Name: the treadwheel: interval training protocol for gently induced exercise in drosophila melanogaster
Uploaded: 2018-06-08T15:00:00.000+00:00
Duration: 7 min 21 s
Description: Watch this Scientific Journal Video about The TreadWheel: Interval Training Protocol for Gently Induced Exercise in Drosophila melanogaster at JoVE.com

我们要感谢朱莉 Jarnigan, 梅瑞狄斯欧文斯, 雷切尔希尔, 布兰登上莫邪, 劳拉 Mafla, 奥利维亚鱼, 其余的芦苇实验室帮助他们的畜牧和图像处理。TreadWheel 在 UAB 机器店的协助下建造了原始的建筑。这项研究的经费由 NIH-R01 GM 098856 LKR 和阿拉巴马州大学的本科生创造力和研究学院提供给克尔苏加德和 BJW。

1. TreadWheel 设置和操作
注: 请参阅补充图 1 , 以了解工程机械结构示意图。所需零件列在材料表中。"TreadWheel"、"运动机器" 和 "机器" 这几句话在整个协议中交替使用。
<ol><li>
使用电源开关校准电机, 使整个旋转周期为十五年代 (4 rpm)。在将飞瓶放置到旋转臂上的固定夹具上之前, 校准机器转速。
	<ol>
<li>使用标准秒表和静止物体临时附着在机器上, 如用刷子触摸到马达盖上的小油漆刷, 只需触碰其中一个旋转托架即可调整速度。时间的旋转次数在一分钟内使用静止对象作为参考点, 并调整速度, 以达到十五年代旋转周期 (4 rpm)。</li>
	</ol>
</li>
</ol>2. 飞行收集和维护
注: 所有苍蝇都保持在25摄氏度孵化器与50% 的湿度和 12 h 光/暗循环之间的台式操作描述如下。除非另有说明, 苍蝇被喂食标准的玉米糖浆实验室饮食。
<ol><li>
用苹果汁琼脂板, 每盘一滴酵母膏, 和6盎司的塑料瓶, 为每个感兴趣的基因型准备产卵室。
	<ol>
<li>要准备苹果汁琼脂板, 填充35毫米 x 10 毫米培养皿¾-满3% 琼脂溶解到商店购买的苹果汁。在冰箱里储存固化的盘子, 直到使用。</li>
		<li>在50毫升锥形离心管中, 将商店购买的活性干酵母与蒸馏水相结合, 在每克酵母的比例为2毫升水。混合酵母与水, 以创建一个浆糊与番茄酱的一致性使用玻璃搅拌杆。根据需要调整酵母和水的比例以达到预期的一致性。</li>
		<li>将酵母膏贮存在冰箱中使用。保持管帽松动, 以允许消除毒气。</li>
		<li>用锋利的针或刀把小孔打进6盎司的方形底瓶 (聚丙烯), 作为铺设室的通风孔。</li>
		<li>在每个苹果汁琼脂板上涂一小点酵母膏 (直径约3毫米)。</li>
	</ol>
</li>
	<li>安置期望基因型的成人飞行入6盎司瓶和瓶盖瓶与苹果汁琼脂板材。把盘子固定在带橡胶带的瓶子上。反转, 并放置在孵化器允许成人产卵。</li>
	<li>每天两次 (早晚) 在铺设室更换苹果汁琼脂盘子。将用过的盘子放在孵化器中, 另12–24个 h, 让第一龄幼虫孵化。</li>
	<li>使用薄画笔或收集拾取 (附着在手柄上的挠性细针) 收集第一龄幼虫。将50个特定基因型的第一龄幼虫放置在所需实验食物类型的食物瓶中, 在孵化箱中发育和完成变形 (图 2a)。</li>
	<li>
收集成人进行运动治疗 (图 2A)。
	<ol>
<li>在蛹, 收集蛹与一个小, 潮湿的画笔和地方在空瓶之前, 羽化, 以防止成年苍蝇暴露在幼虫饮食。</li>
		<li>当他们 eclose 的时候, 把成年苍蝇转移到标准实验室饮食的食物瓶里。羽化后五天, 根据性别分开飞行。</li>
	</ol>
</li>
	<li>将收集到的成年人中的一半指定给实验和对照组。在每瓶50只苍蝇的标准实验室饮食中保持苍蝇的含量。</li>
	<li>每两天将成年人转移到新鲜食品瓶中, 以防止他们在运动中被卡住。在介绍苍蝇之前, 立即在新食物中添加几粒补充活酵母。 注意: 虽然有可能保持苍蝇没有活酵母补充, 我们发现, 成年苍蝇表现最好的补充。活酵母补充是一个变量, 可以修改, 以适应研究者的实验目标。</li>
</ol>3. 练习协议
<ol><li>在运动的第一天 (1 天), 推瓶插头离开1厘米的空间控制瓶和6厘米的空间之间的食物和插头的运动瓶 (图 1C)。 注: 控制瓶将有有限的空间, 苍蝇在运动机器上移动, 因此苍蝇将从事比运动小瓶更少的运动。</li>
	<li>把小瓶放在夹子里, 然后给苍蝇10分钟在机器上适应 (图 1B)。 注: 另一种控制是保持未动用苍蝇在台式旁边的运动机与6厘米的空间 (在瓶子), 以正常移动。与 "TreadWheel 1 厘米" 和 "6 厘米台式" 控制有关的运动的相似结果已被发现11。</li>
	<li>
在连续5天的旋转和休息交替回合 (图 1D) 的情况下, 将控制和运动苍蝇以恒定的室温旋转到机器上。 注意: 要避免温度对活动水平的混杂影响, 需要恒温。对于温度操纵研究, 将其放在孵化器内, 修改运动训练过程中的温度处理。<ol>
<li>运行这台机器四15分钟的运动在1天。交替这些回合与5分钟休息期。</li>
		<li>接下来的每一天, 增加5分钟的锻炼到一个已经存在的运动回合。例如, 在2天, 使第一个练习回合20分钟长, 但保持其余的回合在15分钟长。</li>
		<li>在3天, 运动飞行20分钟在前两次比赛和15分钟为以下两次回合。</li>
		<li>在4天, 除最后一回合外, 使用20分钟的运动比赛, 这应该是15分钟长。</li>
		<li>在5天, 保持所有运动回合20分钟长。 注: 在休息期间, 含有苍蝇的瓶子应保持牢固地夹紧。在这些时期, 瓶子的方向是不相关的, 因为所有苍蝇所经历的引力将来自同一个方向。</li>
	</ol>
</li>
	<li>在完成了某一天的运动治疗后, 将食物瓶插头放到正常位置, 并返回到第二天的孵化箱。</li>
	<li>在完整的锻炼制度完成后, 麻醉飞行使用标准飞行 CO2麻醉方法。然后将苍蝇转移到新的食品瓶或离心管, 以评估感兴趣的表型特征。 注: 例子表型包括攀登能力 (食物小瓶), 甘油三酯存储 (离心管), 或基因表达 (离心管)。该协议可以在这里暂停, 并根据要执行的评估适当存储样本。涉及活蝇的化验将需要按照表型化验的具体目标迅速进行。</li>
</ol>4. 攀登评估
<ol><li>
在练习协议完成后的一天休息后, 通过快速迭代负 geotaxis (环) 像阴性 geotaxis 攀登试验22测试一般的飞行攀爬性能。
	<ol>
<li>在空瓶子中放置10只苍蝇, 使用 CO2麻醉 (或直接从食品瓶中攻下), 用石蜡膜密封以防止苍蝇逃逸。给苍蝇至少10分钟适应新瓶。为了防止瓶子打翻, 胶带5厘米直径卡片圆到瓶子底部提供额外的表面面积。 注: 不应使用玻璃瓶插头, 因为它们阻碍了用于确定高度攀升的瓶子的一部分。</li>
		<li>放置在一个1厘米 x 1 厘米网格前面的小瓶20厘米在一个安装的, 固定的摄像头与内置定时器 (智能手机与定时器相机应用程序的工作很好)。</li>
