A Temperature Gradient Assay to Determine Thermal Preferences of Drosophila Larvae.
A Temperature Gradient Assay to Determine Thermal Preferences of Drosophila Larvae.
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DOI：
10.3791/57963
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发表时间：
2018-06-25
期刊：
影响因子：
--
通讯作者：
Montell C
中科院分区：
文献类型：
--
作者：
Liu J;Sokabe T;Montell C
许多动物，包括果蝇，黑腹果蝇，能够分辨出环境温度的细微差异，这使它们能够找到自己喜欢的热景观。为了在定义的线性范围内定义幼虫的温度偏好，我们开发了一种使用温度梯度的分析方法。为了建立单向梯度，两个铝块连接到独立的水浴，每个水浴控制单独块的温度。这两个块设置了渐变的下限和上限。温度梯度是通过将一块涂有琼脂糖的铝板放在两个水控制块上来建立的，这样板就可以跨越它们之间的距离。设置在水块顶部的铝板的两端定义了最低和最高温度，两块之间的区域形成了线性的温度梯度。梯度分析可用于不同年龄的幼虫，并可用于鉴定表现出表型的突变，例如那些影响编码Trp通道和Opsins的基因的突变，这是温度区分所必需的。在这里，我们提出了一种协议来确定果蝇幼虫的首选环境温度使用连续的温度梯度。
Many animals, including the fruit fly, Drosophila melanogaster, are capable of discriminating minute differences in the environmental temperature, which enables them to seek out their preferred thermal landscape. To define the temperature preferences of larvae over a defined linear range, we developed an assay using a temperature gradient. To establish a single-directional gradient, two aluminum blocks are connected to independent water baths, each of which controls the temperature of individual blocks. The two blocks set the lower and upper limits of the gradient. The temperature gradient is established by placing an agarose-coated aluminum plate over the two water-controlled blocks so that the plate spans the distance between them. The ends of the aluminum plate that is set on the top of the water blocks defines the minimum and maximum temperatures, and the regions in-between the two blocks form a linear temperature gradient. The gradient assay can be applied to the larvae of different ages and can be used to identify mutants that exhibit phenotypes, such as those with mutations affecting genes encoding TRP channels and opsins, which are required for temperature discrimination. Here, we present a protocol to determine the preferred environmental temperature of Drosophila larvae using a continuous thermal gradient.
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