这项工作是由国家卫生部批准R01 GM54408授予立法会格里菲斯学院的支持。我们要感谢他对所有帮助我们的方腔设计和设置我们的实验中，弗兰克梅洛在布兰代斯大学的机加工车间建设我们的商会，丹瓦伦特和蒂姆Lebestky关于有助于分析问题的谈话弗雷德沃尔夫。

第1部分：苍蝇的饲养和管理<ol><li>苍蝇应该是生长在含有酵母的标准媒体瓶。您将需要大量的苍蝇，所以需要设立相应的十字架。苍蝇应在25℃生长在一个12小时的光：暗周期</li><li>苍蝇应收集羽化后不久（1-3天）。苍蝇可与使用二氧化碳扩散的阶段，并应包含蒸压或酵母媒体的测试管排序。我们使用雄性10只，存储到试管中。 </li><li>允许使用二氧化碳后运行行为实验前至少一天或两天。我们运行我们的行为实验中的苍蝇以下羽化3-5天。 </li><li>检测应该总是做相同的2-3小时的时间窗口每一天内，以避免昼夜节律问题。 </li></ol>第2部分：设置在受控环境中的跟踪系统<ol><li>所有的实验都保持一个恒定温度25 º C，湿度70％的环境中，房间。 </li><li>暂停对该地区的数码摄像机要记录（摄像机朝下）。由于我们的跟踪软件为基础的对比，我们会暂停灯箱相机。相机记录影片直接到戴尔电脑通过FireWire连接。我们使用了夏普的数字视频摄像机，视频采集挂接到一个使用Windows MovieMaker的戴尔计算机（Vista版本）。 </li><li>空气脉冲将在这个协议管理，使空气流动的来源，必须在房间里。使用橡胶管连接气源为空气过滤器活性炭过滤器。 </li><li>使用更多的橡胶管，连接另一端的碳过滤器，通过一个中空的橡胶塞锥形瓶中，并填写烧瓶中，约半英寸的水。这将湿润的空气。 </li><li>烧瓶，也应该有一个手枪，这将连接到一个Y形阀，通过橡皮管。 </li><li> Y型阀的一个分支应该被连接到一个流量计。其他部门应提供空气平方米的运动室。 </li></ol>第3部分：运动室提供苍蝇<ol><li>平方米的运动室Wolf 等人设计的基础上从2002 年 1，小降盘此外，商会内的飞行，以防止苍蝇。商会应采取外，应彻底消灭所有的组件，用70％乙醇和使用前干燥。 </li><li>麻醉前，立即进行行为检测可能妥协的性能。为了避免这种情况，苍蝇轻轻敲成一个小插座使用一个漏斗分。为了让大小合适的插座，附加一个蓝色的枪头（P1000）的一个漏斗年底。用剪刀切断枪头的​​，开放的大飞通过足够。 </li><li>我们的房间的顶部是一个小块有机玻璃边缘周围的螺丝固定。被删除，当螺丝孔，可用于提供苍蝇进入会议厅。有机玻璃顶部的螺丝孔，以便直接在位于内宅，而不是一个螺丝钉的位置一般位于的位置。试管中含有的苍蝇，放在倒置的漏斗，然后应放置到螺丝孔。 </li><li>不要移动漏斗，因为这可能伤害或损坏的苍蝇。相反，轻轻的一声整个腔，漏斗和所有，直到所有的苍蝇箱内。这敲应该做的鼠标垫，吸收这一撞冲击。当苍蝇在里面，取出漏斗，重新定位的螺丝床有机玻璃顶部。然后重新拧入地方有机玻璃顶部。 </li></ol>第4部分：跑步运动检测<ol><li>一旦苍蝇正确加载，让他们适应环境在30分钟的会议厅举行会议。这驯化期间，应采取在控制的环境中（25 ° C，湿度70％），和室应放在灯箱上（应打开）。关闭其他房间的灯熄灭。 </li><li>驯化期间，Y型阀开关在空气流通系统，如空气流量计只会流。打开空气，坡道空气所需的速度（4.0-6.0升/分钟）。我们一般使用5.0-5.5升/分钟，但在这个范围内的任何速度会的工作。 </li><li>一旦达到所需的空速，Y型阀切换，使空气流动的运动室。时间为15秒的空气脉冲，然后突然打开空气。当您打开空气开关的相机记录模式。 （这部分需要练习做一次）。 </li><li>所需的时间后停止录音并保存。我们记录每秒10帧（至少8个基因型试验），30秒的试验。 </li></ol>第5部分：视频分析<ol><li>我们使用动态图像分析系统（DIAS）3.2软件议案追踪2。为了使用这种软件，我们首先我们的视频转换成AVI文件格式的QuickTime 7.5.5。 </li><li>要跟踪对比的基础苍蝇，我们使用“阈值自动跟踪”功能。然后，我们使用“设为路径从跟踪”功能的数字化这些痕迹。 </li><li>每个飞输出的瞬时速度，我们使用“计算参数”的功能，产生一个数据库文件（DIAS特定）。使用此功能，我们也能顺利使用“5,15,60,15,5”杜克平滑窗口的数据。 </li><li>一旦这个信息是作为一个数据库文件的输出，它可以打开在Microsoft Excel中。我们使用MATLAB脚本来编译Excel内的瞬时速度，并计算为了解运动动态和布特结构所需的额外参数。 </li></ol>代表性的成果： 图1显示了从野生型广小号生物体（图1，左侧面板）的代表痕迹和苍蝇 dCASK基因，其中跨越了两个大的重叠缺失（DF（3R）X307和DF（3R）x313）（图1生成的空右图​​）。dCASK空苍蝇先前已被证明有使用布里丹的范式3的运动问题，并在我们的范例，广小号相比，他们表现出大大减少运动。我们始终运行野生型生物，以确保正常范围内的某一天，条件和行为。我们已经看到在不到5天的测试，偏离标准的反应。这些差异通常可以归结为我们的受控环境中的参数问题。 <img src="/files/ftp_upload/1096/figure_1.png" alt="" width="600" height="298" /> 图1。DIAS的产生既广小号野生型果蝇（上，左）和运动能力缺陷dCASK空苍蝇重叠缺失（上，右）所产生的痕迹。什么是录制的影片中看到的痕迹代表。图文并茂contrain 8-10苍蝇的痕迹在视频跟踪检测，30秒以下的空气脉冲运行。 <img src="/files/ftp_upload/1096/figure_2.png" alt="" width="600" height="438" /> 图2野生型果蝇的运动检测（以上），下面的空气脉冲（紫酒吧）和没有空气脉冲（蓝色）运行。在分析程序中计算所有参数，有两个条件之间没有显着差异（双尾学生t检验确定）。这表明，空气脉冲，苍蝇locomote通常在我们的设置。