		<li>轻轻地在柜台上轻敲小瓶三次, 把所有苍蝇敲到瓶子的底部。第三次攻下小瓶后, 立即启动 4 s 相机定时器捕捉图像, 以确定每只苍蝇爬的高度。 注: 轻拍应在一致的强度和相同的研究员在每个演习控制对比, 以尽量减少变化的苍蝇的反应, 由于攻牙。</li>
		<li>重复步骤4.1.3 两次, 在试验中至少休息1分钟, 每瓶共产生三项试验。</li>
	</ol>
</li>
	<li>
为了保存苍蝇的其他分析, 立即在攀登化验完成后, 闪光冻结苍蝇浸泡在液氮。
	<ol>
<li>使用一个浅, 开放顶部的杜瓦瓶填充到深度约5厘米与液态氮。</li>
		<li>使用 CO2麻醉将苍蝇转到小离心管, 然后将管子放在装满液氮的杜瓦瓶中。或者, 如果未来的分析需要避免 CO2, 快照冻结的苍蝇直接在他们的上升瓶通过轻轻拍打苍蝇到他们的瓶子底部, 而底部被淹没在液态氮, 然后转移冰冻苍蝇离心管使用镊子。 注: 液氮是低温的, 应仅用于适当的保护设备和房间通风。</li>
	</ol>
</li>
	<li>
使用 ImageJ23多点选择工具处理图像。
	<ol>
<li>在 ImageJ 中打开图像进行处理。</li>
		<li>根据背景网格纸设置一个1厘米的刻度, 计算每个单独飞行所爬的距离。使用工具栏上的 "线条工具" 从背景网格纸上跟踪1厘米2平方的一侧。单击 "分析" 选项卡并选择 "设置比例"。将 "已知距离" 设置为 ' 1.00 ' 和 "长度单位" 作为 "cm", 确保 "全局" 未选中, 然后单击 "确定"。</li>
		<li>从工具栏中选择 "多点选择" 图标, 然后放大图像中小瓶的底部。通过单击瓶子的最低部分, 将第一个点设置为瓶子的底部。</li>
		<li>单击瓶子中每只苍蝇的中心, 将其标记为数据点。请注意, 每瓶总共有11分, 一个标记底部的瓶子, 一个为每一个苍蝇。</li>
		<li>单击 "分析" 选项卡并选择 "度量" 以生成测量值表。将表保存为 csv 文件。</li>
		<li>在电子表格程序中打开 csv 文件, 然后通过从 "测量值" 表中的点1减去点2-11 的 y 值, 计算每次飞行所爬的精确距离。</li>
		<li>对每个图像重复步骤4.3.1 到4.3.6。</li>
	</ol>
</li>
</ol>5. 甘油三酯贮藏试验
注: 样品, 标准, 甘油标准溶液, 和 triglycerol 工作解决方案在整个测试中使用时应保持在冰上, 在该协议的有效期内, 应储存在冰箱时, 不使用。
<ol><li>
使用甘油标准溶液制备标准。
	<ol>
<li>将1000µL 均匀化缓冲器添加到标记为 b 的离心管中, 使空白 (b)。</li>
		<li>使标准 1 (S1), 2.5 毫克/毫升每井, 通过添加1000µL 的甘油标准溶液的离心管标记 S1。</li>
		<li>标准 2 (S2), 1.25 毫克/毫升每井, 通过添加500µL 的甘油标准溶液500µL 蝶和2O 在离心管标记 S2。</li>
		<li>标准 3 (S3), 0.625 毫克/毫升每井, 通过添加250µL 的甘油标准溶液750µL 蝶和2O 在离心管标记 S3。</li>
		<li>标准 4 (S4), 0.3125 毫克/毫升每井, 通过添加125µL 的甘油标准溶液的875µL diH20 在离心管标记 S4。</li>
	</ol>
</li>
	<li>
从血清甘油三酯测定试剂盒中制备 triglycerol 工作液。
	<ol>
<li>添加40毫升的蝶和2O 的免费甘油试剂从试剂盒和混合通过反转瓶。</li>
		<li>从试剂盒中加入10毫升的蝶和2O 到甘油三酯溶液, 并通过倒置瓶混合。</li>
		<li>结合甘油试剂和甘油三酯溶液, 并通过反转混合, 准备 triglycerol 工作溶液。 注: triglycerol 工作解决方案是良好的60天, 并可以完成四96井板化验每套件使用下面的协议。所用的体积从官方血清甘油三酯测定试剂盒中按比例缩小, 用于96井板格式。</li>
	</ol>
</li>
	<li>
通过仔细吹打每个标准的5µL 到一个96井平底, 明确微板块三个 (或四份为空白), 验证空白和标准。
	<ol>
<li>将125µL 的 Triglycerol 工作解决方案添加到每个包含标准的井中。允许解决方案进行30分钟的反应。</li>
		<li>将96井板放在分光光度计中, 读 540 nm 的吸光度。使用吸光度测量来创建标准曲线并通过评估 R2值来验证标准的准确性。如果标准的 R2小于 0.98, 则更仔细地重新制定标准。</li>
	</ol>
</li>
	<li>
开始1天的检测甘油三酯存储14使用离心管含有10冷冻苍蝇每。 注: 重复或多通道吸管不应在此步骤中使用, 因为在化验中使用的每个试剂的数量必须非常准确和一致, 以获得可重现的结果。在我们的经验中, 重复和多通道吸管没有这个程度的准确性和一致性。每一个通道吸管的整除都可以由实验者在视觉上得到质量控制的证实, 任何单个通道的吸管特定的偏差都将被所有的样本所体验到相同的程度。<ol>
<li>在玻璃瓶中结合0.272 克的200毫升2PO4、400µL、0.5 米 EDTA 和199.6 毫升的蝶和2O, 制备均匀化缓冲液。</li>
		<li>将100µL 均匀化缓冲液添加到40离心管中, 其中包括10名成人在攀爬试验后冷冻。然后, 离心机样品三十年代在1.8万克。</li>
		<li>研磨苍蝇使用电动磨床和杵 (或其他形式的组织均匀化), 以准备一个乳白色的解决方案。确保每个样品都使用新的杵。然后, 离心机样品在1.8万克2分钟。</li>
		<li>吸管只有前75µL 的上清液从每管进入新的离心管。确保从颗粒中没有飞位转移过来。把新的管子放在冰箱里过夜。</li>
	</ol>
</li>
	<li>
从冰箱中除去1天的样品, 开始2天的甘油三酯协议。 注意: 步骤5.5 可以在同一天进行 5.4, 但是我们发现在冰箱里过夜后甘油三酯浓度的信号会更加健壮。但是, 步骤5.4 和5.5 之间的时间不应超过36小时。<ol>
<li>选涡流第一管与上清液十年代和吸管5µL 成一个干净的离心管。添加95µL 0.15 米氯化钠进入新的离心管。存储这些标本在-20 °c 冷藏库为以后蛋白质化验如果需要。 注: 这些样品的上清与添加 NaCl 溶液可以用来确定蛋白质含量使用研究员的首选化验。我们使用布拉德福德方法24。蛋白质含量是衡量甘油三酯含量的一种方法, 但研究者在解释这些比值时也应谨慎, 因为运动和饮食也会影响蛋白质的贮存。</li>
		<li>对于原上清液瓶, 涡流第一管与上清液体十年代, 然后吸管5µL 进入两个单独的油井96井微板块, 以产生技术复制。一定要注意每个样品使用的井。重复这一步骤, 为39其他管。</li>
		<li>吸管确切地5µL 的空白入微板块的四个井, 并且为剩余的井, 吸管确切地5µL 每标准在三个。</li>
		<li>为每个井添加精确的125µL triglycerol 工作解决方案, 并允许解决方案进行30分钟的反应。</li>
		<li>将96井板放在分光光度计中, 读 540 nm 的吸光度。</li>
		<li>使用从标准计算的标准曲线将吸光度值转换为浓度 (毫克/毫升)。</li>
	</ol>
</li>
</ol>

