个房子光。颗粒机是位于外室。每个分庭内的换气扇声音衰减的房间内。钱伯斯是连接与MedPC四，软件到PC计算机上通过的"智能控制"界面内阁。 </li></ol> 注:当超轻型杠杆不可，建议鼻子戳用于小鼠研究的操作性响应。然而，有两个可供选择，我们建议选择更可靠的鼻子戳反应可以作为一个动机措施超轻杠杆é引发探索嗅探。 <ol start="2"><li>用于运行操作性任务MedPC方案都写在Medstate符号代码，用户可以了解编程手册提供的帮助。编程为用户提供灵活性，修改现有的代码，或产生独特的实验参数。某些代码可以得到免费从医学协会"Medstate符号库" <a href="http://www.mednr.com" target="_blank">http://www.mednr.com</a> 。从医学协会可以购买额外的代码。 </li></ol> 2。鼠标驯化和食品的限制<ol><li>抵达后小鼠组安置至少10天12小时的反向光/暗周期，以适应环境，并提供标准饲料和水的随意访问。在我们的例子中，我们用成年C57BL / 6小鼠从查尔斯河（圣常，质量控制）购买。所有的鼠标操作和行为实验是​​在黑暗的周期。所有亲涉及使用动物的cedures由CRCHUM动物保健委员会批准。 </li><li>小鼠随后被单独安置和食物限制（他们每天的食物摄入量的70％），直到他们达到90-95％，饲养体重，以便收购杠杆记者回应。小鼠在家里笼提供会话终止后，他们每天的食品配额。达到5-10％的体重损失后，每天的食物分配进行调整，以稳定为低体重训练期间的剩余。一旦小鼠已达到收购标准（见下文），他们随意喂养时间表。老鼠恢复失去体重约3-4天。 </li><li>小鼠在试验室进行处理的第一堂训练课之前连续3天。 10-15高脂肪和高糖（HFHS）颗粒进入家庭笼放置，以防止对食物恐新症对操作性performan潜在影响训练开始的前一天CE。我们使用20毫克HFHS，无尘，精密的食物颗粒（14毫克，也可）含48.9％千卡脂肪（生物SERV，Frenchtown，新泽西州）。 </li></ol> 3。操作性训练<ol><li>最初的训练小鼠按一个固定比率（FR）-1加固进度，即单杆按下插座引出的食物颗粒交货的杠杆。只有一杆被指定为"主动"（触发运送食物奖励）和左，右杠杆分配之间的小鼠抵消。重要的是，添加至少5秒超时（TO）FR1号的时间表（FR1/TO-5），在此期间，额外的杠杆按不会导致交货的食物颗粒。这个期限，使小鼠的时间消耗的食物颗粒，但是更长的时间超时如果实验者认为它是必要的小鼠模型，可以使用。每个FR培训会议持续1小时或50颗粒时已交付。 </li><li> *收购标准:&lt;/ STRONG&gt;用于确定收购粮食保持响应操作性的标准包括:（1）赚取的积极响应和奖励的最低数量;（2）（3）的活动和非活动的杠杆之间的歧视措施，并与会议性能稳定。小鼠表现出主动与非杠杆≥3:1的歧视，并取得了连续三届会议≥20％奖励被视为实现收购标准10。我们已经指出，C57BL6小鼠的大部分（75％）需要约7-10天的培训，以达到收购标准。小鼠，没有达到通过这次收购标准，接受进一步的训练额外的5-7天。我们排除测试（〜5％的情况下）小鼠时，有没有额外的第5个训练日的进度。重要的是要牢记操作性收购受损可能是某些药物和遗传干预的结果，因此，应记录。&lt;/ LI&gt; <li>继连续三年获得≥20颗粒会议，日程安排增加活跃的5杆印刷机引发的粮食颗粒交货FR5/TO-5秒。培训的FR5日程上持续了三天。 </li><li>小鼠，然后训练累进比率（PR）加固进度。理查德森和Roberts（1996）11％计算公关测试过程中的响应比时间表可以使用下列公式计算（四舍五入到最接近的整数）: = [5 E（R * 0.2）] - 5 其中 R等于已经获得食物奖励加1号（即下增强剂）。因此，需要获得食物奖励，按照订单的响应:1，2，4，6，9，12，15，20，25，32，40，50，62，77，95等。最终完成的比例是断点。 </li><li>公关会议最多持续1小时。未能在任何10分钟终止在会议期间按杠杆。 perfor钢筋公关时间表曼斯被认为是稳定时，奖励的数量获得了至少连续3天≤10％，在1个小时的会议偏离。 </li></ol> 4。进的比测试和验证<ol><li>为了验证食品保持操作性响应公关任务，可以评估断点响应以下操作，以调节食物奖励。一个相对简单的测试，开展粮食匮乏，这将增加食品鼠标的动机状态，从而增加断点。老鼠必须先进行测试期间随意喂养期间，直到实现稳定，性能基线的公关任务。最后的"基线"一天的食物后，一天从家里笼和老鼠，然后在24小时后开始快速测试的公关任务。 </li><li>我们还调查了抑制食欲的荷尔蒙leptin的外设管理是否会有相反的效果上断点阈值。瘦素（AF Parlow，国家激素和肽计划，NIDDK）溶解在无菌PBS。公关测试进行的PBS在一天的IP注射，然后注射瘦素（5毫克/公斤）的翌日。瘦素注射前一小时公关测试。断点比较采用配对t检验（GraphPad棱镜）。 </li></ol> 5。代表结果测试时，粮食匮乏的能力，以调节断点上一个加固的公关日程响应，我们期望显着增加，如在图1所示的断点。 24小时的食物匮乏的时期，在这个实验中产生的中断点〜3.6倍的增长相比，那些在自由摄食基线从而表明缺粮增加食物的奖赏效应的状态得到。相比之下，外设管理，厌食激素瘦素（5毫克/公斤，IP）前一小时到Testing应减少断点响应如图2所示。 <img alt="图1" src="/files/ftp_upload/3754/3754fig1.jpg" /> 图1。空腹增加食物的奖赏效应。 24 h快速显着增加了一个渐进的比率（PR）的增强小鼠的时间表（N = 4）断点。平均值±标准差; * P &lt;.05。 <img alt="图2" src="/files/ftp_upload/3754/3754fig2.jpg" /> 图2。瘦素的小鼠减少食物奖励。在小鼠（n = 4时），外周血瘦素管理（5毫克/公斤）下降公关断点响应阈值。平均值±标准差; * P &lt;0.05。

