Name: experimental protocol for using drosophila as an invertebrate model system for toxicity testing in the laboratory
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这份出版物得到了教育部的资助 (PR 奖 #P031C160025 17, 项目标题: 84.031C) 到科罗拉多州立大学-普韦布洛 (CSU-普韦布洛) 社区, 以建立积极的干细胞参与 (C 基地)。我们感谢目前的动物学和 Elsevier, 提供使用前几份论文中发表的代表性成果的权利, 以及朱庇特的编辑们为我们提供了发表本议定书的机会。我们还要感谢 C 基地计划, 布莱恩凯文·范德·艾格 Heuvel 博士 (c 基地和生物学系, csu-普韦布洛), csu-普韦布洛生物系, 托马斯格拉齐亚诺, 伯纳德 Possidente 博士 (生物学系, 斯基德莫尔学院) 和克莱尔博士拉莫斯(科罗拉多州立大学生物系) 给予他们的支持和帮助。

下面的协议是一种实验性的协议, 用于后方物种在果蝇属的污染培养基, 当口服摄取毒素是适当的;其他形式的暴露是可能的使用果蝇模型11,15,16,26。本协议中描述的方法以前是由. 19和彼得森等。23,24,25。
1. 在研究实验室建立果蝇种群
<ol><li>建立一个由环境控制的孵化器 (或小房间), 通过设置恒温器, 光照: 暗循环和湿度, 取决于测试物种的喜好, 为果蝇的种群提供住房。 注意: 首选的环境条件将因该研究所选择的物种的本土生态学而异。例如, 黑腹是原产于撒哈拉以南非洲27并且通常保持在25°c, 12:12 光: 黑暗的周期, 并且大约60% 湿气16,18,19,20 ,21,22,23,24,25,28,29,30. 另一方面,蒙大拿州的范围遍及加拿大和美国中西部, 这是一个更寒冷的地区;因此,蒙大拿通常保持在19–20°c, 有时是24小时的光政权, 以模拟条件在交配季节31。有关不同种类果蝇的地理范围的更详细的描述, 请参见果蝇形态模式网站32。</li>
	<li>
获得一个首选的果蝇品种和/或自交系从一个股票中心 (见资料表), 另一研究实验室应要求, 或收集野生, 基因变量的人口从外地。 注: 以下步骤解释了收集野生的、基因变量的果蝇种群以维持在研究实验室中的方法。这些方法已从马尔可夫和 O ' 格雷迪33和沃纳和 Jaenike34 , 以收集最广泛的物种, 一次, 而不是针对特定物种与一个诱饵来源。<ol>
<li>在冷藏库中冻结半打成熟的香蕉过夜, 在设置诱饵陷阱之前解冻。</li>
		<li>在瓶子的前面切一个 u 形的狭缝, 让苍蝇在饵瓶中被捕获, 而不是逃跑, 准备多部的塑料瓶。用瓶盖盖上塑料瓶, 这样苍蝇就不会通过盖子逃走。</li>
		<li>把解冻香蕉放到瓶子的底部, 这样瓶子的底部就含有大约一英寸的香蕉。把一大块成熟的西红柿放在瓶子里。将酵母浆 (从啤酒酿造过程中残留的酵母) 添加到瓶子底部的香蕉上, 这样香蕉就能浸泡在酵母浆中。</li>
		<li>添加木杆 (在直立垂直位置) 的瓶子, 使苍蝇有一个干净的基板走在远离酵母浆和香蕉。 图 1说明了这些方法的最终产品。</li>
		<li>一夜之间在树上挂饵瓶, 每24小时检查一次口吸出的瓶子, 并单独放置女性在小瓶中, 以创建 iso 女性线。 注意: 可以创建多雌性线, 但是, 只有当每个物种的雌性可以清楚地辨认。另外,果蝇在种类之内占领不同的生态学利基并且将有不同的膳食要求取决于那些适当位置 (Werner 并且 Jaenike34);参见沃纳和 Jaenike34的饮食建议和食物食谱。</li>
		<li>检查解剖显微镜下的成人 F1子代, 以确定收集的果蝇的种类 (见马尔可夫和 O ' 格雷迪33和沃纳和 Jaenike34 , 以协助鉴定各种物种).</li>
	</ol>
</li>
</ol><img alt="Figure 1" class="xfigimg" src="/files/ftp_upload/57450/57450fig1.jpg"> 图 1: 用于收集野外果蝇野生种群的陷阱和诱饵的图案表示.(a)在科罗拉多州的一个当地实地地点设置了飞行陷阱。(B)更密切地了解这一实地地点设置的飞行陷阱。<a href="/files/ftp_upload/57450/57450fig1large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
<ol start="3"><li>保持在环境控制的孵化器或室内恒温, 光照: 暗循环和湿度的 iso 女性或多女性线。要做到这一点, 房子苍蝇在瓶子或瓶子的首选培养基, 并允许妊娠雌性产卵的培养基。监测小瓶的幼虫和蛹的存在。 注:果蝇内蝇属不同的生态龛位, 将有不同的饮食要求和环境非生物偏好取决于这些利基33,34。在埃尔金和米勒28, 谢弗等地可以找到环境偏好和饮食建议 (以及进一步的畜牧指导)。29, 公牛和英勇的30, 马尔可夫和 O ' 格雷迪33, 和沃纳和 Jaenike34。如果使用野生捕获的物种, 可以在孵化器中模拟当地环境条件, 直到能够识别出物种。</li>
	<li>经常将库存转移到新鲜的培养基上, 丢弃旧瓶子, 保持健康的线条, 避免螨感染。 注意: 转移的频率将取决于物种的生命周期。例如, 每2周将果蝇移植到新鲜培养基中。有关在实验室中维护线路的详细信息,请参阅兰德等。16, 埃尔金和米勒28, 谢弗等。29, 公牛和英勇30, 格林斯潘35和科学教育数据库36。</li>
</ol>2. 受污染介质中的后果蝇
注: 如果测试果蝇在实验室中第一次或与新的污染物 (s), 确定致命剂量 (见卡洛斯·卡斯塔涅达等。37和马西等。38的方法) 和 LD50 (见卡洛斯·卡斯塔涅达等.37和埃金斯等。39为方法) 首先。然后, 运行剂量响应曲线, 以确定所需的表型输出的生物相关浓度;看到了吗？19和周等。40为方法。
<ol><li>根据污染物的化学成分, 在所需浓度 (s) 的情况下, 准备受污染介质的库存溶液。 注: 例如, 准备 PbAc 的库存解决方案: 通过将污染物添加到蒸馏水 (dH20) 中, 准备醋酸铅 (PbAc) 介质的库存溶液, 直到用污染物达到所需浓度的介质。例如, 1000 µM PbAc 的库存解决方案, 可以通过将 PbAc 添加到 dH20 中, 直到达到1000µM PbAc。此外, 稀释库存解决方案 (如1000 µM PbAc 解决方案) 到所需浓度 (如500µM PbAc), 并保持这些解决方案作为库存。</li>