<ol>
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作者没有什么可透露的。

这里详细的练习协议已经证明可以成功地刺激果蝇的温和运动, 并且可以用来模拟在控制的实验室设置11的耐力运动。应该指出的是, 在最初开发 TreadWheel 概念时, 我们考虑了可能被修改以执行类似的商业产品 (例如实验室烤)。然而, 我们最终拒绝了这种方法, 选择了定制的设计, 因为商用设备没有足够低的转速 (4 rpm), 并缺乏足够的小瓶容量的高通量样品生成。
该协议本身可以进行调整, 以涵盖广泛的研究课题。例如, 调整苍蝇的运动机制的频率和持续时间可以改变他们接受的工作的强度。通过延长该议定书长于一周或其他年龄组, 就有可能研究运动对衰老和各种年龄相关疾病的影响。我们建议, 如果实现了扩展的练习方法, 则应至少添加一周的休息日以允许恢复。在包括恢复日的制度中, 表现和改进比使用连续每日练习12的制度更大。此外, 我们会告诫不要延长运动的长度超过30分钟, 因为适应旋转运动可以发生后长时间连续运动11。我们也观察到苍蝇的动机的遗传变异, 以维持其活动水平在11。因此, 如果需要更长的回合, 考虑监测苍蝇的活动水平, 如渡边和谜语15,21描述他们修改的总概念。其他可能变化的因素包括温度和睡眠-唤醒模式, 这也被证明也影响飞行运动26,27。
在协议完成期间和之后有大量的可用分析来解决运动对生理、行为和分子水平的影响, 以及解开运动的后生效应的潜力。额外的新陈代谢化验 (如葡萄糖, 糖原和蛋白质), 心脏表现和炎症反应措施可以用来进一步探索效果锻炼对飞行生理学和身体构成12,14 ,24,28。与运动诱导相关的各种行为变化, 如喂养行为、运动活动和睡眠的变化, 也可以用诸如咖啡馆化验29或活动监测设备12、15 等工具来测量. ,30。运动的基因表达和细胞呼吸变化也可以用 qRT PCR11和 respirometry31等方法进行量化。最后,果蝇有宝贵的遗传资源, 如果蝇遗传学参考面板2和果蝇合成人口资源, 为研究人员提供平台来执行定量遗传学研究25,32。这些工具使制图实验, 如全基因组的关联研究和数量性状基因座映射, 以确定与运动和饮食相关的候选基因座。
使用 TreadWheel 完成的研究表明, 平均而言, 运动能降低体重, 总甘油三酯储存, 糖原, 同时增加蛋白质含量和攀登性能11。此外, 在性别和基因型的体重, 以及甘油三酯, 蛋白质, 糖原, 葡萄糖和活动水平11,15有变数的反应。虽然在基因型和性别上对运动和饮食的反应变化可能具有挑战性, 但有时, 它反映了自然种群中观察到的实际生物变异的来源。当我们努力了解导致大都会高比率的潜在因素的多样性时, 通过测试模型有机体中的这些因素来促进解析各种因素的相对作用的工具将对我们的能力至关重要。制定个性化的预防和治疗干预措施。要充分评估运动的有效性, 在进行实验和制定结论时, 必须考虑每个因素及其相互作用。
与大多数其他的飞行运动机器一样, 它的量化飞行运动的能力有限。最近, 渡边和谜语开发了旋转运动定量系统 (满足要求), 一个从果蝇监测系统系列15,21的活动监测单元 (LAM25H) 修改的一组主干骨干。与 LAM25H 一样, 这个系统使用旋转运动来轻轻地诱发运动, 同时使用该臂来跟踪和量化飞行活动, 但它持有的小瓶数量 (32 瓶) 比15。对于高通量的运动研究, 如果没有必要或不需要活动量化, 那么它就可以获得更大的样本数。总的设计也可以从目前的形式修改, 以容纳更多的瓶子。这个系统, 连同其他现有的锻炼方法, 帮助确定运动不仅可以在果蝇中实现, 而且还可研究以确定物理活动对各种生理和分子反应的影响。11,12,15。因此, 作为一种行之有效的方法来诱导苍蝇的温和运动, 它可以用来探索大量的生物问题。