<ol>
	<li>Fulton, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Appetite+and+reward.">Appetite and reward.</a> Front. Neuroendocrinol. 31, 85-103 (2010).</li><li>Zheng, H., Lenard, N. R., Shin, A. C., Berthoud, H. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Appetite+control+and+energy+balance+regulation+in+the+modern+world:+reward-driven+brain+overrides+repletion+signals.">Appetite control and energy balance regulation in the modern world: reward-driven brain overrides repletion signals.</a> International journal of Obesity. 33, S8-S13 (2009).</li><li>Skinner, B. F. <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 Behavior of Organisms: An Experimental Analysis. , (1938).</li><li>Hodos, W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Progressive+ratio+as+a+measure+of+reward+strength.">Progressive ratio as a measure of reward strength.</a> Science. 134, 943-944 (1961).</li><li>Hodos, W., Kalman, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effects+of+increment+size+and+reinforcer+volume+on+progressive+ratio+performance.">Effects of increment size and reinforcer volume on progressive ratio performance.</a> J. Exp. Anal. Behav. 6, 387-392 (1963).</li><li>Stafford, D., LeSage, M. G., Glowa, J. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Progressive-ratio+schedules+of+drug+delivery+in+the+analysis+of+drug+self-administration:+a+review.">Progressive-ratio schedules of drug delivery in the analysis of drug self-administration: a review.</a> Psychopharmacology. , 139-169 (1998).</li><li>Glass, M. J., O'Hare, E., Cleary, J. P., Billington, C. J., Levine, A. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+effect+of+naloxone+on+food-motivated+behavior+in+the+obese+Zucker+rat.">The effect of naloxone on food-motivated behavior in the obese Zucker rat.</a> Psychopharmacology (Berl). 141, 378-384 (1999).</li><li>Brennan, K., Roberts, D. C., Anisman, H., Merali, Z. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Individual+differences+in+sucrose+consumption+in+the+rat:+motivational+and+neurochemical+correlates+of+hedonia.">Individual differences in sucrose consumption in the rat: motivational and neurochemical correlates of hedonia.</a> Psychopharmacology (Berl). 157, 269-276 (2001).</li><li>Vaughan, C., Moore, M., Haskell-Luevano, C., Rowland, N. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Food+motivated+behavior+of+melanocortin-4+receptor+knockout+mice+under+a+progressive+ratio+schedule.">Food motivated behavior of melanocortin-4 receptor knockout mice under a progressive ratio schedule.</a> Peptides. 27, 2829-2835 (2006).</li><li>Haluk, D. M., Wickman, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Evaluation+of+study+design+variables+and+their+impact+on+food-maintained+operant+responding+in+mice.">Evaluation of study design variables and their impact on food-maintained operant responding in mice.</a> Behav. Brain Res. 207, 394-401 (2010).</li><li>Richardson, N. R., Roberts, D. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Progressive+ratio+schedules+in+drug+self-administration+studies+in+rats:+a+method+to+evaluate+reinforcing+efficacy.">Progressive ratio schedules in drug self-administration studies in rats: a method to evaluate reinforcing efficacy.</a> J. Neurosci. Methods. 66, 1-11 (1996).</li><li>Jewett, D. C., Cleary, J., Levine, A. S., Schaal, D. W., Thompson, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effects+of+neuropeptide+Y,+insulin,+2-deoxyglucose,+and+food+deprivation+on+food-motivated+behavior.">Effects of neuropeptide Y, insulin, 2-deoxyglucose, and food deprivation on food-motivated behavior.</a> Psychopharmacology (Berl). 120, 267-271 (1995).</li><li>Fulton, S., Woodside, B., Shizgal, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Modulation+of+brain+reward+circuitry+by+leptin.">Modulation of brain reward circuitry by leptin.</a> Science. 287, 125-128 (2000).</li><li>Figlewicz, D. P., Bennett, J. L., Naleid, A. M., Davis, C., Grimm, J. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Intraventricular+insulin+and+leptin+decrease+sucrose+self-administration+in+rats.">Intraventricular insulin and leptin decrease sucrose self-administration in rats.</a> Physiol. Behav. 89, 611-616 (2006).</li></ol>