<ol>
	<li>Wolf, F. W., Rodan, A. R., Tsai, L. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=High-Resolution+Analysis+of+Ethanol-Induced+Locomotor+Stimulation+in+Drosophila.">High-Resolution Analysis of Ethanol-Induced Locomotor Stimulation in Drosophila.</a> J Neurosci. 22 (24), 11035-11044 (2002).</li><li>Soll, D. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+use+of+computers+in+understanding+how+animal+cells+crawl.">The use of computers in understanding how animal cells crawl.</a> International review of cytology. 163, 43-104 (1995).</li><li>Martin, J. R., Ollo, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+new+Drosophila+Ca2+/calmodulin-dependent+protein+kinase+(Caki)+is+localized+in+the+central+nervous+system+and+implicated+in+walking+speed.">A new Drosophila Ca2+/calmodulin-dependent protein kinase (Caki) is localized in the central nervous system and implicated in walking speed.</a> The EMBO journal. 15 (8), 1865-1876 (1996).</li><li>Levine, J. D., Funes, P., Dowse, H. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Resetting+the+Circadian+Clock+by+Social+Experience+in+Drosophila+melanogaster.">Resetting the Circadian Clock by Social Experience in Drosophila melanogaster.</a> Science. 298, 2010-2012 (2002).</li></ol>