儿童和成人肥胖症在文化中日益流行, 它们消耗高热量的饮食, 长期闲置, 这会导致严重的远期后果, 包括胰岛素抵抗、慢性炎症和骨关节炎。1,2,3,4. 由于脂肪和糖的消费量增加以及主要的生活习惯5, 这些疾病的流行率继续上升, 原因是热量摄入和支出的不平衡日益严重。相应地, 这种能量失衡导致2型糖尿病 (T2DM) 和心血管疾病 (CVD)5的病例增加。如果患者被诊断患有紊乱代谢综合征 (大都会), 其症状包括腹部肥胖和血脂异常2, 个体就有更高的风险发展两种疾病。大都会是由基因型和各种环境因素, 如饮食和运动6的复杂相互作用所形成的。因此, 为了充分了解这一复杂疾病的基本机制, 所有这些因素都应加以考虑。
当谈到打击大都会时, 医生首先建议接受生活方式的改变, 包括饮食健康, 均衡饮食和身体活动2,7,8。由于有效的药物是有限的, 胃旁路手术费用高昂, 需要终生的医疗监测, 药理和外科干预只建议在严重的情况下, 只有结合这些生活方式的变化3 ,7,8。虽然生活方式干预, 如运动和节食, 可以产生和维持长期减肥目标, 无论这些纠正措施是否能充分改善与该研究的负面影响需要进一步学习7, 8。
老鼠模型被用来检查运动对新陈代谢疾病的影响多年;然而, 引进运动的飞行研究大都会是一个相对最近的努力9,10,11,12。苍蝇提供了一个完美的工具来研究运动在一个受控实验室设置, 因为它们易于操作, 寿命短, 维护成本低廉, 与能量相关的代谢通路高度保守的果蝇之间并且人13。果蝇基因组具有良好的特点, 有丰富的遗传学工具可用于果蝇, 可以提供洞察各种基因型和环境基因型的相互作用, 可以调节运动对生物体健康的影响14。
目前的果蝇运动方法援引苍蝇的先天, 消极的 geotaxis 倾向, 行为本能向上攀登, 刺激在他们的外壳11,12,15 的攀登.动力塔是一种刺激苍蝇运动的方法, 系统地将飞罩垂直向上, 然后将它们放回工作台面, 有效地将苍蝇敲到瓶子的底部, 从而诱发其本能的负 geotaxis。12,16. 使用这台机器进行的实验表明, 运动对许多与年龄有关的疾病, 包括 CVD 和 T2DM, 是一个强有力的保护因素, 并促进健康的衰老12,17,18.具体来说, 他们表明, 锻炼可以减少苍蝇与年龄相关的移动性下降的开始, 并改善多年龄相关的症状, 如心脏表现和压力反应17,18。然而, 控制苍蝇从来没有放置在塔上显示更高的攀登得分比那些经历的机器, 暗示的力量, 重复下降可能会伤害苍蝇和影响移动12。这表明, 另一种诱导运动的方法, 不那么有力, 避免造成身体创伤, 将是一个有用的, 互补的方法, 对16的动力塔协议。
为了在果蝇中轻轻地诱发运动,我们开发了一种叫做 TreadWheel 的运动机器 (图 1).TreadWheel (从今往后缩写为总的) 旋转运动, 通过不断地重新定义瓶子的引力顶部来触发苍蝇天生的、消极的 geotaxis 倾向, 这反过来又刺激了苍蝇的攀爬。与其他方法不同的是, 它的旋转运动本质上是温和的, 这会减少可能产生的额外压力源的数量, 并影响结果。因此, 该机器提供了方法来诱导大量的苍蝇运动, 而不诱发压力, 这将使研究人员研究运动对新陈代谢健康的影响 (图 2), 衰老, 睡眠, 以及许多其他主题11。
我们的方法遵循了一个反金字塔, 间隔训练协议, 这主要包括一些方面的好氧间隔训练和一些耐力运动训练。在本议定书中, 标准的联谊会制度被改变, 以逐步增加每间隔五天期间的持续时间, 以促进耐力。比起持续适度的锻炼19,20, 在预防代谢综合征的常见危险因素方面, 特里比其他干预方法特别有用。然而, 习惯相对于动力塔的不利之处在于, 飞行速度越快, 对旋转运动的习惯, 就会使苍蝇的固有倾向变异, 从而复杂化对运动益处的解释。11渡边和谜语15及其互补文章21描述了这个限制的优雅解决方案。

<table><tbody><tr><td>&lt;strong&gt;Materials for TreadWheel Construction:&lt;/strong&gt;</td><td></td><td></td><td></td></tr><tr><td>Heavy Duty Vibration-Damping leveling Mount</td><td>McMaster-Carr</td><td>60855K71</td><td>Quantity: 4</td></tr><tr><td>Stainless Steel Ball Bearing</td><td>McMaster-Carr</td><td>57155K306</td><td>Quantity: 8</td></tr><tr><td>Plug-in Voltage Transformer (500MA, 120VAC input, 24 VAC output)</td><td>McMaster-Carr</td><td>70235K16</td><td>Quantity: 1</td></tr><tr><td>Compact Square-Face DC Gear motor</td><td>McMaster-Carr</td><td>6409K23</td><td>Quantity: 1</td></tr><tr><td>Tool Holder (clamps)</td><td>McMaster-Carr</td><td>1723A22</td><td>Quantity: 5 (10x)</td></tr><tr><td>12L14 Carbon Steel Tight-Tolerance Rod</td><td>McMaster-Carr</td><td>5227T24</td><td>Quantity: 1</td></tr><tr><td>Set Screw Shaft Collar</td><td>McMaster-Carr</td><td>6432K13</td><td>Quantity: 8</td></tr><tr><td>Round-Belt Pulley</td><td>McMaster-Carr</td><td>6284K51</td><td>Quantity: 5</td></tr><tr><td>Dart Controls &amp;ndash; 25 Max RPM, Electric AC DC Motor</td><td>McMaster-Carr</td><td>13DV 1A</td><td>Quantity: 1</td></tr><tr><td>&lt;strong&gt;Materials for Fly Maintenace and Husbandry&lt;/strong&gt;</td><td></td><td></td><td></td></tr><tr><td>6 oz Square Bottom Bottles (polypropylene)</td><td>Genesee Scientific</td><td>32-130</td><td>Quantity: 1</td></tr><tr><td>35x10mm Petri Dishes</td><td>VWR</td><td>82050-536</td><td>Quantity: 1</td></tr><tr><td>Narrow Drosophila vials</td><td>Genesee Scientific</td><td>32-116</td><td>Quantity: 1</td></tr><tr><td>Flystuff Flypad</td><td>Genesee Scientific</td><td>59-114</td><td>Quantity: 1</td></tr><tr><td>Blowgun, Mini</td><td>Genesee Scientific</td><td>54-104</td><td>Quantity: 1</td></tr><tr><td>&lt;strong&gt;Materials for RING-like Assay:&lt;/strong&gt;</td><td></td><td></td><td></td></tr><tr><td>ImageJ software</td><td>NIH</td><td>https://imagej.nih.gov/ij/</td><td>Quantity: 1</td></tr><tr><td>1 cM graph paper or drawn grid (at least 20 cM by 30 cM)</td><td>various</td><td></td><td>Quantity: 1</td></tr><tr><td>digital camera with timer or smart phone with camera timer app</td><td>various</td><td></td><td>Quantity: 1</td></tr><tr><td>&lt;strong&gt;Materials for Triglyceride Assay:&lt;/strong&gt;</td><td></td><td></td><td></td></tr><tr><td>Dewar Flask</td><td>VWR</td><td>14200-960</td><td>Quantity: 1</td></tr><tr><td>Serum Triglyceride Determination Kit</td><td>Sigma Aldrich</td><td>TRO100</td><td>Quantity: 1</td></tr><tr><td>Cordless Pestle Motor</td><td>VWR</td><td>47747-370</td><td>Quantity: 1</td></tr><tr><td>Pestles</td><td>VWR</td><td>47747-358</td><td>Quantity: 1</td></tr></tbody></table>