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操作性条件反射的任务提供了一种有效的手段，以评估食物的动机性质的变化。本文许可证训练小鼠食物奖励杠杆记者和稳定详述的实验程序响应的加固公关时间表。 测量消耗的食物量不提供相同的读数，因为并不是所有的因素，影响食物的摄入，调节食物的奖赏效应，努力为基础的应对措施。例如，2 -脱氧葡萄糖对大鼠外周血管理消耗利用的葡萄糖增加量免费提供食用的食品，但未能改变破发点，为12食品的公关任务。公关断点响应，也可以反映在食物奖励的质量变化。例如，破发点的正相关，与蔗糖浓度展示方式如下甜味8，增加蔗糖的动机</s达&gt;。在这里，我们表明，24小时剥夺食物大大增加公关小鼠HFHS食品断点。相比之下，我们发现抑制食欲的荷尔蒙leptin的外设管理，显着地降低小鼠为HFHS食品断点。这些研究结果与以前报道的行动粮食匮乏和12-14大鼠瘦素奖励阈值一致。 使用此协议时，重要的是要牢记，粮食收购保持定期培训（每1-2天），并通过实施食品的限制方案，通过增加鼠标的动机状态明显增强操作性响应。此外，确保小鼠表现出相对稳定的一个加固的时间表响应之前移动到更复杂的时间表（如FR1号到FR5），以减少在测试过程中的可变性能的关键。尽管这些措施的应用，这是不寻常，以获得一些变异特别是在不同小鼠之间。出于这个原因，我们强烈建议使用内（重复测量）设计时，可以删除跨学科变异。虽然内受测试调查稳定的遗传改变（即淘汰赛与对照组）的可能，它可以用来检查药物，病变或急性遗传操作（例如，optogenetic）的影响。 肥胖与肥胖有关的疾病是一个日益严重的问题，增加了紧迫感，以确定涉及的分子和神经通路。越来越多的证据突出的重要作用，在食物渴求的大脑奖励电路，增加热量的摄入和体重增加。公关任务是一个有价值的工具，用于测量食物奖赏效应的影响，神经生物学，遗传或药理操纵。

