While certain circadian hormonal changes are prominent, their predictable assessment requires a standardization of conditions of sampling. The 24-hour rhythm in circulating corticosterone of rodents, known since the 1950s, was studied as a presumed proxy for stress on 108 rats divided into 9 groups of 6 male and 9 groups of 6 female animals sampled every 4 hours for 24 hours. In a first stress study, the "no-rhythm" (zero-amplitude) assumption failed to be rejected at the 5% probability level in the two control groups and in 16 out of the 18 groups considered. A circadian rhythm could be detected with statistical significance, however, in three separate follow-up studies in the same laboratory, each on 168 rats kept on two antiphasic lighting regimens, with 4-hourly sampling for 7 or 14 days. In the first stress study, pooling of certain groups helped the detection and assessment of the circadian corticosterone rhythm. Without extrapolating to hormones other than corticosterone, which may shift more slowly or adjust differently and in response to different synchronizers, the three follow-up studies yielded uncertainty measures (95% confidence intervals) for the point estimate of its circadian period, of possible use in any future study as a reference standard. The happenstance of a magnetic disturbance at the start of two follow-up studies was associated with the detection of a circasemiseptan component, raising the question whether a geomagnetic disturbance could be considered as a "load". Far beyond the limitations of sample size, the methodological requirements for standardization in the experimental laboratory concerning designs of studies are considered in the context of models of depression. Lessons from nature's unforeseen geomagnetic contribution and from human studies are noted, all to support the advocacy, in the study of loads, of sampling schedules covering more than 24 hours.
Bibliographical noteFunding Information:
Supported by ETT 314/2006, Hungary; NIH GM-13981 (FH), and University of Minnesota Supercomput-ing Institute (GC, FH).
- Chronic mild stress
- Light shift