	<li>准备介质, 遵循制造商的指导方针作为控制介质。根据制造商的指导方针, 准备其他介质;不过, 为弶镀 ' p20 制备的污染溶液补充剂。 注: 例如, 如果使用瞬时果蝇培养基, 添加大约一茶匙即时培养基到塑料瓶。向培养基中添加大约5–5.5 毫升 dH20。洒几粒活面包酵母, 准备控制培养基。准备实验介质, 补充20 µM 的库存溶液 (如 500 PbAc)。</li>
	<li>
将繁殖存活成熟的雄性和雌性从种群中转移到控制和实验培养基中。 注: 羽化的时间后, 生殖成熟度是不同的果蝇物种41。<ol>
<li>用占主导地位的手轻轻地敲打一小瓶的股票。确保苍蝇自动移动到瓶子的底部。另一方面, 在轻拍瓶子的同时取下瓶子的瓶盖, 在瓶子顶端放一小瓶控制或污染的培养基。把瓶子放在一起, 翻转过来, 轻轻敲击, 这样苍蝇就会自动转移到控制或污染介质的新鲜瓶子里。趁苍蝇还在敲瓶子的时候, 把瓶子盖上。</li>
		<li>重复与更多的瓶子, 确保标准化的苍蝇在每个瓶子。 注:通过单转移或麻醉进行转移的成年人总数将取决于用于避免过度拥挤的小瓶的大小。</li>
		<li>在标准环境条件下孵化成人 (即孵化器), 允许成年人在培养基中交配和产卵 24–96 h。</li>
		<li>在 24–96 h 之后, 在停尸房里丢弃成年人 (一个装满矿物油的瓶子, 用一个紧的漏斗) 留下受精卵 (后来成为实验对象), 以成熟的测试。把瓶子放进孵化器, 让卵发育。</li>
		<li>通过寻找从培养基中产生的幼虫来监测流浪龄幼虫的小瓶。</li>
	</ol>
</li>
</ol>3. 在不同的发展阶段收集实验对象
注: 实验对象可在任何发育阶段收集, 放置在盲编码的15毫升锥形管中, 并进行测试以积累。污染物积累的测试方法将取决于所研究的污染物。例如, PbAc 的积累可以用电感耦合等离子体质谱 (ICP-MS)42进行测试。此外, 实验对象可以收集在任何发展阶段, 以测试各种表型效应的污染物。图 2说明了果蝇生命周期43。图 3说明了暴露的实验协议和收集的不同发展阶段。
<img alt="Figure 2" class="xfigimg" src="/files/ftp_upload/57450/57450fig2.jpg"> 图 2: 对果蝇生命周期的概念性概述 (最常用的果蝇模型系统).果蝇生命周期的阶段为: 1) 蛋, 2) 第一龄幼虫, 3) 二龄幼虫, 4) 第三龄幼虫, 5) 流浪第三龄幼虫, 6) 白眼蛹, 7) 红眼蛹, 8) 新 eclosed 成人, 9) 成熟成人。<a href="/files/ftp_upload/57450/57450fig2large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
<img alt="Figure 3" class="xfigimg" src="/files/ftp_upload/57450/57450fig3.jpg"> 图 3: 在父母 (f0) 和随后世代 (f1和以后) 对被污染的媒介的果蝇的口头暴露方法的概念概述.(A)暴露的世代在发展过程中的口腔接触方法。(B)测试将污染物转移到后代的方法 (F1到所需的世代)。这个数字已经从彼得森等。24<a href="/files/ftp_upload/57450/57450fig3large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
<ol><li>
收集流浪-第三龄幼虫
	<ol>
<li>启动监控瓶时, 灯打开的孵化器, 因为幼虫将出现从培养基和向上移动的瓶子一侧在 h 后, 灯打开在孵化器。在这个 h 中, 用木棍或镊子小心地从瓶子两侧移走流浪的第三龄幼虫。 注: 可供采集的幼虫数量将取决于 "2.3.4" 中产卵的数量。</li>
		<li>要去除幼虫中多余的培养基, 将幼虫放在一个小烧杯中, 用 dh2o 将 dh2O 倒入烧杯中, 将幼虫放在一个微妙的任务雨刷上。使用一个微妙的任务雨刷, 轻轻地去除多余的 dH2O 从幼虫。</li>
		<li>在环境控制的孵化器中维持实验种群。</li>
	</ol>
</li>
	<li>
收集新 eclosed 的成年人
	<ol>
<li>通过观察小瓶两侧的蛹的颜色来监测羽化的瓶子。 注意: 蛹在发育过程中会变暗。发育时间, 特别是预羽化, 取决于所测试的物种。</li>
		<li>当第一个成年人开始 eclose 的时候, 把这些成年人扔进一个含矿物油的太平间里。</li>
		<li>第二天早晨, 当在孵化器中打开灯时, 倾倒并丢弃任何可能在夜间或 morningbefore 灯亮起 eclosed 的未知年龄 (或童贞) 的成年人。</li>
		<li>大约4小时后, 麻醉任何成人出现作为新 eclosed 的成年人与2枪的瓶子。在解剖显微镜下将成人放置在 CO2板上。性成人通过寻找性别梳子的男性和 ovipositors 在女性的前肢。 注: 羽化必须在6小时内收集, 以避免交配, 但其他物种可能有较长的发育时间 (因此, 不需要在这个时间范围内收集)。</li>
		<li>用木棍在 CO2板上分离成人。用木棍轻轻地将成年人转移到与性别相关的群体中, 以此来匹配先前存在的历史。</li>
	</ol>
</li>
	<li>
收集成年后羽化
	<ol>
<li>允许成年人留在培养基匹配前羽化暴露从卵阶段到预期年龄后羽化在一个环境控制的孵化器。</li>
		<li>在测试前, 将成年人单独转移到控制培养基上24小时, 以允许成年人从身体上培养过量的污染介质。</li>
	</ol>
</li>
</ol>4. 后方实验对象测试几代或跨代暴露的影响。
<ol><li>后方父母世代 (ak a P0或 F0代), 转移成人从存货人口到控制和实验性媒介跟随步骤在 "2.1" 到 "2.3" 和 "3.1" 到 "3.3"。</li>
	<li>成人繁殖成熟时 (见 Pitnick等.41), 单独转移 (如2.3.1 所述) 一小瓶雄性到一瓶新的控制或实验培养基。单独转移一小瓶女性到现在包含男性的新鲜的小瓶。允许成年人在培养基中交配产卵24-96 小时, 然后将成年人丢弃在含有矿物油的太平间里, 重新孵化瓶子, 让后代发育。</li>
	<li>根据所需的世代数重复步骤4.1 到4.2。</li>
</ol>