<html><table border="1"><tbody><tr><td> 项目 </td><td> 目录编号 </td><td> 公司 </td><td> 评论 </td></tr><tr><td>小鼠操作性商会</td><td> env-009A的</td><td> <a href="http://www.med-associates.com/" target="_blank">MED公司，公司</a> </td><td> MED联营公司 PO箱319 圣奥尔本斯，佛蒙特州05478， 美国 电话:（802）527-2343 </td></tr><tr><td> 8 inputs/16输入智能 控制连接面板</td><td> SG-716B </td><td> <a href="http://www.med-associates.com/" target="_blank">MED公司，公司</a> </td><td></td></tr><tr><td>超轻型伸缩 鼠标杆</td><td> env-312的</td><td> <a href="http://www.med-associates.com/" target="_blank">MED公司，公司</a> </td><td></td></tr><tr><td>佩莱饮水机</td><td> ENV-203 </td><td>MED-associates.com /"目标="_blank"&gt; MED Associates公司</td><td></td></tr><tr><td>楼光</td><td> env-215的</td><td> <a href="http://www.med-associates.com/" target="_blank">MED公司，公司</a> </td><td></td></tr><tr><td>刺激光</td><td> ENV-221 </td><td> <a href="http://www.med-associates.com/" target="_blank">MED公司，公司</a> </td><td></td></tr><tr><td>无尘精密20 毫克高脂肪及高糖 颗粒</td><td>架F-06649 </td><td>生物-​​Serv的</td><td>生物-​​Serv的 一街8号，1套房 frenchtown，NJ 08825，USA 电话:（800）-996-9908 </td></tr></tbody></table> 表1。具体的设备和材料。

progressive-ratio responding for palatable high-fat and high-sugar food in mice

含有丰富的脂肪和糖的食物，大大有助于过度饮食和肥胖率的不断上升。可口的食物消费，可以产生有益的作用，加强行动的成果协会和加强未来的行为，在取得这些食物。越来越多的证据，能量密度高的食物奖赏效应发挥暴饮暴食和肥胖的发展产生深远的作用，有研究的基因，分子和神经电路，调节食物奖励1,2的兴趣高涨。可以测量的意愿工作，以取得他们如在操作性条件反射的任务3，研究不同的刺激有益的影响。收购和食物奖励措施的操作性模型，在获取食物的自愿行为反应。奖励实力的一个常用的措施是在公关ţ4,5操作性的过程称为累进比率（PR）的加固进度。问，这个问题需要作出操作性反应的越来越多，为每个连续的奖励。的Hodos（1961）的开创性研究表明，作出反应，获得最后的奖励数量，被称为断点，作为奖励强度指数4。尽管操作性的程序，仅在响应速度的变动不能分开奖励强度的变化，表现能力的改变，来自公关时间表的断点是食物的奖赏效应的验证措施。公关任务已被广泛用于评估药物滥用和老鼠的食物（如6-8）有益的影响，但程度较轻，在小鼠9。遗传工程小鼠和饮食诱导的肥胖小鼠模型使用的增加加剧了在老鼠的食物奖励的行为措施的需求。在本文中，我们详细的材料和程序，用来训练老鼠高脂肪回应（杠杆新闻）和高糖食物颗粒上加固公关时间表。我们表明，断点的响应阈值增加剥夺后急性食物，并减少与抑制食欲的荷尔蒙leptin的外设管理，从而验证了使用这种小鼠的食品操作性范式。

Watch this Scientific Journal Video about Progressive-ratio Responding for Palatable High-fat and High-sugar Food in Mice at JoVE.com

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

本报告详细说明来衡量高脂肪食物的小鼠的奖赏效应的协议，采用累进比例操作性条件反射的任务。

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 ...

progressive-ratio-responding-for-palatable-high-fat-high-sugar-food

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22.0K 次观看.  University of Montreal. 本报告详细说明来衡量高脂肪食物的小鼠的奖赏效应的协议，采用累进比例操作性条件反射的任务。

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

T-maze Forced Alternation and Left-right Discrimination Tasks for Assessing Working and Reference Memory in Mice

Forced alternation and left-right discrimination tasks using the T-maze have been widely used to assess working and reference memory, respectively, in rodents. In our laboratory, we evaluated the two types of memory in more than 30 strains of genetically engineered mice using the automated version of this apparatus. Here, we present the modified T-maze apparatus operated by a computer with a video-tracking system and our protocols in a movie format. The T-maze apparatus consists of runways partitioned off by sliding doors that can automatically open downward, each with a start box, a T-shaped alley, two boxes with automatic pellet dispensers at one side of the box, and two L-shaped alleys. Each L-shaped alley is connected to the start box so that mice can return to the start box, which excludes the effects of experimenter handling on mouse behavior. This apparatus also has an advantage that in vivo microdialysis, in vivo electrophysiology, and optogenetics techniques can be performed during T-maze performance because the doors are designed to go down into the floor. In this movie article, we describe T-maze tasks using the automated apparatus and the T-maze performance of &alpha;-CaMKII+/- mice, which are reported to show working memory deficits in the eight-arm radial maze task. Our data indicated that &alpha;-CaMKII+/- mice showed a working memory deficit, but no impairment of reference memory, and are consistent with previous findings using the eight-arm radial maze task, which supports the validity of our protocol. In addition, our data indicate that mutants tended to exhibit reversal learning deficits, suggesting that &alpha;-CaMKII deficiency causes reduced behavioral flexibility. Thus, the T-maze test using the modified automatic apparatus is useful for assessing working and reference memory and behavioral flexibility in mice.