在我们的设置，一个中等强度的空气脉冲瞬时停止飞行运动。当空气脉冲结束时，苍蝇被释放这种静止状态和locomote通常，如在图2所示。因为这空气脉冲可有效地同步运动的人口，我们用它来开始审判，因此，我们也可以研究运动的发病势头打破刺激。然而，该试剂盒可以做没有空气脉冲自启动后的运动参数是由空气脉冲（图2）的影响。 一个经常发生的问题与运动自动分析?...

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		<th>Comment</th>
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<tr>
<td>Square Chamber</td>
<td>Tool</td>
<td>Machine Shop</td>
<td>N/A</td>
<td>Design from Wolf et al. 20021</td>
</tr>
<tr>
<td>Digital Camera</td>
<td>Camera</td>
<td>Sharp</td>
<td>ViewcamZ VL-23</td>
<td>&nbsp;</td>
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<td>Flowmeter</td>
<td>Tool</td>
<td>Cole Parmer</td>
<td>SY-32003-12</td>
<td>&nbsp;</td>
<tr>
<td>Light Box</td>
<td>Tool</td>
<td>DNASTAR, Inc.</td>
<td>Seq-Easy</td>
<td>&nbsp;</td>
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<td>Charcoal Filter</td>
<td>Tool</td>
<td>Fisher Scientific</td>
<td>09-744-37</td>
<td>&nbsp;</td>
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<td>Soll technologies</td>
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high-resolution video tracking of locomotion in adult drosophila melanogaster

苍蝇提供了一个复杂的行为，由于在这一领域的研究人员提供的遗传工具的大量研究的重要模式。在学习的运动行为<em&gt;果蝇</em依赖的能力，能够在运动过程中或在一个给定的任务量化的变化。出于这个原因，一个高清晰度视频跟踪系统，如我们在本文中介绍的一个，是一个有价值的工具，在实时测量运动。我们的协议涉及的初始空气脉冲打破苍蝇的势头，在一个正方形室由30第二拍摄期间使用。一个跟踪程序是用来计算每个飞行的瞬时速度在10毫秒增量室。分析软件，然后编译这些数据，并输出各种参数，如平均速度，最大速度，在运动中所花费的时间，加速等，此协议将讨论行为的任务苍蝇正确的喂养和管理，处理无需麻醉或固定苍蝇，建立一个控制的环境中，和运行检测，从开始到结束。

Watch this Scientific Journal Video about High-Resolution Video Tracking of Locomotion in Adult Drosophila Melanogaster at JoVE.com

High-Resolution Video Tracking of Locomotion in Adult Drosophila Melanogaster

复杂的运动器官的行为在研究<em&gt;果蝇</em&gt;是依赖于量化在一个给定的飞行的运动变化的能力。本文演示了如何做到这一点使用了高分辨率的跟踪系统。

高分辨率视频跟踪移动的，在成年果蝇。

Flies provide an important model for studying complex behavior due to the plethora of genetic tools available to researchers in this field.  Studying ...

high-resolution-video-tracking-locomotion-adult-drosophila

Research

JoVE Journal

Biology

13.4K 次观看.  Brandeis. 复杂的运动器官的行为在研究果蝇是依赖于量化在一个给定的飞行的运动变化的能力。本文演示了如何做到这一点使用了高分辨率的跟踪系统。

视频: High-Resolution Video Tracking of Locomotion in Adult Drosophila Melanogaster