the treadwheel: interval training protocol for gently induced exercise in drosophila melanogaster

复杂的代谢疾病的发病率增加了, 由于广泛的转变为生活方式的增加卡路里摄入量和降低活动水平。这些多因素疾病产生于遗传, 环境和行为因素的结合。其中一种复杂的疾病是代谢综合征, 这是代谢紊乱的一组, 包括高血压、高血糖和腹部肥胖。运动和饮食干预是医生建议的主要治疗方法, 以减轻肥胖及其随后的代谢性疾病。运动干预, 特别是有氧间歇训练, 刺激2型糖尿病 (T2DM)、心血管疾病 (CVD) 和其他条件的共同危险因素的有利变化。随着大量证据的汇集, 描述了治疗效果锻炼对新陈代谢的健康, 建立一个系统, 模型运动在一个控制的设置提供了一个有价值的工具来评估运动的效果在实验环境中。果蝇是研究运动干预引起的生理和分子变化的一个很好的工具。与人类相比, 苍蝇有短寿命和类似的代谢养分机制。为了诱发果蝇的运动, 我们研制了一种叫 TreadWheel 的机器, 它利用苍蝇天生的、消极的 geotaxis 倾向来轻轻地诱发攀爬。这使研究人员能够对大量的基因型苍蝇进行实验, 以更好地了解运动对新陈代谢健康的影响所产生的基因分异的相互作用。

我们特别感兴趣的是找出有助于个人整体新陈代谢健康的因素。此前发现, 按饮食基因型相互作用对代谢性状的人口水平变化有很大贡献14。这意味着每个基因型都以独特和复杂的方式对环境差异作出反应。为了扩大我们对环境基因型效应的研究, 包括体育锻炼, 我们开发了 TreadWheel, 它能够以高通量的方式暴露大量的基因型到有氧间歇训练中。
为了确定运动是否影响了代谢特性, 我们测量了在俄勒冈州 R (不再回应买主) 和 y1w1苍蝇, 常见的 wildtype 苍蝇 (图 2B–C) 的甘油三酯存储, 并规范化这些值对苍蝇 '蛋白质浓度, 如最初报告在门德斯等。11. 我们通过多元方差分析 (MANOVA) 对基因型、性别、运动治疗 (及其相互作用) 进行了分析, 并对实验块效应 (如时间复制和食品瓶) 进行了研究, 发现有一个显著的由运动基因型相互作用 (p = 0.0017) 影响甘油三酯贮存。男性和女性之间存在显著的性二形效应, 男性储存的甘油三酯比雌性多 (p < 0.0001)。我们看到, 在雌性, 运动的苍蝇有明显降低甘油三酯水平比他们的未动用对应 (图 2B, p < 0.0001)。在男性中, 而在俄勒冈州 R 运动的苍蝇 (与对照组相比) 观察到的甘油三酯储存的减少没有统计学意义, 两个单独的线之间观察到甘油三酯储存的显著差异 (图2C, p < 0.0001)。请注意, 虽然标准化甘油三酯浓度对蛋白质浓度提供了洞察整体飞行身体成分比率, 比较直接甘油三酯或蛋白质浓度之间的不同群体的苍蝇也可以提供关于运动对这些表型的影响的具体信息。
由于诸如成人运动、性别和基因型等变量会影响甘油三酯的贮存, 预计这些因素也会影响到其他表位, 并与饮食互动。我们从一个代表性的 wildtype果蝇遗传参考面板线 (DGRP 153)25的幼虫, 在高脂肪或正常饮食和诱导运动的成人苍蝇一周 (图 2a)。随后, 我们进行了环形阴性 geotaxis 测定, 以测量攀爬能力。所用的攀爬试验与标准环试验不同;而不是一个环形装置, 用石蜡膜覆盖开口的小瓶被用来在试验期间盖住苍蝇。原环检测的其他方面, 如将苍蝇敲到瓶子底部并拍照的时间, 则保留了22。
所有治疗都重复了三次, 每一次至少有59个单独的苍蝇复制和治疗。MANOVA 对饮食、基因型、性别、运动治疗 (及其相互作用) 进行了统计分析, 并对时间复制、化验小瓶和小瓶化验复制等实验块效应进行了研究。我们发现, 运动的雌性在高脂肪饮食中比任何其他女性治疗 (图 2D) 上升了很高 (p < 0.005)。对于男性, 人们看到, 运动只提高了男性在正常饮食中的上升, 而在高脂肪饮食中上升的人显示没有显著的变化 (图 2E)。我们也发现了显著的性二形影响 (p < 0.0001) 在攀登与男性攀登高于女性。女性从 DGRP 153 线 (图 2D) 对女性消费正常饮食 (p < 0.0001) 的攀登表现下降的惊人结果是, 这种类型的运动如何可能不是一个例子对所有基因型有一致的积极干预, 可能取决于其他环境因素。在门德斯等。11, 女性从四其他基因线被测试为他们的上升的表现在正常膳食以后上升了所有显示了增强的攀登能力与锻炼训练。这表明在 DGRP 153 中观察到的反应是基因型特异性的, 而不是总的运动治疗的一般性质。不同性别、饮食和运动治疗组的反应差异表明, 有显著的按性别进行的交互运动, 影响了该线的攀爬能力 (p < 0.0001)。
结果表明, 运动对成年蝇代谢健康的影响可以是性别、基因型和幼虫饮食的功能。在其他研究11、12、14、15中也观察到了基因型、环境变量 (例如饮食和运动) 和性别的表观变异。因此, 大都会和果蝇可以成为一个强有力的策略来阐明遗传和环境因素, 形成代谢健康。
<img alt="Figure 1" class="xfigimg" src="/files/ftp_upload/57788/57788fig1.jpg"> 图 1: TreadWheel 运动机.(A) 该机器持有48瓶, 并具有可调的速度特性。实验以4转每分钟的转速进行。(B) 载有实验苍蝇的个别食物瓶被夹在一个旋转车轴上的托架上。(C) 瓶塞与食物之间的距离为运动瓶和1厘米的控制瓶6厘米。然后把瓶子放在机器上进行锻炼。(D) 5 天反金字塔运动制度被用来锻炼成年苍蝇。每天, 一个额外的五分钟被添加到一个练习回合增加锻炼强度递增, 模型耐力, 间歇训练。这个数字已经被修改了从门德斯等。11.<a href="https://www.jove.com/files/ftp_upload/57788/57788fig1large.jpg" target="_blank">请点击这里查看这个数字的更大版本.</a>
<img alt="Figure 2" class="xfigimg" src="/files/ftp_upload/57788/57788fig2.jpg"> 图 2: 视觉方法和代表性结果.(A) 检测幼虫饮食和成人运动交互作用的方法。幼虫在高脂肪或正常的实验室饮食中被饲养, 并在羽化后转为正常食物。成年苍蝇被性别分开, 被送入实验组, 连续五天锻炼。随后进行了环形阴性 geotaxis 攀登试验, 并对甘油三酯测量结果进行了冻结。代表甘油三酯数据显示两个基因线, 俄勒冈 R 和 y1w1 (B) 女性和 (C) 男性。所有苍蝇都是以正常的饮食饲养并在成年期内锻炼的。不同字母的水平有显著差异 (p < 0.05), 使用的是学生的t测试。这个数字包含了在门德斯等地报告的数据子集。11. 关于 (D) 女性和 (E) 男性的 DGRP 153 行显示了代表性攀登数据。每个点代表232个或更多个人在三个独立时间点的平均爬升性能。错误条指示一个标准错误。<a href="https://www.jove.com/files/ftp_upload/57788/57788fig2large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
<img alt="Supplemental Figure 1" src="/files/ftp_upload/57788/57788supfig1.jpg"> 补充图 1: TreadWheel 的详细示意图.(A) 用于将小瓶连接到机器上的双瓶钳结构示意图。(B) 机器旋转驱动系统的内部视图。(C) 机器正面的平行投射。<a href="https://www.jove.com/files/ftp_upload/57788/57788supfig1large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
补充代码文件: TreadWheel. skp<a href="https://www.jove.com/files/ftp_upload/57788/TreadWheel.skp" target="_blank">请单击此处下载此文件.</a>