This article presents the protocol of T-maze tests using a modified automated apparatus for assessing the learning and memory functions in mice.

T-迷宫强迫交替和评估工作，并参考在小鼠记忆的左，右的歧视任务

Forced alternation and left-right discrimination tasks using the T-maze have been widely used to assess working and reference memory, respectively, in ...

科研

神经科学

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

Emerging evidence from a number of laboratories indicates that humans have the ability to identify fatty acids in the oral cavity, presumably via fatty acid receptors housed on taste cells. Previous research has shown that an individual&#39;s oral sensitivity to fatty acid, specifically oleic acid (C18:1) is associated with body mass index (BMI), dietary fat consumption, and the ability to identify fat in foods. We have developed a reliable and reproducible method to assess oral chemoreception of fatty acids, using a milk and C18:1 emulsion, together with an ascending forced choice triangle procedure. In parallel, a food matrix has been developed to assess an individual&#39;s ability to perceive fat, in addition to a simple method to assess fatty food liking. As an added measure tongue photography is used to assess papillae density, with higher density often being associated with increased taste sensitivity.

Oral chemoreception of fatty acids and the association with diet and fatty food preferences may enable the identification of mechanisms involved with the development of obesity and why dietary changes may be difficult for many individuals.

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 ...

A Simple and Inexpensive Method for Determining Cold Sensitivity and Adaptation in Mice

Cold hypersensitivity is a serious clinical problem, affecting a broad subset of patients and causing significant decreases in quality of life. The cold plantar assay allows the objective and inexpensive assessment of cold sensitivity in mice, and can quantify both analgesia and hypersensitivity. Mice are acclimated on a glass plate, and a compressed dry ice pellet is held against the glass surface underneath the hindpaw. The latency to withdrawal from the cooling glass is used as a measure of cold sensitivity.
Cold sensation is also important for survival in regions with seasonal temperature shifts, and in order to maintain sensitivity animals must be able to adjust their thermal response thresholds to match the ambient temperature. The Cold Plantar Assay (CPA) also allows the study of adaptation to changes in ambient temperature by testing the cold sensitivity of mice at temperatures ranging from 30 &#176;C to 5 &#176;C. Mice are acclimated as described above, but the glass plate is cooled to the desired starting temperature using aluminum boxes (or aluminum foil packets) filled with hot water, wet ice, or dry ice. The temperature of the plate is measured at the center using a filament T-type thermocouple probe. Once the plate has reached the desired starting temperature, the animals are tested as described above.
This assay allows testing of mice at temperatures ranging from innocuous to noxious. The CPA yields unambiguous and consistent behavioral responses in uninjured mice and can be used to quantify both hypersensitivity and analgesia. This protocol describes how to use the CPA to measure cold hypersensitivity, analgesia, and adaptation in mice.

The Cold Plantar Assay (CPA) measures cold responsiveness between 30 &#176;C and 5 &#176;C, and can also measure cold adaptation. This protocol describes how to use the CPA to measure cold hypersensitivity, analgesia, and adaptation in mice.

测定冷灵敏度和适应小鼠一种简单廉价的方法

Cold hypersensitivity is a serious clinical problem, affecting a broad subset of patients and causing significant decreases in quality of life.  The cold ...

