Abstract
Many animals, including the fruit fly, Drosophila melanogaster, are capable of discriminating minute differences inenvironmental 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 tolarvae 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.
Highlights
Thermotaxis is employed by mobile animals to select an environment with the most favorable conditions[1,2,3]
Assays to monitor larval thermotaxis have been instrumental in identifying and clarifying the roles of molecular sensors such as Drosophila Transient Receptor Potential (TRP) channels[4,5,6], rhodopsins[7,8], and ionotropic receptor receptors (IRs)[9], which endow these animals with temperature sensitivities over different temperature ranges
After confirming that there is no adult bodies or food debris floating on the surface of the sucrose solution, pour the top layer containing the larvae through the strainer to trap all the larvae on the mesh screen
Summary
Thermotaxis is employed by mobile animals to select an environment with the most favorable conditions[1,2,3]. Since Petri dishes are round, the sizes of the temperature zones vary and are progressively smaller depending on the distance from the center This setup is not ideal for monitoring the temperature selections of populations of animals. A continuous thermal gradient apparatus that is well-suited to assess the temperature preferences of groups of larvae employs a rectangular arena and is described here. In addition to reporting the favored temperature of larvae, it reveals the preferences of the population of animals over an entire linear range in a single experiment. Due to these advantages, it is an excellent choice for identifying genes required for thermotaxis
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