作者没有什么可透露的。

果蝇作为一个强大的模型, 为一系列的生物过程, 由于广泛的保护基因和通路之间的人和人类 13,14。由于同样的原因, 它是一个强大的医学模型,果蝇已经成为一个合适的模型系统来研究人类污染对一系列毒理学端点的影响。几个实验室成功地使用了D.黑腹作为模型系统研究一系列化合物, 包括重金属11,16,18-25,37 ,38,39,40,44,45, 乙醇46, 纳米粒子26,47, 杀虫剂48和溶剂49。尽管最近努力利用果蝇作为毒理学模型, 它作为一个模型系统来回答无数毒理学问题仍处于起步阶段。然而, 鉴于它广泛用作医学相关端点的模型, 以及它在生态学50和进化研究17中的应用, 它作为毒理学模型系统的潜力是巨大的。
在此, 我们提出了在受污染培养基上培育果蝇属中各种物种的方法, 以测试各种毒理学端点。虽然其他形式的暴露是可能的使用果蝇作为模型 (如吸入和皮肤接触), 本议定书的重点是对污染物的口头消费, 这是必要的污染物, 自然会被摄取 (比如通过食物链)。这些方法可以适应多种果蝇种类和污染物的使用。野生的, 基因变量的果蝇种群也可以收集在田间和维持在研究实验室。有许多选择的陷阱和诱饵, 可以使用, 取决于物种的食物偏好;有关字段集合的字段指南, 请参阅马尔可夫和 O ' 格雷迪33和沃纳和 Jaenike34。此外, 这些方法可以改变, 以确定发展暴露在不同的关键发展时期的影响, 并允许长期多代测试污染物暴露。
这些方法的关键步骤包括: (1) 在环境控制条件下维持飞行种群, (2) 避免飞行种群过度拥挤, (3) 根据化学性质稀释试验污染物, (4) 选择测试污染物的生物相关浓度。在环境调节的孵化器 (或一间小房间) 内维持库存, 可确保在不同条件下的额外变化不会混淆结果。此外, 以往的行为季节性变化已被发现51和几个果蝇物种进入滞育在冬季52。第二, 幼虫过度拥挤对发育30、成人体型30和长寿53有长远影响。此外, 污染物的稀释是确保果蝇在生物上可以积累污染物的重要步骤。例如, PbAc 溶解在 dH2O23,24,25, 而其他化学品可能需要溶解在盐水或乙醇。选择与生物相关的污染物浓度会影响结果的方向;例如 , 低剂量的 PbAc 增加了与雄配的平均数雌性 ( 20 分钟内 ) , 而较高的剂量显示雌配19的平均数量显著下降。为了确定测试污染物的生物相关浓度, 读者应考虑运行初步研究, 以确定致命剂量和 LD50, 以确定适当剂量, 以执行剂量反应曲线。通过执行剂量响应曲线来测试某一特定端点上的浓度范围, 读者可以确定对个人或人群来说, 对其进行进一步测试的剂量是 "有益的" 或 "危险的"。
该协议提供了一个途径来确定: 1) 组织的多重生物水平对健康和毒理学终点的相互作用;2) 发展和紧急因素的作用;3) 生态重要端点;4) 医学上-重要端点;5) 多重压力源如何相互作用产生结果;和 6) 长期暴露的影响, 超越世代。为了说明本议定书的有效性, 提供了证据表明, 在整个发展过程中暴露的个人积累了 Pb (表 1)23,24。此外, 具有代表性的结果表明, 该协议可用于测试暴露在生态重要端点 (例如, 发育铅暴露对交配选择的影响24) 的影响。此外, 其他人已经测试了污染物对组织的多重生物水平的影响 (包括生理18,21, 遗传20,22和表型-水平19,23,24,25), 医学上-重要端点 18,20,21,22,23, 和长期多代效应23,24 ,25,54。此外, 初步数据表明, 发育铅暴露诱导跨代遗传效应的生育力在54。这项议定书的一个重要限制是, 使用这项议定书与果蝇处于萌芽阶段。因此, 有有限的出版物18,19,20,21,22,23,24,25解决《议定书》对补充毒理学问题的潜力, 例如发展和紧急因素的作用、额外的生态重要终点、多重压力源以及暴露的演变影响。
使用此协议, 读者可以测试自然摄取的污染物, 使用生物相关的方法。连续液体喂养, 由苏亚雷斯和al开发。55是一种替代的口服方法, 特别是对农药的接触。然而, 连续液体喂养适合成人摄取液体污染物, 不适用于个人可能暴露前羽化的污染物。这一点尤其重要, 因为在开发中可能存在暴露的关键时期。先前的研究显示, 铅暴露的关键时期为23。因此,果蝇应该在整个开发过程中暴露出来, 以避免果蝇在测试之前有可能积极消除污染物, 直到确定关键时期。
总之, 我们提供了一种将果蝇口头暴露于污染物的协议。使用这个协议和模型系统, 毒理学家可以转向伦理和非侵入性的方法, 以动物测试, 同时纳入一个更全面的方法, 以了解污染物的影响8。