The CApillary FEeder Assay Measures Food Intake in Drosophila melanogaster

For most animals, feeding is an essential behavior for securing survival, and it influences development, locomotion, health and reproduction. Ingestion of the right type and quantity of food therefore has a major influence on quality of life. Research on feeding behavior focuses on the underlying processes that ensure actual feeding and unravels the role of factors regulating internal energy homeostasis and the neuronal bases of decision-making. The model organism Drosophila melanogaster, with its great variety of genetically traceable tools for labeling and manipulating single neurons, allows mapping of neuronal networks and identification of molecular signaling cascades involved in the regulation of food intake. This report demonstrates the CApillary FEeder assay (CAFE) and shows how to measure food intake in a group of flies for time spans ranging from hours to days. This easy-to-use assay consists of glass capillaries filled with liquid food that flies can freely access and feed on. Food consumption in the assay is accurately determined using simple measurement tools. Herein we describe step-by-step the method from setup to successful execution of the CAFE assay, and provide practical examples to analyze the food intake of a group of flies under controlled conditions. The reader is guided through possible limitations of the assay, and advantages and disadvantages of the method compared to other feeding assays in D. melanogaster are evaluated.

The CApillary FEeder Assay Measures Food Intake in Drosophila melanogaster

The CApillary FEeder (CAFE) assay is a simple, budget-friendly, highly reliable method for investigating mechanisms underlying food intake. Used with the highly versatile genetic model organism Drosophila melanogaster, it provides a powerful means of gaining new insights into regulatory mechanisms of food intake.

毛细管馈线分析测量的摄食<em&gt;果蝇</em

For most animals, feeding is an essential behavior for securing survival, and it influences development, locomotion, health and reproduction. Ingestion of ...

Assessment of Dopaminergic Homeostasis in Mice by Use of High-performance Liquid Chromatography Analysis and Synaptosomal Dopamine Uptake

Dopamine (DA) is a modulatory neurotransmitter controlling motor activity, reward processes and cognitive function. Impairment of dopaminergic (DAergic) neurotransmission is strongly associated with several central nervous system-associated diseases such as Parkinson&#39;s disease, attention-deficit-hyperactivity disorder and drug addiction1,2,3,4. Delineating disease mechanisms involving DA imbalance is critically dependent on animal models to mimic aspects of the diseases, and thus protocols that assess specific parts of the DA homeostasis are important to provide novel insights and possible therapeutic targets for these diseases.
Here, we present two useful experimental protocols that when combined provide a functional read-out of the DAergic system in mice. Biochemical and functional parameters on DA homeostasis are obtained through assessment of DA levels and dopamine transporter (DAT) functionality5. When investigating the DA system, the ability to reliably measure endogenous levels of DA from adult brain is essential. Therefore, we present how to perform high-performance liquid chromatography (HPLC) on brain tissue from mice to determine levels of DA. We perform the experiment on tissue from dorsal striatum (dStr) and nucleus accumbens (NAc), but the method is also suitable for other DA-innervated brain areas.
DAT is essential for reuptake of DA into the presynaptic terminal, thereby controlling the temporal and spatial activity of released DA. Knowing the levels and functionality of DAT in the striatum is of major importance when assessing DA homeostasis. Here, we provide a protocol that allows to simultaneously deduce information on surface levels and function using a synaptosomal6 DA uptake assay.
Current methods combined with standard immunoblotting protocols provide the researcher with relevant tools to characterize the DAergic system.

Synaptosomal dopamine uptake and high-performance liquid chromatography analysis represent experimental tools to investigate dopamine homeostasis in mice by assessing the function of the dopamine transporter and levels of dopamine in striatal tissue, respectively. Here we present protocols to measure dopamine tissue content and assess the functionality of the dopamine transporter.

高效液相色谱分析和触多巴胺摄取对小鼠多巴胺能稳态的评价

Dopamine (DA) is a modulatory neurotransmitter controlling motor activity, reward processes and cognitive function. Impairment of dopaminergic (DAergic) ...

An In Ovo Model for Testing Insulin-mimetic Compounds

Elevated blood glucose levels in type 2 diabetes mellitus (T2DM), a complex and multifactorial metabolic disease, are caused by insulin resistance and &#946;-cell failure. Various strategies, including the injection of insulin or the usage of insulin-sensitizing drugs, were pursued to treat T2DM or at least reduce the symptoms. In addition, the application of herbal compounds has attracted increasing attention. Thus, it is necessary to find efficient test systems to identify and characterize insulin-mimetic compounds. Here we developed a modified chick embryo model, which enables testing of synthetic compounds and herbal extracts with insulin-mimetic properties. Using a fluorescence microscopy-based primary screen, which quantifies the translocation of Glucose transporter 4 (Glut4) to the plasma membrane, we were able to identify compounds, mainly herbal extracts, which lead to an increase of intracellular glucose concentrations in adipocytes. However, the efficacy of these substances requires further verification in a living organism. Thus, we used an in-ovo approach to identify their blood glucose-reducing properties. The approval by an ethics committee is not needed since the use of chicken embryos during the first two-thirds of embryonic development is not considered an animal experiment. Here, the application of this model is described in detail.