Watch this Scientific Journal Video about The TreadWheel: Interval Training Protocol for Gently Induced Exercise in Drosophila melanogaster at JoVE.com

The TreadWheel: Interval Training Protocol for Gently Induced Exercise in Drosophila melanogaster

TreadWheel 利用旋转运动, 通过利用蝇的先天、负 geotaxis, 轻轻诱导成年果蝇运动。它允许分析运动和因素之间的相互作用, 例如基因型、性别和饮食, 以及它们对生理和分子检测的影响, 以评估代谢健康。

TreadWheel:果蝇轻度诱导运动的间歇训练协议

The incidence of complex metabolic diseases has increased as a result of a widespread transition towards lifestyles of increased caloric intake and ...

the-treadwheel-interval-training-protocol-for-gently-induced-exercise

Research

JoVE Journal

Genetics

The TreadWheel: Interval Training Protocol for Gently Induced Exercise in Drosophila melanogaster

11.6K 次观看.  University of Alabama. TreadWheel 利用旋转运动, 通过利用蝇的先天、负 geotaxis, 轻轻诱导成年果蝇运动。它允许分析运动和因素之间的相互作用, 例如基因型、性别和饮食, 以及它们对生理和分子检测的影响, 以评估代谢健康。

视频: The TreadWheel: Interval Training Protocol for Gently Induced Exercise in Drosophila melanogaster

Assaying Locomotor Activity to Study Circadian Rhythms and Sleep Parameters in Drosophila

Most life forms exhibit daily rhythms in cellular, physiological and behavioral phenomena that are driven by endogenous circadian (&equiv;24 hr) pacemakers or clocks. Malfunctions in the human circadian system are associated with numerous diseases or disorders. Much progress towards our understanding of the mechanisms underlying circadian rhythms has emerged from genetic screens whereby an easily measured behavioral rhythm is used as a read-out of clock function. Studies using Drosophila have made seminal contributions to our understanding of the cellular and biochemical bases underlying circadian rhythms. The standard circadian behavioral read-out measured in Drosophila is locomotor activity. In general, the monitoring system involves specially designed devices that can measure the locomotor movement of Drosophila. These devices are housed in environmentally controlled incubators located in a darkroom and are based on using the interruption of a beam of infrared light to record the locomotor activity of individual flies contained inside small tubes. When measured over many days, Drosophila exhibit daily cycles of activity and inactivity, a behavioral rhythm that is governed by the animal's endogenous circadian system. The overall procedure has been simplified with the advent of commercially available locomotor activity monitoring devices and the development of software programs for data analysis. We use the system from Trikinetics Inc., which is the procedure described here and is currently the most popular system used worldwide. More recently, the same monitoring devices have been used to study sleep behavior in Drosophila. Because the daily wake-sleep cycles of many flies can be measured simultaneously and only 1 to 2 weeks worth of continuous locomotor activity data is usually sufficient, this system is ideal for large-scale screens to identify Drosophila manifesting altered circadian or sleep properties.

Assaying Locomotor Activity to Study Circadian Rhythms and Sleep Parameters in Drosophila

We describe procedures for recording daily locomotor activity rhythms of Drosophila and subsequent data analysis. Locomotor activity rhythms are a reliable behavioral output of animal circadian clocks and are used as the standard readout of clock function when studying circadian mutants or examining how the environment regulates the circadian system.

含量自发活动，研究昼夜节律和睡眠参数果蝇</em

Most life forms exhibit daily rhythms in cellular, physiological and behavioral phenomena that are driven by endogenous circadian (&equiv;24 hr) ...