Dissection of Larval CNS in Drosophila Melanogaster

The central nervous system (CNS) of Drosophila larvae is complex and poorly understood.  One way to investigate the CNS is to use immunohistochemistry to examine the expression of various novel and marker proteins.  Staining of whole larvae is impractical because the tough cuticle prevents antibodies from penetrating inside the body cavity.  In order to stain these tissues it is necessary to dissect the animal prior to fixing and staining.  In this article we demonstrate how to dissect Drosophila larvae without damaging the CNS.  Begin by tearing the larva in half with a pair of fine forceps, and then turn the cuticle "inside-out" to expose the CNS.  If the dissection is performed carefully the CNS will remain attached to the cuticle.  We usually keep the CNS attached to the cuticle throughout the fixation and staining steps, and only completely remove the CNS from the cuticle just prior to mounting the samples on glass slides.  We also show some representative images of a larval CNS stained with Eve, a transcription factor expressed in a subset of neurons in the CNS.  The article concludes with a discussion of some of the practical uses of this technique and the potential difficulties that may arise.

In this article we demonstrate how to dissect the central nervous system from third instar Drosophila larvae.

在果蝇幼虫中枢神经系统的剖析

The central nervous system (CNS) of Drosophila larvae is complex and poorly understood.  One way to investigate the CNS is to use immunohistochemistry to ...

科研

生物学

High-resolution Measurement of Odor-Driven Behavior in Drosophila Larvae

Olfactory responses in Drosophila larvae have been traditionally studied in Petri dishes comprising a single peripheral odor source. In this behavioral paradigm, the experimenter usually assumes that the rapid diffusion of odorant molecules from the source leads to the creation of a stable gradient in the dish. To establish a quantitative correlation between sensory inputs and behavioral responses, it is necessary to achieve a more thorough characterization of the odorant stimulus conditions. In this video article, we describe a new method allowing the construction of odorant gradients with stable and controllable geometries. We briefly illustrate how these gradients can be used to screen for olfactory defects (full and partial anosmia) and to study more subtle features of chemotaxis behavior.

In this video article, we describe a new method allowing the construction of odorant gradients with stable and controllable geometries. We briefly illustrate how these gradients can be used to screen for olfactory defects (full and partial anosmia) and to study more subtle features of chemotaxis behavior.

高分辨率测量的气味驱动在果蝇幼虫的行为

Olfactory responses in Drosophila larvae have been traditionally studied in Petri dishes comprising a single peripheral odor source. In this behavioral ...

Visually Mediated Odor Tracking During Flight in Drosophila

Flying insects use visual cues to stabilize their heading in a wind stream. Many animals additionally track odors carried in the wind. As such, visual stabilization of upwind tracking directly aids in odor tracking. But do olfactory signals directly influence visual tracking behavior independently from wind cues? Additionally, recent advances in olfactory molecular genetics and neurophysiology have motivated novel quantitative behavioral analyses to assess the behavioral influence of (e.g.) genetically inactivating specific olfactory activation circuits. We modified a magnetic tether system originally devised for vision experiments by equipping the arena with narrow laminar flow odor plumes. Here we focus on experiments that can be performed after a fly is tethered and is able to navigate in the magnetic arena. We show how to acquire video images optimized for measuring body angle, how to judge stable odor tracking, and we illustrate two experiments to examine the influence of visual cues on odor tracking.

Here we describe how to optimize the acquired video image for an olfactory magnetic-tether (OMT) apparatus. We also describe two sample experimental protocols for studying visuo-olfactory fusion.

在果蝇的飞行过程中可视介导的气味追踪

Flying insects use visual cues to stabilize their heading in a wind stream. Many animals additionally track odors carried in the wind. As such, visual ...