An In Ovo Model for Testing Insulin-mimetic Compounds

In this study, we describe an in-ovo system as a promising tool to test insulin-mimetic compounds in a living organism. The main advantages include adequate throughput rates and acceptable costs, which enable the identification of phytochemicals with insulin-mimetic properties.

用于测试胰岛素模拟化合物的蛋模型

Elevated blood glucose levels in type 2 diabetes mellitus (T2DM), a complex and multifactorial metabolic disease, are caused by insulin resistance and ...

Optogenetic Entrainment of Hippocampal Theta Oscillations in Behaving Mice

Extensive data on relationships of neural network oscillations to behavior and organization of neuronal discharge across brain regions call for new tools to selectively manipulate brain rhythms. Here we describe an approach combining projection-specific optogenetics with extracellular electrophysiology for high-fidelity control of hippocampal theta oscillations (5-10 Hz) in behaving mice. The specificity of the optogenetic entrainment is achieved by targeting channelrhodopsin-2 (ChR2) to the GABAergic population of medial septal cells, crucially involved in the generation of hippocampal theta oscillations, and a local synchronized activation of a subset of inhibitory septal afferents in the hippocampus. The efficacy of the optogenetic rhythm control is verified by a simultaneous monitoring of the local field potential (LFP) across lamina of the CA1 area and/or of neuronal discharge. Using this readily implementable preparation we show efficacy of various optogenetic stimulation protocols for induction of theta oscillations and for the manipulation of their frequency and regularity. Finally, a combination of the theta rhythm control with projection-specific inhibition addresses the readout of particular aspects of the hippocampal synchronization by efferent regions.

We describe the use of optogenetics and electrophysiological recordings for selective manipulations of hippocampal theta oscillations (5-10 Hz) in behaving mice. The efficacy of the rhythm entrainment is monitored using local field potential. A combination of opto- and pharmacogenetic inhibition addresses the efferent readout of hippocampal synchronization.

Optogenetic 在行为小鼠海马θ振荡中的作用

Extensive data on relationships of neural network oscillations to behavior and organization of neuronal discharge across brain regions call for new tools ...

A Protocol for Transcranial Photobiomodulation Therapy in Mice

Transcranial photobiomodulation is a potential innovative noninvasive therapeutic approach for improving brain bioenergetics, brain function in a wide range of neurological and psychiatric disorders, and memory enhancement in age-related cognitive decline and neurodegenerative diseases. We describe a laboratory protocol for transcranial photobiomodulation therapy (PBMT) in mice. Aged BALB/c mice (18 months old) are treated with a 660 nm laser transcranially, once daily for 2 weeks. Laser transmittance data shows that approximately 1% of the incident red light on the scalp reaches a 1 mm depth from the cortical surface, penetrating the dorsal hippocampus. Treatment outcomes are assessed by two methods: a Barnes maze test, which is a hippocampus-dependent spatial learning and memory task evaluation, and measuring hippocampal ATP levels, which is used as a bioenergetics index. The results from the Barnes task show an enhancement of the spatial memory in laser-treated aged mice when compared with age-matched controls. Biochemical analysis after laser treatment indicates increased hippocampal ATP levels. We postulate that the enhancement of memory performance is potentially due to an improvement in hippocampal energy metabolism induced by the red laser treatment. The observations in mice could be extended to other animal models since this protocol could potentially be adapted to other species frequently used in translational neuroscience, such as rabbit, cat, dog, or monkey. Transcranial photobiomodulation is a safe and cost-effective modality which may be a promising therapeutic approach in age-related cognitive impairment.

Photobiomodulation therapy is an innovative noninvasive modality for the treatment of a wide range of neurological and psychiatric disorders and can also improve healthy brain function. This protocol includes a step-by-step guide to performing brain photobiomodulation in mice by transcranial light delivery, which can be adapted for use in other laboratory rodents.

小鼠经颅光生物调节治疗方案

Transcranial photobiomodulation is a potential innovative noninvasive therapeutic approach for improving brain bioenergetics, brain function in a wide ...