每年, 化工行业引进约1000种新化学品1、2;然而, 在2、3分销之前, 仅有很小一部分化学物质对环境的影响进行了测试。虽然大规模的灾难是罕见的, 致死和长期接触大量的污染物在人类和野生动物4,5普遍存在。生态和环境毒理学的历史焦点是测试致死性、单一化学暴露、急性暴露和暴露的生理效应, 作为衡量污染物对生存6的影响的一种手段,7,8,9,10. 虽然对动物试验采取了伦理和非侵入性的做法, 但目前的做法受到限制, 因为发展、紧急性质和外部因素 (如人口水平和生态系统级相互作用) 在斡旋生态学重要端点8。因此, 有必要在不牺牲野生动物和/或脊椎动物的实验室中纳入更全面的方法。
无脊椎动物模型系统, 如果蝇, 是一个有吸引力的替代方案, 以解决对毒性测试的更全面的方法的需要。在一个世纪前的11, 它最初被发展为人类相关基因研究的无脊椎动物模型系统。由于以下几个原因, 黑腹现在已被广泛用作脊椎动物模型的替代品: 1) 保护的基因和通路之间的黑腹和人类;2) 与脊椎动物模型相比短世代时间;3) 维护成本低廉;4) 易于生成大样本尺寸;和 5) 过多的表型和生态相关的端点可用于测试11,12,13,14,15,16,17.
几个实验室11,15,16,17,18,19, 20,21,22,23,24,25现在使用D.黑腹作为一种脊椎动物模型的毒性试验, 以了解污染对人类的影响。当地野生果蝇可以被利用, 以及作为野生动物 (和人类) 的毒性模型, 以回答生态, 行为上和进化相关的问题, 在多个生物层次的组织。利用果蝇属中的物种作为模型, 几个可测量的端点是可能的11,15,16,18,19,20 ,21,22,23,24,25。另外, 使用果蝇模型, 毒理学家可以: 1) 道德上的链接效应在组织的多生物水平;2) 纳入紧急因素和发展的作用;3) 研究生态学上重要端点 (除医学上重要端点);4) 同时测试多个压力源;5) 和测试的长期多代 (如进化和跨代) 的影响的压力。因此, 利用果蝇作为模型系统, 可以采取多种方法, 不限于研究机械方法与自交系的近亲在实验室 。
本文介绍了果蝇的饲养和采集方法, 以回答各种毒理学问题。更具体地说, 我们描述的方法 1) 饲养果蝇的中掺有一个或多个污染物;2) 在整个发育过程中收集果蝇(如流浪的第三龄幼虫、蛹病例、新 eclosed 成年人和成年成年人);3) 在受污染的培养基中饲养果蝇, 以测试代际和跨代的传播, 以及长期毒物暴露对进化的影响。使用此协议, 以前的作者18,19,20,21,22,23,24,25报告了发育铅 (Pb2 +) 暴露的不同生理、遗传和行为效应。这项议定书使毒理学家能够使用一种更整体的毒理学方法, 这对于了解污染物是人类和野生动物在日益污染的环境中的危险因素是必不可少的。

<table><tbody><tr><td>Carolina Biological Instant Drosophila Medium Formula 4-24</td><td>Carolina Biological</td><td>173204</td><td></td></tr><tr><td>Drosophila vials, Narrow (PS), Polystyrene, Superbulk, 1000 vials/unit</td><td>Genessee Scientific</td><td>32-116SB</td><td>Used to store flies</td></tr><tr><td>Flugs Closures for vials and bottles, Narrow plastic vials</td><td>Genessee Scientific</td><td>49-102</td><td>Used to store flies</td></tr><tr><td>Cardboard trays, trays only, narrow</td><td>Genessee Scientific</td><td>32-124</td><td>Used to organize populations of flies</td></tr><tr><td>Cardboard trays, dividers only, narrow</td><td>Genessee Scientific</td><td>32-126</td><td>Used to organize populations of flies</td></tr><tr><td>Thermo Scientific&amp;nbsp;Nalgene Square Wide-Mouth HDPE Bottles with Closure</td><td>Fischer Scientific</td><td>03-312D</td><td>Useful for storage of contaminants</td></tr><tr><td>Thermo Scientific&amp;nbsp;Nalgene Color-Coded LDPE Wash Bottles</td><td>Fischer Scientific</td><td>03-409-17C</td><td>Useful for storage of contaminants</td></tr><tr><td>Eppendorf&amp;nbsp;Repeater M4 Manual Handheld Pipette Dispenser</td><td>Fischer Scientific</td><td>14-287-150</td><td>Used to prepare medium</td></tr><tr><td>Combitips Advanced Pipetter Tips - Standard, Eppendorf Quality Tips</td><td>Fischer Scientific</td><td>13-683-708</td><td>Used to prepare medium</td></tr><tr><td>Flypad, Standard Size (8.1 X 11.6cm)</td><td>Genessee Scientific</td><td>59-114</td><td>Used to anesthetize flies</td></tr><tr><td>Flystuff foot valve</td><td>Genessee Scientific</td><td>59-121</td><td>Used to anesthetize flies</td></tr><tr><td>Tubing, green (1 continguous foot/unit)</td><td>Genessee Scientific</td><td>59-124G</td><td>Used to anesthetize flies</td></tr><tr><td>Mineral Oil, Light, White, High Purity Grade, 500 mL HDPE Bottle</td><td>VWR</td><td>97064-130</td><td>Used to make a morgue</td></tr><tr><td>Glass Erlenmeyer Flask Set - 3 Sizes - 50, 150 and 250ml, Karter Scientific 214U2</td><td>Walmart</td><td>Not applicable</td><td>Used to make a morgue</td></tr><tr><td>BGSET5 Glass Beaker Set Of 5</td><td>Walmart</td><td></td><td></td></tr><tr><td>Inbred or wildtype line of &lt;em&gt;Drosophila&lt;/em&gt;</td><td>Bloomington &lt;em&gt;Drosophila&lt;/em&gt; Stock Center at Indiana University</td><td></td><td>https://bdsc.indiana.edu</td></tr><tr><td>Wild popultions of &lt;em&gt;Drosophila&lt;/em&gt;</td><td>UC San Diego &lt;em&gt;Drosophila&lt;/em&gt; Stock Center</td><td></td><td>https://stockcenter.ucsd.edu/info/welcome.php</td></tr></tbody></table>

experimental protocol for using drosophila as an invertebrate model system for toxicity testing in the laboratory