Dissection of Oenocytes from Adult Drosophila melanogaster

In Drosophila melanogaster, as in other insects, a waxy layer on the outer surface of the cuticle, composed primarily of hydrocarbon compounds, provides protection against desiccation and other environmental challenges. Several of these cuticular hydrocarbon (CHC) compounds also function as semiochemical signals, and as such mediate pheromonal communications between members of the same species, or in some instances between different species, and influence behavior. Specialized cells referred to as oenocytes are regarded as the primary site for CHC synthesis. However, relatively little is known regarding the involvement of the oenocytes in the regulation of the biosynthetic, transport, and deposition pathways contributing to CHC output. Given the significant role that CHCs play in several aspects of insect biology, including chemical communication, desiccation resistance, and immunity, it is important to gain a greater understanding of the molecular and genetic regulation of CHC production within these specialized cells. The adult oenocytes of D. melanogaster are located within the abdominal integument, and are metamerically arrayed in ribbon-like clusters radiating along the inner cuticular surface of each abdominal segment. In this video article we demonstrate a dissection technique used for the preparation of oenocytes from adult D. melanogaster. Specifically, we provide a detailed step-by-step demonstration of (1) how to fillet prepare an adult Drosophila abdomen, (2) how to identify the oenocytes and discern them from other tissues, and (3) how to remove intact oenocyte clusters from the abdominal integument. A brief experimental illustration of how this preparation can be used to examine the expression of genes involved in hydrocarbon synthesis is included. The dissected preparation demonstrated herein will allow for the detailed molecular and genetic analysis of oenocyte function in the adult fruit fly.

Dissection of Oenocytes from Adult Drosophila melanogaster

In insects, the oenocytes produce cuticular hydrocarbon compounds. These compounds protect against desiccation and facilitate chemical communication. Here we demonstrate a dissection technique used to isolate the oenocytes from adult Drosophila melanogaster, and illustrate how this preparation can be utilized to study genes involved in hydrocarbon synthesis.

从成人Oenocytes剖析果蝇</em

In Drosophila melanogaster, as in other insects, a waxy layer on the outer surface of the cuticle, composed primarily of hydrocarbon compounds, provides ...

Isolation of Drosophila melanogaster Testes

The testes of Drosophila melanogaster provide an important model for the study of stem cell maintenance and differentiation, meiosis, and soma-germline interactions. Testes are typically isolated from adult males 0-3 days after eclosion from the pupal case. The testes of wild-type flies are easily distinguished from other tissues because they are yellow, but the testes of white mutant flies, a common genetic background for laboratory experiments are similar in both shape and color to the fly gut. Performing dissection on a glass microscope slide with a black background makes identifying the testes considerably easier. Testes are removed from the flies using dissecting needles. Compared to protocols that use forceps for testes dissection, our method is far quicker, allowing a well-practiced individual to dissect testes from 200-300 wild-type flies per hour, yielding 400-600 testes. Testes from white flies or from mutants that reduce testes size are harder to dissect and typically yield 200-400 testes per hour.

Isolation of Drosophila melanogaster Testes

Drosophila melanogaster testes can be rapidly and efficiently isolated from adult males using dissecting needles. With practice, one can readily isolate in one or two days an amount of testes sufficient for the analysis of DNA or RNA by high throughput sequencing or more traditional molecular biology methods or of protein for antibody- or enzyme-based assays.

分离果蝇睾丸

The testes of Drosophila melanogaster provide an important model for the study of stem cell maintenance and differentiation, meiosis, and soma-germline ...

Insulin Injection and Hemolymph Extraction to Measure Insulin Sensitivity in Adult Drosophila melanogaster

