Wehr attributed these 48-hour cycles to an "internal desynchronization" model wherein the temperature rhythm and many other circadian rhythms remained synchronized to the 24-hr environment, but the period of sleep-wake (and some associated rhythms) decelerated so much as to double cycle length and produce 48-hour rhythms.
The circadian clock provides the interface between an organism and its 24 hr geophysical environment.
Cells grew for 24 hr in this environment before being characterized via flow cytometry and diluted into the original environment they were in before the perturbation (hereafter, the 'recovery environment').
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Wildtype (WT) and homozygous mice from GluR1-S831A GluR1-S845A845A gene knockin lines (genetic background: C57BL6) [32], [32] were raised in a normal lighted environment (12 hr light/12 hr dark cycle) until postnatal age 21 days (P21).
Plates were incubated at 37 ± 1°C for 24 ± 1 hrs in gaseous environment of 5% ± 1% carbon dioxide.
The rats were housed 4 to a cage and maintained in a 12-hr light/dark cycle environment.
These doses would be equivalent to working 3 hr in the environments of boilermakers and welders with the ambient V and Zn concentrations of 8 and 10 μg/m, respectively (Marquart et al. 1989; Woodin et al. 2000), assuming ventilation of 10 L/min.
Male C57BL/6J mice were obtained from the Jackson Laboratory (Bar Harbor, ME), housed (4 per cage) in a controlled environment (12-hr light/12-hr dark daily cycle, 23 ± 1°C, 50 70% humidity) and fed ad libitium with standard chow (Harlan Tekladd, 2016) for 4 weeks prior to experimentation.
Mice were kept in a pathogen-free environment with 12 hr:12 hr light:dark cycle.
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