紧急性质和外部因素 (特别是人口水平和生态系统层面的相互作用) 在生态重要的端点的调解中起着重要作用, 尽管它们在毒理学研究中很少被考虑。作为毒物的行为、神经和遗传影响的毒理学模型, 腹腹已经成为了一个典型。更重要的是,果蝇属中的物种可以作为一种综合框架方法的模型系统, 在毒物学研究中纳入紧急性质并回答生态相关问题。本文的目的是提供一个协议, 以揭露果蝇属物种的污染物, 作为一个模型系统的一系列表型产出和生态相关的问题。更具体地说, 本议定书可用于 1) 将组织的多个生物层次联系起来, 了解毒物对个人和人口水平的影响;2) 测试毒物在发育暴露的不同阶段的影响;3) 检验污染物的几代和演变影响;4) 同时测试多种污染物和压力源。

通过将果蝇口头暴露在整个开发过程中, 可以通过将果蝇暴露在不同的生物组织水平来测试各种毒理学问题。本节介绍在以前发表的论文23、24中使用本协议获得的代表性结果。特别是, 本议定书以前用于评估在同一世代的暴露和跨第一代后代23中铅 (Pb) 的积累、消除和封存;并研究了积累对交配选择24的意义。
表 1和图 4显示了使用此协议获得的具有代表性的结果, 用于确定 f0和 f1代中 Pb 的累积和消除。
表 1显示了在多个发育阶段 (徘徊的第三龄幼虫, 在不同的剂量: 0、10、40、50、75、100、250和500µM PbAc) 的样品中铅累积的代表性结果蛹病例, 新 eclosed 的成年人, 成熟的女性和男性) 在彼得森等。在不同发育阶段收集了23份样品, 冷冻-20 摄氏度, 用硝酸和过氧化氢处理, 用 ICP-MS23,42测定 Pb。
<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always"><tbody>
<tr>
<td>剂量 (µM PbAc)</td>
			<td style="text-align: center;">幼虫</td>
			<td style="text-align: center;">蛹案例</td>
			<td style="text-align: center;">新 Eclosed 成人</td>
			<td style="text-align: center;">成熟的成年女性</td>
			<td style="text-align: center;">成熟成年男性</td>
		</tr>
<tr>
<td>0</td>
			<td style="text-align: center;">0.009 @ 0.001</td>
			<td style="text-align: center;">0.099 @ 0.02</td>
			<td style="text-align: center;">0.04 @ 0.005</td>
			<td style="text-align: center;">0.069 @ 0.031</td>
			<td style="text-align: center;">0.0015 @ 0.003</td>
		</tr>
<tr>
<td>10</td>
			<td style="text-align: center;">0.42 @ 0.025</td>
			<td style="text-align: center;">0.46 @ 0.03</td>
			<td style="text-align: center;">0.12 @ 0.009</td>
			<td style="text-align: center;">0.12 @ 0.012</td>
			<td style="text-align: center;">0.056 @ 0.008</td>
		</tr>
<tr>
<td>40</td>
			<td style="text-align: center;">7.5 @ 0.67</td>
			<td style="text-align: center;"></td>
			<td style="text-align: center;"></td>
			<td style="text-align: center;"></td>
			<td style="text-align: center;"></td>
		</tr>
<tr>
<td>50</td>
			<td style="text-align: center;">5.2 @ 0.71</td>
			<td style="text-align: center;">2.77 @ 0.30</td>
			<td style="text-align: center;">1.11 @ 0.14</td>
			<td style="text-align: center;"></td>
			<td style="text-align: center;"></td>
		</tr>
<tr>
<td>75</td>
			<td style="text-align: center;">14 @ 1.06</td>
			<td style="text-align: center;"></td>
			<td style="text-align: center;"></td>
			<td style="text-align: center;"></td>
			<td style="text-align: center;"></td>
		</tr>
<tr>
<td>100</td>
			<td style="text-align: center;">12 @ 0.95</td>
			<td style="text-align: center;">4.7 @ 0.38</td>
			<td style="text-align: center;">3.36 @ 0.58</td>
			<td style="text-align: center;">1.20 @ 0.63</td>
			<td style="text-align: center;">1.24 @ 0.46</td>
		</tr>
<tr>
<td>250</td>
			<td style="text-align: center;">89 @ 8.49</td>
			<td style="text-align: center;"></td>
			<td style="text-align: center;"></td>
			<td style="text-align: center;">25.06 @ 4.72</td>
			<td style="text-align: center;">5.46 @ 0.75</td>
		</tr>
<tr>
<td>500</td>
			<td style="text-align: center;">271 @ 41.01</td>
			<td style="text-align: center;">334.30 @ 39.43</td>
			<td style="text-align: center;">8.44 @ 0.84</td>
			<td style="text-align: center;">46.50 @ 14.72</td>
			<td style="text-align: center;">14.43 @ 1.83</td>
		</tr>
</tbody></table>表 1: 平均铅负荷 (ng/个人) 测试 在从卵期到试验阶段, 口服铅后, 在发育过程中发生黑腹。手段 (ng/飞行) 的标准误差显示 (n = 8 幼虫, n = 3 控制饲养的成人, n = 3 Pb 饲养的成人)。野生型黑腹黑腹养殖控制或含铅培养基 (0, 10, 40, 50, 75, 100, 250 或500µM PbAc) 从卵阶段到不同的发展阶段。采用 ICP-42 -质谱法收集和试验了铅积累的样品. 这张桌子已经从彼得森和al修改过了。23
在图 4中, 父母世代 (f0) 接触到 Pb 从卵子阶段到成年, 交配在控制培养基中, 第一代后代 (f1) 在控制培养基中饲养直到成年24。检测铅积累和清除的方法类似于彼得森等。23. 这项实验的结果表明, 父母的接触不会传给第一代成年后代24。因此, 使用此协议, 可以测试不同进化尺度下的自适应响应, 以及 F0暴露的跨代效应。在彼得森和al也发现了类似的结果。23
<img alt="Figure 4" class="xfigimg" src="/files/ftp_upload/57450/57450fig4.jpg"> 图 4:Pb 积累在 ( A) 父母 (f0) 和 (B) 未曝光的后代 (f1).在 ( a)和(b)中的条形显示平均值 (ng/成人) ±SEM. 在 (a) 和 (b) 栏上方显示的样本大小。= p < 0.001。(A) 0名成人经口头接触250µM PbAc, 使用本议定书从卵子阶段到 5 d 岁后羽化, 并收集6天后羽化 (24 小时净化) 后, 使用 ICP-MS.42(B) 进行 Pb 堆积试验。0例成人在控制培养基治疗中交配。从卵期到成年 (使用本协议) 在控制培养基中饲养未暴露的 F1子代, 并采用 ICP-MS 进行铅积累试验。在 (B): "cf + cm"= f1成人与父母在控制媒介饲养, "CF + PbM" = f1个成人与父亲在含铅的媒介饲养, "PbF + CM" = f1个成人与母亲在含铅的媒介饲养, "PbF + PbM" = f1成人与父母在含铅培养基饲养。这个数字已经从彼得森等。24<a href="/files/ftp_upload/57450/57450fig4large.jpg" target="_blank">请单击此处查看此图的较大版本.</a> 
表 1和图 4所示的结果表明,果蝇在不同剂量、发育阶段和使用本议定书的进化尺度上都能很容易地积累 Pb。因此, 这表明协议的有效性, 揭露D.
在图 5中, 这里描述的协议被彼得森等使用。24测试发育铅暴露对交配偏好的影响。实验对象从卵期到成年, 在控制或含铅培养基从卵阶段到成年期, 并在24小时净化后测试交配首选。彼得森等。24发现铅暴露的女性优先交配与铅暴露的男性, 当考虑到一个控制或铅暴露的男性的选择。这些结果是执行《议定书》以检查表单输出的一个典型例子。
<img alt="Figure 5" class="xfigimg" src="/files/ftp_upload/57450/57450fig5.jpg"> 图 5:男性和女性的交配偏好在250µM PbAc 从卵子阶段到成年.酒吧在 (A), (B) 和 (C) 显示平均百分比 (%) 交配成功 (在60分钟) 的电子显微镜. ** = p < 0.001. * = p < 0.05。实验对象 (A), (B) 和 (C) 被暴露在控制或含铅培养基 (250 µM PbAc) 从卵子阶段到成熟成年和测试的差异, 配偶选择。(A)女性偏爱控制或 Pb 饲养的雄性 (即两种选择试验)。样品大小是: N = 126 控制饲养的女性和 137 Pb 饲养的女性。(B)男性偏爱控制和铅饲养的雌性 (即两种选择试验)。样品大小是: N = 59 控制饲养的男性和 N = 64 Pb 饲养的男性。(C)男性和女性的交配偏好, 当单独配对一个伴侣的任何接触 (即无选择测试)。在 (C): "CF + CM" = 一个控制饲养的女性配对与一个控制饲养的男性 (N = 85 对), "CF + PbM" = 一个控制饲养的女性配对与一个铅饲养男性 (n = 79 对), "PbF + CM" = 一 Pb 饲养的女性配对与一个控制饲养的男性 (N = 91 对), "PbF +PbM "= 一 pb 饲养的女性 + 一 pb 饲养的男性 (N = 98 对)。这个数字已经从彼得森等。24.<a href="/files/ftp_upload/57450/57450fig5large.jpg" target="_blank">请点击这里查看这个数字的更大版本.</a>