Conserved nutrient sensing mechanisms exist between mammal and fruit fly where peptides resembling mammalian insulin and glucagon, respectively function to maintain glucose homeostasis during developmental larval stages 1,2. Studies on largely post-mitotic adult flies have revealed perturbation of glucose homeostasis as the result of genetic ablation of insulin-like peptide (ILP) producing cells (IPCs) 3. Thus, adult fruit flies hold great promise as a suitable genetic model system for metabolic disorders including type II diabetes. To further develop the fruit fly system, comparable physiological assays used to measure glucose tolerance and insulin sensitivity in mammals must be established. To this end, we have recently described a novel procedure for measuring oral glucose tolerance response in the adult fly and demonstrated the importance of adult IPCs in maintaining glucose homeostasis 4,5. Here, we have modified a previously described procedure for insulin injection 6 and combined it with a novel hemolymph extraction method to measure peripheral insulin sensitivity in the adult fly. Uniquely, our protocol allows direct physiological measurements of the adult fly's ability to dispose of a peripheral glucose load upon insulin injection, a methodology that makes it feasible to characterize insulin signaling mutants and potential interventions affecting glucose tolerance and insulin sensitivity in the adult fly.

Insulin Injection and Hemolymph Extraction to Measure Insulin Sensitivity in Adult Drosophila melanogaster

Conserved insulin signaling pathways found in the fruit fly Drosophila melanogaster make this organism a potential tool for modeling metabolic disorders including type II diabetes. To this end, it is critical to establish physiological assays to effectively measure systemic insulin action in peripheral glucose disposal in the adult fly.

注射胰岛素和血淋巴的提取来衡量成人胰岛素敏感性果蝇</em

Conserved nutrient sensing mechanisms exist between mammal and fruit fly where peptides resembling mammalian insulin and glucagon, respectively function ...

Measurement of Lifespan in Drosophila melanogaster

Aging is a phenomenon that results in steady physiological deterioration in nearly all organisms in which it has been examined, leading to reduced physical performance and increased risk of disease. Individual aging is manifest at the population level as an increase in age-dependent mortality, which is often measured in the laboratory by observing lifespan in large cohorts of age-matched individuals. Experiments that seek to quantify the extent to which genetic or environmental manipulations impact lifespan in simple model organisms have been remarkably successful for understanding the aspects of aging that are conserved across taxa and for inspiring new strategies for extending lifespan and preventing age-associated disease in mammals.
The vinegar fly, Drosophila melanogaster, is an attractive model organism for studying the mechanisms of aging due to its relatively short lifespan, convenient husbandry, and facile genetics. However, demographic measures of aging, including age-specific survival and mortality, are extraordinarily susceptible to even minor variations in experimental design and environment, and the maintenance of strict laboratory practices for the duration of aging experiments is required. These considerations, together with the need to practice careful control of genetic background, are essential for generating robust measurements. Indeed, there are many notable controversies surrounding inference from longevity experiments in yeast, worms, flies and mice that have been traced to environmental or genetic artifacts1-4. In this protocol, we describe a set of procedures that have been optimized over many years of measuring longevity in Drosophila using laboratory vials. We also describe the use of the dLife software, which was developed by our laboratory and is available for download (<a href="http://sitemaker.umich.edu/pletcherlab/software">http://sitemaker.umich.edu/pletcherlab/software</a>). dLife accelerates throughput and promotes good practices by incorporating optimal experimental design, simplifying fly handling and data collection, and standardizing data analysis. We will also discuss the many potential pitfalls in the design, collection, and interpretation of lifespan data, and we provide steps to avoid these dangers.

Measurement of Lifespan in Drosophila melanogaster

Drosophila melanogaster is a powerful model organism for exploring the molecular basis of longevity regulation. This protocol will discuss the steps involved in generating a reproducible, population-based measurement of longevity as well as potential pitfalls and how to avoid them.

寿命测量<em&gt;果蝇</em

Aging is a phenomenon that results in steady physiological deterioration in nearly all organisms in which it has been examined, leading to reduced ...

Maintenance of a Drosophila melanogaster Population Cage

Large quantities of DNA, RNA, proteins and other cellular components are often required for biochemistry and molecular biology experiments. The short life cycle of Drosophila enables collection of large quantities of material from embryos, larvae, pupae and adult flies, in a synchronized way, at a low economic cost. A major strategy for propagating large numbers of flies is the use of a fly population cage. This useful and common tool in the Drososphila community is an efficient way to regularly produce milligrams to tens of grams of embryos, depending on uniformity of developmental stage desired. While a population cage can be time consuming to set up, maintaining a cage over months takes much less time and enables rapid collection of biological material in a short period. This paper describes a detailed and flexible protocol for the maintenance of a Drosophila melanogaster population cage, starting with 1.5 g of harvested material from the previous cycle.