Data Acquisition and Analysis In Brainstem Evoked Response Audiometry In Mice

Brainstem evoked response audiometry (BERA) is of central relevance in the clinical neurophysiology. As other evoked potential (EP) techniques, such as visually evoked potentials (VEPs) or somatosensory evoked potentials (SEPs), the auditory evoked potentials (AEPs) are triggered by the repetitive presentation of identical stimuli, the electroencephalographic (EEG) response of which is subsequently averaged resulting in distinct positive (p) and negative (n) deflections. In humans, both the amplitude and the latency of individual peaks can be used to characterize alterations in synchronization and conduction velocity in the underlying neuronal circuitries. Importantly, AEPs are also applied in basic and preclinical science to identify and characterize the auditory function in pharmacological and genetic animal models. Even more, animal models in combination with pharmacological testing are utilized to investigate for potential benefits in the treatment of sensorineural hearing loss (e.g., age- or noise-induced hearing deficits). Here we provide a detailed and integrative description of how to record auditory brainstem-evoked responses (ABRs) in mice using click and tone-burst application. A specific focus of this protocol is on pre-experimental animal housing, anesthesia, ABR recording, ABR filtering processes, automated wavelet-based amplitude growth function analysis, and latency detection.

Brainstem evoked response audiometry is an important tool in clinical neurophysiology. Nowadays, brainstem evoked response audiometry is also applied in the basic science and preclinical studies involving both pharmacological and genetic animal models. Here we provide a detailed description of how auditory brainstem responses can be successfully recorded and analyzed in mice.

脑干中的数据采集与分析 在小鼠中引发响应音频测量

Brainstem evoked response audiometry (BERA) is of central relevance in the clinical neurophysiology. As other evoked potential (EP) techniques, such as ...

Quantitative Measurement of Intrathecally Synthesized Proteins in Mice

Cerebrospinal fluid (CSF), a fluid found in the brain and the spinal cord, is of great importance to both basic and clinical science. The analysis of the CSF protein composition delivers crucial information in basic neuroscience research as well as neurological diseases. One caveat is that proteins measured in CSF may derive from both intrathecal synthesis and transudation from serum, and protein analysis of CSF can only determine the sum of these two components. To discriminate between protein transudation from the blood and intrathecally produced proteins in animal models as well as in humans, CSF protein profiling measurements using conventional protein analysis tools must include the calculation of the albumin CSF/serum quotient (Qalbumin), a marker of the integrity of the blood-brain interface (BBI), and the protein index (Qprotein/Qalbumin), an estimate of intrathecal protein synthesis. This protocol illustrates the entire procedure, from CSF and blood collection to quotients and indices calculations, for the quantitative measurement of intrathecal protein synthesis and BBI impairment in mouse models of neurological disorders.

Elevated spinal fluid protein levels can either be the result of diffusion of plasma protein across an altered blood-brain barrier or intrathecal synthesis. An optimized testing protocol is presented in this article that helps to discriminate both cases and provides quantitative measurements of intrathecally synthesized proteins.

小鼠体内合成蛋白的定量测量

Cerebrospinal fluid (CSF), a fluid found in the brain and the spinal cord, is of great importance to both basic and clinical science. The analysis of the ...

累进比率响应小鼠可口的高脂肪和高糖食品

摘要

探索更多视频

此视频中的章节

T-迷宫强迫交替和评估工作，并参考在小鼠记忆的左，右的歧视任务

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

测定冷灵敏度和适应小鼠一种简单廉价的方法

毛细管馈线分析测量的摄食

高效液相色谱分析和触多巴胺摄取对小鼠多巴胺能稳态的评价

用于测试胰岛素模拟化合物的蛋模型

Optogenetic 在行为小鼠海马θ振荡中的作用

小鼠经颅光生物调节治疗方案

脑干中的数据采集与分析在小鼠中引发响应音频测量

小鼠体内合成蛋白的定量测量

累进比率响应小鼠可口的高脂肪和高糖食品

摘要

探索更多视频

此视频中的章节

T-迷宫强迫交替和评估工作，并参考在小鼠记忆的左，右的歧视任务

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

测定冷灵敏度和适应小鼠一种简单廉价的方法

毛细管馈线分析测量的摄食

高效液相色谱分析和触多巴胺摄取对小鼠多巴胺能稳态的评价

用于测试胰岛素模拟化合物的蛋模型

Optogenetic 在行为小鼠海马θ振荡中的作用

小鼠经颅光生物调节治疗方案

脑干中的数据采集与分析 在小鼠中引发响应音频测量

小鼠体内合成蛋白的定量测量

脑干中的数据采集与分析在小鼠中引发响应音频测量