Watch this Scientific Journal Video about Experimental Protocol for Using Drosophila As an Invertebrate Model System for Toxicity Testing in the Laboratory at JoVE.com

Experimental Protocol for Using Drosophila As an Invertebrate Model System for Toxicity Testing in the Laboratory

在本文中, 我们提供了一个详细的协议, 以暴露在果蝇属物种的污染物, 目的是研究暴露对一系列表型产出的影响, 在不同的发展阶段和超过一个世代。

利用果蝇作为无脊椎动物毒性试验模型系统的实验协议

Emergent properties and external factors (population-level and ecosystem-level interactions, in particular) play important roles in mediating ...

experimental-protocol-for-using-drosophila-as-an-invertebrate-model

Universidad San Sebastian

Research

JoVE Journal

Biology

Experimental Protocol for Using Drosophila As an Invertebrate Model System for Toxicity Testing in the Laboratory

12.6K 次观看.  Colorado State University-Pueblo. 在本文中, 我们提供了一个详细的协议, 以暴露在果蝇属物种的污染物, 目的是研究暴露对一系列表型产出的影响, 在不同的发展阶段和超过一个世代。

视频: Experimental Protocol for Using Drosophila As an Invertebrate Model System for Toxicity Testing in the Laboratory

Assaying Locomotor, Learning, and Memory Deficits in Drosophila Models of Neurodegeneration

Advances in genetic methods have enabled the study of genes involved in human neurodegenerative diseases using Drosophila as a model system1. Most of these diseases, including Alzheimer's, Parkinson's and Huntington's disease are characterized by age-dependent deterioration in learning and memory functions and movement coordination2. Here we use behavioral assays, including the negative geotaxis assay3 and the aversive phototaxic suppression assay (APS assay)4,5, to show that some of the behavior characteristics associated with human neurodegeneration can be recapitulated in flies. In the negative geotaxis assay, the natural tendency of flies to move against gravity when agitated is utilized to study genes or conditions that may hinder locomotor capacities. In the APS assay, the learning and memory functions are tested in positively-phototactic flies trained to associate light with aversive bitter taste and hence avoid this otherwise natural tendency to move toward light. Testing these trained flies 6 hours post-training is used to assess memory functions. Using these assays, the contribution of any genetic or environmental factors toward developing neurodegeneration can be easily studied in flies.

Assaying Locomotor, Learning, and Memory Deficits in Drosophila Models of Neurodegeneration

Behavior assays for measuring locomotor functions, learning, and memory abilities in Drosophila.

含量的运动能力，学习和记忆的赤字果蝇</em神经退行性疾病>模型

Advances in genetic methods have enabled the study of genes involved in human neurodegenerative diseases using Drosophila as a model system1. Most of ...

