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JoVE Encyclopedia of Experiments
Biology
苍蝇攀爬试验:一种测试黑腹果蝇一般运动的方法
苍蝇攀爬试验:一种测试黑腹果蝇一般运动的方法
Encyclopedia of Experiments
Biology
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Encyclopedia of Experiments Biology
Fly Climbing Assay: A Method to Test General Locomotion in Drosophila melanogaster

苍蝇攀爬试验:一种测试黑腹果蝇一般运动的方法

Protocol
13,318 Views
02:33 min
April 30, 2023
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- 首先,获得一定数量的成年果蝇。攀爬测定的测试室由两个用胶带粘在一起的小瓶组成。其中一个样品瓶应标有一条线。

如果没有二氧化碳麻醉(可能会损害攀爬行为),请将一定数量的果蝇转移到测试室。让果蝇恢复预定的时间。将腔室的标记面朝下,用力敲击放置在固体表面上的鼠标垫上的腔室。这将迫使苍蝇到达腔室的底部。攀爬测定利用了果蝇逆重力向上攀爬的自然倾向,这种行为称为负趋向性。

在试验期间,果蝇不应暴露在阳光直射下,因为果蝇可能会表现出趋光性行为或响应光刺激的运动。记录到达或越过线路的苍蝇数量。然后,通过将该数字除以测试的果蝇总数来获得通过率百分比。在重复试验之前让果蝇恢复。在该示例中,我们将观察到用于评估化学诱变后果蝇运动功能的苍蝇攀爬试验。

-首先,从属于映射到第二染色体的 ENU 诱变果蝇突变体集合的品系中收集纯合果蝇。将 10 至 12 天大的果蝇翻转到没有麻醉的测试室中,一次一根线,让它们恢复 5 分钟。

将 5 厘米线标记为正面朝下,在放置在固体表面上的鼠标垫上用力敲击腔室 3 次,以便所有果蝇从底部开始测定。观察苍蝇在 10 秒内的运动。记录在此时间内到达或通过 5 厘米线的苍蝇数量以及测试的苍蝇总数。等待一分钟,然后开始第二次试用。每行至少重复 3 次试验重复。

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