科研

神经科学

Measuring Exercise Levels in Drosophila melanogaster Using the Rotating Exercise Quantification System (REQS)

Drosophila melanogaster is a new model organism for studies in exercise biology. To date, two main exercise systems, the Power Tower and the Treadwheel have been described. However, a method to measure the amount of additional animal activity induced through the exercise treatment has been lacking. The Rotating Exercise Quantification System (REQS) fills this need, providing a measure of animal activity for animals experiencing rotational exercise. This protocol details how to use the REQS to assess animal activity during rotational exercise and illustrates the type of data that can be generated. Here, we demonstrate how the REQS is used to measure sex- and strain-specific differences in exercise induced activity. The REQS can also be used to evaluate the impact of various other experimental parameters such as age, diet, or population size on exercise induced activity. In addition, it can be used to compare the efficacy of different exercise training protocols. Importantly, it provides an opportunity to standardize exercise treatments between strains, allowing the researcher to achieve equal amounts of activity between groups if needed. Thus, the REQS is a notable new resource for exercise biologists working with the Drosophila model system and complements existing exercise systems.

Measuring Exercise Levels in Drosophila melanogaster Using the Rotating Exercise Quantification System REQS

The Rotating Exercise Quantification System (REQS) can induce exercise in Drosophila melanogaster through rotation while simultaneously measuring the amount of activity performed by the animals. Here, we present a point-by-point protocol detailing how to measure activity levels of animals experiencing rotational exercise treatments using the REQS.

使用旋转运动量化系统测量果蝇中的运动水平 (满足要求)

Drosophila melanogaster is a new model organism for studies in exercise biology. To date, two main exercise systems, the Power Tower and the Treadwheel ...

遗传学

High-Resolution Video Tracking of Locomotion in Adult Drosophila Melanogaster

Flies provide an important model for studying complex behavior due to the plethora of genetic tools available to researchers in this field. Studying locomotor behavior in Drosophila melanogaster relies on the ability to be able to quantify changes in motion during or in response to a given task. For this reason, a high-resolution video tracking system, such as the one we describe in this paper, is a valuable tool for measuring locomotion in real-time. Our protocol involves the use of an initial air pulse to break the flies momentum, followed by a thirty second filming period in a square chamber. A tracking program is then used to calculate the instantaneous speed of each fly within the chamber in 10 msec increments. Analysis software then compiles this data, and outputs a variety of parameters such as average speed, max speed, time spent in motion, acceleration, etc. This protocol will discuss proper feeding and management of flies for behavioral tasks, handling flies without anesthetization or immobilization, setting up a controlled environment, and running the assay from start to finish.

The study of complex locomotor behavior in Drosophila melanogaster is dependent upon the ability to quantify changes in a given fly's movement. This article demonstrates how to do this using a high-resolution tracking system.

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 ...

生物学

An Affordable and Efficient "Homemade" Platform for Drosophila Behavioral Studies, and an Accompanying Protocol for Larval Mitochondrial Respirometry

The usefulness of Drosophila as a model organism for the study of human diseases, behaviors and basic biology is unquestionable. Although practical, Drosophila research lacks popularity in developing countries, possibly due to the misinformed idea that establishing a lab and performing relevant experiments with such tiny insects is difficult and requires expensive, specialized apparatuses. Here, we describe how to build an affordable flylab to quantitatively analyze a myriad of behavioral parameters in D. melanogaster, by 3D-printing many of the necessary pieces of equipment. We provide protocols to build in-house vial racks, courtship arenas, apparatuses for locomotor assays, etc., to be used for general fly maintenance and to perform behavioral experiments using adult flies and larvae. We also provide protocols on how to use more sophisticated systems, such as a high resolution oxygraph, to measure mitochondrial oxygen consumption in larval samples, and show its association with behavioral changes in the larvae upon the xenotopic expression of the mitochondrial alternative oxidase (AOX). AOX increases larval activity and mitochondrial leak respiration, and accelerates development at low temperatures, which is consistent with a thermogenic role for the enzyme. We hope these protocols will inspire researchers, especially from developing countries, to use Drosophila to easily combine behavior and mitochondrial metabolism data, which may lead to information on genes and/or environmental conditions that may also regulate human physiology and disease states.

An Affordable and Efficient "Homemade" Platform for Drosophila Behavioral Studies, and an Accompanying Protocol for Larval Mitochondrial Respirometry

We provide protocols for anyone with a "maker culture" mind to start building a flylab for quantitative analysis of a myriad of behavioral parameters in Drosophila melanogaster, by 3D-printing many of the necessary pieces of equipment. We also describe a high resolution respirometry protocol using larvae to combine behavioral and mitochondrial metabolism data.

The usefulness of Drosophila as a model organism for the study of human diseases, behaviors and basic biology is unquestionable. Although practical, ...

行为学

A Simple Technique to Assay Locomotor Activity in Drosophila

Drosophila melanogaster is an ideal model organism for studying various diseases due to its abundance of advanced genetic manipulation techniques and diverse behavioral features. Identifying behavioral deficiency in animal models is a crucial measure of disease severity, for example, in neurodegenerative diseases where patients often experience impairments in motor function. However, with the availability of various systems to track and assess motor deficits in fly models, such as drug-treated or transgenic individuals, an economical and user-friendly system for precise evaluation from multiple angles is still lacking. A method based on the AnimalTracker application programming interface (API) is developed here, which is compatible with the Fiji image processing program, to systematically evaluate the movement activities of both adult and larval individuals from recorded video, thus allowing for the analysis of their tracking behavior. This method requires only a high-definition camera and a computer peripheral hardware integration to record and analyze behavior, making it an affordable and effective approach for screening fly models with transgenic or environmental behavioral deficiencies. Examples of behavioral tests using pharmacologically treated flies are given to show how the techniques can detect behavioral changes in both adult flies and larvae in a highly repeatable manner.

A Simple Technique to Assay Locomotor Activity in Drosophila

The present protocol assesses the locomotor activity of Drosophila by tracking and analyzing the movement of flies in a hand-made arena using open-source software Fiji, compatible with plugins to segment pixels of each frame based on high-definition video recording to calculate parameters of speed, distance, etc.

一种测定果蝇运动活性的简单技术

Drosophila melanogaster is an ideal model organism for studying various diseases due to its abundance of advanced genetic manipulation techniques and ...

Foraging Path-length Protocol for Drosophila melanogaster Larvae

The Drosophila melanogaster larval path-length phenotype is an established measure used to study the genetic and environmental contributions to behavioral variation. The larval path-length assay was developed to measure individual differences in foraging behavior that were later linked to the foraging gene. Larval path-length is an easily scored trait that facilitates the collection of large sample sizes, at minimal cost, for genetic screens. Here we provide a detailed description of the current protocol for the larval path-length assay first used by Sokolowski. The protocol details how to reproducibly handle test animals, perform the behavioral assay and analyze the data. An example of how the assay can be used to measure behavioral plasticity in response to environmental change, by manipulating feeding environment prior to performing the assay, is also provided. Finally, appropriate test design as well as environmental factors that can modify larval path-length such as food quality, developmental age and day effects are discussed.

