Cold tolerance assay for studying cultivation-temperature-dependent cold habituation in C. elegans

Abstract

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.

Endoribonuclease ENDU-2 regulates multiple traits including cold tolerance via cell autonomous and nonautonomous controls in Caenorhabditis elegans

“…Cold Tolerance Assay. Cold tolerance was assayed as described in previous studies (4,18). We used uncrowded and well-fed young adult worms as they prepared to lay eggs.…”

The mechanoreceptor DEG‐1 regulates cold tolerance in Caenorhabditis elegans

“…The cold tolerance assay was performed in accordance with previous reports [5,7,8,44]. For this assay, we placed well-fed adult worms onto nematode growth medium (NGM) with 2% (w/v) agar, and then seeded the medium with Escherichia coli OP50 once the worms began laying eggs.…”

Sperm Affects Head Sensory Neuron in Temperature Tolerance of Caenorhabditis elegans

“…Flesh adult animal(s) were placed on a nematode growth medium (NGM), on which Escherichia coli OP50 was seeded; the animals were transferred to the outside of the plate after 8-12 hr. The plates containing their progenies at the fresh adult stage were transferred to 2 C in a refrigerated cabinet for the designated time (Ohta et al, 2014;Ujisawa et al, 2014). See also the Supplemental Experimental Procedures.…”