Temperature is a critical and continuous environmental factor that directly affects biochemical processes within organisms. Animals may habituate to environmental temperature change using a range of mechanisms. To investigate mechanisms of temperature habituation, we exposed Caenorhabditis elegans nematodes to a temperature of 2°C, which is much colder than their normal growing temperature (approximately 10–28°C) and we evaluated their survival rate in recovery from this cold shock. The survival rate after cold shock was different between animals grown at 15°C and 25°C, suggesting that C. elegans exhibits temperature-habituation-linked cold tolerance. Here we show in detail two protocols: a standard cold-tolerance assay after cultivation at constant temperature; and a multistep temperature-shifted cold-tolerance assay.
(T.U. & A.O. contributed equally to this work.)