科研

神经科学

Methods to Assay Drosophila Behavior

Drosophila melanogaster, the fruit fly, has been used to study molecular mechanisms of a wide range of human diseases such as cancer, cardiovascular disease and various neurological diseases1. We have optimized simple and robust behavioral assays for determining larval locomotion, adult climbing ability (RING assay), and courtship behaviors of Drosophila. These behavioral assays are widely applicable for studying the role of genetic and environmental factors on fly behavior. Larval crawling ability can be reliably used for determining early stage changes in the crawling abilities of Drosophila larvae and also for examining effect of drugs or human disease genes (in transgenic flies) on their locomotion. The larval crawling assay becomes more applicable if expression or abolition of a gene causes lethality in pupal or adult stages, as these flies do not survive to adulthood where they otherwise could be assessed. This basic assay can also be used in conjunction with bright light or stress to examine additional behavioral responses in Drosophila larvae. Courtship behavior has been widely used to investigate genetic basis of sexual behavior, and can also be used to examine activity and coordination, as well as learning and memory. Drosophila courtship behavior involves the exchange of various sensory stimuli including visual, auditory, and chemosensory signals between males and females that lead to a complex series of well characterized motor behaviors culminating in successful copulation. Traditional adult climbing assays (negative geotaxis) are tedious, labor intensive, and time consuming, with significant variation between different trials2-4. The rapid iterative negative geotaxis (RING) assay5 has many advantages over more widely employed protocols, providing a reproducible, sensitive, and high throughput approach to quantify adult locomotor and negative geotaxis behaviors. In the RING assay, several genotypes or drug treatments can be tested simultaneously using large number of animals, with the high-throughput approach making it more amenable for screening experiments.

Methods to Assay Drosophila Behavior

Drosophila melanogaster is a genetically and behaviorally tractable model system that has been used to understand the molecular and cellular basis of many important biological processes for over a century 1. Drosophila has been well exploited to gain insights into the genetic basis of fly behavior.

对检测方法果蝇行为

Drosophila melanogaster, the fruit fly, has been used to study molecular mechanisms of a wide range of human diseases such as cancer, cardiovascular ...

Methods to Test Endocrine Disruption in Drosophila melanogaster

In recent years there has been growing evidence that all organisms and the environment are exposed to hormone-like chemicals, known as endocrine disruptor chemicals (EDCs). These chemicals may alter the normal balance of endocrine systems and lead to adverse effects, as well as an increasing number of hormonal disorders in the human population or disturbed growth and reduced reproduction in the wildlife species. For some EDCs, there are documented health effects and restrictions on their use. However, for most of them, there is still no scientific evidence in this sense. In order to verify potential endocrine effects of a chemical in the full organism, we need to test it in appropriate model systems, as well as in the fruit fly, Drosophila melanogaster. Here we report detailed in vivo protocols to study endocrine disruption in Drosophila, addressing EDC effects on the fecundity/fertility, developmental timing, and lifespan of the fly. In the last few years, we used these Drosophila life traits to investigate the effects of exposure to 17-&#945;-ethinylestradiol (EE2), bisphenol A (BPA), and bisphenol AF (BPA F). Altogether, these assays covered all Drosophila life stages and made it possible to evaluate endocrine disruption in all hormone-mediated processes. Fecundity/fertility and developmental timing assays were useful to measure the EDC impact on the fly reproductive performance and on developmental stages, respectively. Finally, the lifespan assay involved chronic EDC exposures to adults and measured their survivorship. However, these life traits can also be influenced by several experimental factors that had to be carefully controlled. So, in this work, we suggest a series of procedures we have optimized for the right outcome of these assays. These methods allow scientists to establish endocrine disruption for any EDC or for a mixture of different EDCs in Drosophila, although to identify the endocrine mechanism responsible for the effect, further essays could be needed.

Methods to Test Endocrine Disruption in Drosophila melanogaster

Endocrine disruptor chemicals (EDCs) represent a serious problem for organisms and for natural environments. Drosophila melanogaster represents an ideal model to study EDC effects in vivo. Here, we present methods to investigate endocrine disruption in Drosophila, addressing EDC effects on fecundity, fertility, developmental timing, and lifespan of the fly.

检测果蝇黑色素的内分泌干扰的方法

In recent years there has been growing evidence that all organisms and the environment are exposed to hormone-like chemicals, known as endocrine disruptor ...

发育生物学

Taste Preference Assay for Adult Drosophila

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

Taste Preference Assay for Adult Drosophila

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

口味偏好测定成人<em&gt;果蝇</em

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

Toxicity Study of Zinc Oxide Nanoparticles in Cell Culture and in Drosophila melanogaster

Zinc oxide nanoparticles (ZnO NPs) have a wide range of applications, but the number of reports on ZnO NP-associated toxicity has grown rapidly in recent years. However, studies that elucidate the underlying mechanisms for ZnO NP-induced toxicity are scanty. We determined the toxicity profiles of ZnO NPs using both in vitro and in vivo experimental models. A significant decrease in cell viability was observed in ZnO NP-exposed MRC5 lung fibroblasts, showing that ZnO NPs exert cytotoxic effects. Similarly, interestingly, gut exposed to ZnO NPs exhibited a dramatic increase in reactive oxygen species levels (ROS) in the fruit fly Drosophila. More in-depth studies are required to establish a risk assessment for the increased usage of ZnO NPs by consumers.

Toxicity Study of Zinc Oxide Nanoparticles in Cell Culture and in Drosophila melanogaster

We describe a detailed protocol for evaluating the toxicological profiles of zinc oxide nanoparticles (ZnO NPs) in particular, the type of cell death in human MRC5 lung fibroblasts and ROS formation in the fruit fly Drosophila.

细胞培养和果蝇黑色素的氧化锌纳米颗粒的毒性研究

Zinc oxide nanoparticles (ZnO NPs) have a wide range of applications, but the number of reports on ZnO NP-associated toxicity has grown rapidly in recent ...

免疫与感染

Electrophysiological Recording of The Central Nervous System Activity of Third-Instar Drosophila Melanogaster

The majority of the currently available insecticides target the nervous system and genetic mutations of invertebrate neural proteins oftentimes yield deleterious consequences, yet the current methods for recording nervous system activity of an individual animal is costly and laborious. This suction electrode preparation of the third-instar larval central nervous system of Drosophila melanogaster, is a tractable system for testing the physiological effects of neuroactive agents, determining the physiological role of various neural pathways to CNS activity, as well as the influence of genetic mutations to neural function. This ex vivo preparation requires only moderate dissecting skill and electrophysiological expertise to generate reproducible recordings of insect neuronal activity. A wide variety of chemical modulators, including peptides, can then be applied directly to the nervous system in solution with the physiological saline to measure the influence on the CNS activity. Further, genetic technologies, such as the GAL4/UAS system, can be applied independently or in tandem with pharmacological agents to determine the role of specific ion channels, transporters, or receptors to arthropod CNS function. In this context, the assays described herein are of significant interest to insecticide toxicologists, insect physiologists, and developmental biologists for which D. melanogaster is an established model organism. The goal of this protocol is to describe an electrophysiological method to enable the measurement of electrogenesis of the central nervous system in the model insect, Drosophila melanogaster, which is useful for testing a diversity of scientific hypotheses.