Maintenance of a Drosophila melanogaster Population Cage

This manuscript reports a detailed protocol for culturing, on a regular basis, a population of Drosophila melanogaster using a fly population cage.

的维持<em&gt;果蝇</em&gt;人口笼

Large quantities of DNA, RNA, proteins and other cellular components are often required for biochemistry and molecular biology experiments. The short life ...

Optical Cross-Sectional Muscle Area Determination of Drosophila Melanogaster Adult Indirect Flight Muscles

Muscle mass wasting, known as muscle atrophy, is a common phenotype in Drosophila models of neuromuscular diseases. We have used the indirect flight muscles (IFMs) of flies, specifically the dorso-longitudinal muscles (DLM), as the experimental subject to measure the atrophic phenotype brought about by different genetic causes. In this protocol, we describe how to embed fly thorax muscles for semi thin sectioning, how to obtain a good contrast between muscle and the surrounding tissue, and how to process optical microscope images for semiautomatic acquisition of quantifiable data and analysis. We describe three specific applications of the methodological pipeline. First, we show how the method can be applied to quantify muscle degeneration in a myotonic dystrophy fly model; second, measurement of muscle cross-sectional area can help to identify genes that either promote or prevent muscle atrophy and/or muscle degeneration; third, this protocol can be applied to determine whether a candidate compound is able to significantly modify a given atrophic phenotype induced by a disease-causing mutation or by an environmental trigger.

Optical Cross-Sectional Muscle Area Determination of Drosophila Melanogaster Adult Indirect Flight Muscles

We report a method to quantify muscle area, which is an indirect method to determine muscle mass in Drosophila adults. We demonstrate the application of our methodology by analyzing the indirect flight muscles in a Drosophila model of Myotonic Dystrophy disease.

果蝇成人间接飞行肌的光学横截肌面积测定

Muscle mass wasting, known as muscle atrophy, is a common phenotype in Drosophila models of neuromuscular diseases. We have used the indirect flight ...

Oral Bacterial Infection and Shedding in Drosophila melanogaster

The fruit fly Drosophila melanogaster is one of the best developed model systems of infection and innate immunity. While most work has focused on systemic infections, there has been a recent increase of interest in the mechanisms of gut immunocompetence to pathogens, which require methods to orally infect flies. Here we present a protocol to orally expose individual flies to an opportunistic bacterial pathogen (Pseudomonas aeruginosa) and a natural bacterial pathogen of D. melanogaster (Pseudomonas entomophila). The goal of this protocol is to provide a robust method to expose male and female flies to these pathogens. We provide representative results showing survival phenotypes, microbe loads, and bacterial shedding, which is relevant for the study of heterogeneity in pathogen transmission. Finally, we confirm that Dcy mutants (lacking the protective peritrophic matrix in the gut epithelium) and Relish mutants (lacking a functional immune deficiency (IMD) pathway), show increased susceptibility to bacterial oral infection. This protocol, therefore, describes a robust method to infect flies using the oral route of infection, which can be extended to the study of a variety genetic and environmental sources of variation in gut infection outcomes and bacterial transmission.

Oral Bacterial Infection and Shedding in Drosophila melanogaster

This protocol describes methods to orally expose and infect the fruit fly Drosophila melanogaster with bacterial pathogens, and to measure the number of infectious bacteria shed following gut infection. We further describe the effect of immune mutants on fly survival following oral bacterial infection.

果蝇的口腔细菌感染和脱落

The fruit fly Drosophila melanogaster is one of the best developed model systems of infection and innate immunity. While most work has focused on systemic ...