Foraging Path-length Protocol for Drosophila melanogaster Larvae

We provide a detailed protocol for a Drosophila melanogaster foraging path-length assay. We discuss the preparation and handling of test animals, how to perform the assay and analyze the data.

觅食路径长度协议<em&gt;果蝇</em&gt;幼虫

The Drosophila melanogaster larval path-length phenotype is an established measure used to study the genetic and environmental contributions to behavioral ...

A Simple Way to Measure Alterations in Reward-seeking Behavior Using Drosophila melanogaster

We describe a protocol for measuring ethanol self-administration in fruit flies (Drosophila melanogaster) as a proxy for changes in reward states. We demonstrate a simple way to tap into the fly reward system, modify experiences related to natural reward, and use voluntary ethanol consumption as a measure for changes in reward states. The approach serves as a relevant tool to study the neurons and genes that play a role in experience-mediated changes of internal state. The protocol is composed of two discrete parts: exposing the flies to rewarding and nonrewarding experiences, and assaying voluntary ethanol consumption as a measure of the motivation to obtain a drug reward. The two parts can be used independently to induce the modulation of experience as an initial step for further downstream assays or as an independent two-choice feeding assay, respectively. The protocol does not require a complicated setup and can therefore be applied in any laboratory with basic fly culture tools.

A Simple Way to Measure Alterations in Reward-seeking Behavior Using Drosophila melanogaster

We describe a protocol for inducing rewarding and nonrewarding experiences in fruit flies (Drosophila melanogaster) using voluntary ethanol consumption as a measure for changes in reward states.

一个简单的方法来衡量在改建悬赏寻求行为使用<em&gt;果蝇</em

We describe a protocol for measuring ethanol self-administration in fruit flies (Drosophila melanogaster) as a proxy for changes in reward states. We ...

Endurance Training Protocol and Longitudinal Performance Assays for Drosophila melanogaster

One of the most pressing problems facing modern medical researchers is the surging levels of obesity, with the consequent increase in associated disorders such as diabetes and cardiovascular disease 1-3. An important topic of research into these associated health problems involves the role of endurance exercise as a beneficial intervention.
Exercise training is an inexpensive, non-invasive intervention with several beneficial results, including reduction in excess body fat 4, increased insulin sensitivity in skeletal muscle 5, increased anti-inflammatory and antioxidative responses 6, and improved contractile capacity in cardiomyocytes 7. Low intensity exercise is known to increase mitochondrial activity and biogenesis in humans 8 and mice, with the transcriptional coactivator PGC1-&alpha; as an important intermediate 9,10. 
Despite the importance of exercise as a tool for combating several important age-related diseases, extensive longitudinal genetic studies have been impeded by the lack of an endurance training protocol for a short-lived genetic model species. The variety of genetic tools available for use with Drosophila, together with its short lifespan and inexpensive maintenance, make it an appealing model for further study of these genetic mechanisms. With this in mind we have developed a novel apparatus, known as the Power Tower, for large scale exercise-training in Drosophila melanogaster 11. The Power Tower utilizes the flies' instinctive negative geotaxis behavior to repetitively induce rapid climbing. Each time the machine lifts, then drops, the platform of flies, the flies are induced to climb. Flies continue to respond as long as the machine is in operation or until they become too fatigued to respond. Thus, the researcher can use this machine to provide simultaneous training to large numbers of age-matched and genetically identical flies. Additionally, we describe associated assays useful to track longitudinal progress of fly cohorts during training.

Endurance Training Protocol and Longitudinal Performance Assays for Drosophila melanogaster

We describe the first endurance training protocol for an important genetic model species, Drosophila melanogaster, and outline several assays to chart improvements in mobility following training.

果蝇的耐力训练协议和纵向性能检测

One of the most pressing problems facing modern medical researchers is the surging levels of obesity, with the consequent increase in associated disorders ...

Fly Climbing Assay: A Method to Test General Locomotion in Drosophila melanogaster

Source: Gevedon, O., et al. <a href="https://www.jove.com/video/59720/in-vivo-forward-genetic-screen-to-identify-novel-neuroprotective">In Vivo Forward Genetic Screen to Identify Novel Neuroprotective Genes in Drosophila melanogaster</a>. J. Vis. Exp. (2019).
This video describes a simple method to quantify fly locomotion and shows the climbing assay performed.&#160; The fly climbing assay is a general test of motor function which relies on a negative geotaxis response.

苍蝇攀爬试验:一种测试黑腹果蝇一般运动的方法

Source: Gevedon, O., et al. In Vivo Forward Genetic Screen to Identify Novel Neuroprotective Genes in Drosophila melanogaster. J. Vis. Exp. (2019).
This ...

JoVE Encyclopedia of Experiments

黑腹果蝇（果蝇）

成虫

Lifespan Protocol for Drosophila melanogaster: Generating Survival Curves to Identify Differences in Fly Longevity

Source: Bovier, T. F., et al. <a href="https://www.jove.com/video/59535/" target="_blank">Methods to Test Endocrine Disruption in Drosophila melanogaster</a>. J. Vis. Exp. (2019).
Scientists use the Drosophila lifespan protocol to measure how different experimental conditions affect flies&#39; longevity and generate survival curves in the process to quantify the experimental outcomes. The sample protocol features the assay being applied to study the effects on longevity from exposure to varying concentrations of endocrine disruptor chemicals, or EDCs, such as 17-&#945;-ethinylestradiol (EE2).

黑腹果蝇的寿命方案:生成生存曲线以确定果蝇寿命的差异

Source: Bovier, T. F., et al. Methods to Test Endocrine Disruption in Drosophila melanogaster. J. Vis. Exp. (2019).
Scientists use the Drosophila lifespan ...

TreadWheel:果蝇轻度诱导运动的间歇训练协议

摘要

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此视频中的章节

含量自发活动，研究昼夜节律和睡眠参数果蝇

使用旋转运动量化系统测量果蝇中的运动水平 (满足要求)

高分辨率视频跟踪移动的，在成年果蝇。

An Affordable and Efficient "Homemade" Platform for Drosophila Behavioral Studies, and an Accompanying Protocol for Larval Mitochondrial Respirometry

一种测定果蝇运动活性的简单技术

觅食路径长度协议

一个简单的方法来衡量在改建悬赏寻求行为使用

果蝇的耐力训练协议和纵向性能检测

苍蝇攀爬试验:一种测试黑腹果蝇一般运动的方法

黑腹果蝇的寿命方案:生成生存曲线以确定果蝇寿命的差异