Electrophysiological Recording of The Central Nervous System Activity of Third-Instar  Drosophila Melanogaster

This protocol describes a method to record the descending electrical activity of the Drosophila melanogaster central nervous system to enable the cost-efficient and convenient testing of pharmacological agents, genetic mutations of neural proteins, and/or the role of unexplored physiological pathways.

第三星果蝇中枢神经系统活动的电生理记录

The majority of the currently available insecticides target the nervous system and genetic mutations of invertebrate neural proteins oftentimes yield ...

A Drosophila In Vivo Injury Model for Studying Neuroregeneration in the Peripheral and Central Nervous System

The regrowth capacity of damaged neurons governs neuroregeneration and functional recovery after nervous system trauma. Over the past few decades, various intrinsic and extrinsic inhibitory factors involved in the restriction of axon regeneration have been identified. However, simply removing these inhibitory cues is insufficient for successful regeneration, indicating the existence of additional regulatory machinery. Drosophila melanogaster, the fruit fly, shares evolutionarily conserved genes and signaling pathways with vertebrates, including humans. Combining the powerful genetic toolbox of flies with two-photon laser axotomy/dendriotomy, we describe here the Drosophila sensory neuron &#8211; dendritic arborization (da) neuron injury model as a platform for systematically screening for novel regeneration regulators. Briefly, this paradigm includes a) the preparation of larvae, b) lesion induction to dendrite(s) or axon(s) using a two-photon laser, c) live confocal imaging post-injury and d) data analysis. Our model enables highly reproducible injury of single labeled neurons, axons, and dendrites of well-defined neuronal subtypes, in both the peripheral and central nervous system.

A Drosophila In Vivo Injury Model for Studying Neuroregeneration in the Peripheral and Central Nervous System

Here, we present a protocol using the Drosophila sensory neuron - dendritic arborization (da) neuron injury model, which combines in vivo live imaging, two-photon laser axotomy/dendriotomy, and the powerful fly genetic toolbox, as a platform for screening potential promoters and inhibitors of neuroregeneration.

一种研究外周和中枢神经系统 Neuroregeneration 的果蝇体内损伤模型

The regrowth capacity of damaged neurons governs neuroregeneration and functional recovery after nervous system trauma. Over the past few decades, various ...

医学

Topical Application Bioassay to Quantify Insecticide Toxicity for Mosquitoes and Fruit Flies

The continued use of insecticides for public health and agriculture has led to widespread insecticide resistance and hampering of control methods. Insecticide resistance surveillance of mosquito populations is typically done through Centers for Disease Control and Prevention (CDC) bottle bioassays or World Health Organization (WHO) tube tests. However, these methods can result in a high degree of variability in mortality data due to variable insecticide contact with&#160;the insect, the relatively small numbers of organisms tested, extensive variation in mass between populations, and constantly changing environmental conditions, leading to variable outcomes. This paper presents the topical application bioassay, adapted as a high-throughput phenotypic bioassay for both mosquitoes and fruit flies, to test large numbers of insects along&#160;a range of insecticide concentrations.
This assay 1) ensures consistent treatment and insecticide contact with every organism, 2) produces highly specific dose-response curves that account for differences in average mass between strains and sexes (which is particularly important for field-collected organisms), and 3) allows for the calculation of statistically rigorous median lethal doses (LD50), which are necessary for resistance ratio comparisons-an alternative surveillance approach from diagnostic dose mortality, which is also used for larvicide resistance surveillance. This assay will be a complementary tool for accurately phenotyping mosquito populations and, as illustrated using fruit flies, is easily adaptable for use with other insects. We argue that this assay will help fill the gap between genotypic and phenotypic insecticide resistance in multiple insect species.

We describe the methodology and importance of the topical application bioassay to measure insecticide susceptibility in mosquitoes and fruit flies. The presented assay is high-throughput, utilizes insect mass-thus allowing for calculating a mass-relativized lethal dose instead of concentration-and likely has lower variability than other similar methods.

局部应用生物测定以量化蚊子和果蝇的杀虫剂毒性

The continued use of insecticides for public health and agriculture has led to widespread insecticide resistance and hampering of control methods. ...

生物学

Assessment of Chemical Toxicity in Adult Drosophila Melanogaster

Human industries generate hundreds of thousands of chemicals, many of which have not been adequately studied for environmental safety or effects on human health. This deficit of chemical safety information is exacerbated by current testing methods in mammals that are expensive, labor-intensive, and time-consuming. Recently, scientists and regulators have been working to develop new approach methodologies (NAMs) for chemical safety testing that are cheaper, more rapid, and reduce animal suffering. One of the key NAMs to emerge is the use of invertebrate organisms as replacements for mammalian models to elucidate conserved chemical modes of action across distantly related species, including humans. To advance these efforts, here, we describe a method that uses the fruit fly, Drosophila melanogaster,&#160;to assess chemical safety. The protocol describes a simple, rapid, and inexpensive procedure to measure the viability and feeding behavior of exposed adult flies. In addition, the protocol can be easily adapted to generate samples for genomic and metabolomic approaches. Overall, the protocol represents an important step forward in establishing Drosophila as a standard model for use in precision toxicology.

Assessment of Chemical Toxicity in Adult Drosophila Melanogaster

This protocol describes an efficient and inexpensive method that uses liquid media to assess the effects of chemical toxicants on the viability of adult Drosophila melanogaster.

成虫黑腹果蝇化学毒性评估

Human industries generate hundreds of thousands of chemicals, many of which have not been adequately studied for environmental safety or effects on human ...

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.bdigitaluss.remotexs.co/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 ...

利用果蝇作为无脊椎动物毒性试验模型系统的实验协议

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含量的运动能力，学习和记忆的赤字果蝇模型