August 12, 2004 - A look inside a wristwatch reveals that timekeeping is a complex
affair, involving the coordination of mechanical parts providing the impulses and feedback
needed to achieve precisely recurring movement. Biological clocks are equally complex, regulated
by a network of genes and transcriptional factors that interact to stabilize the rhythms
of numerous physiological systems. Unlike the wristwatch, however, there is no visible readout
or display showing an individual's body time, a lack which has stood as one of the major
barriers to realizing the promise of chronotherapy, which seeks to deliver drug treatments
at optimal body times.
A new study by Hiroki R. Ueda (Laboratory for Systems
Biology, RIKEN Center for Developmental Biology;
Kobe, Japan) and colleagues has provided proof of principle that just such a display of individual body
time may one day be possible. The report, published in the August 3 issue of the Proceedings of the
National Academy of Sciences, describes the analysis of the expression of more than 100 time-indicating
genes in the mouse. The results of this genome-wide study enabled the authors to develop a "molecular
timetable" that provides an accurate representation of the animal's body time based on the sampling
of gene expression levels at a single point in time.
|Body time represented in a gene expression profile of time-indicating
genes. In the morning (center top) 'dawn-indicating genes' (green) are highly expressed, and 'dusk-indicating
genes' (pink) are expressed at low levels. In the evening (center bottom), their expression
levels are reversed.
Many genes exhibit variable expression cycles roughly over the course of a 24-hour day, a phenomenon
known as circadian rhythmicity. In their study, Ueda et al . first identified genes that are
expressed in high amplitude circadian patterns. Such genes demonstrate cyclical expression irrespective
of variations in exposure to light, maintaining their amplitude and periodicity even when the animals
are kept in constant darkness. The team identified 168 genes fulfilling these criteria in samples from
mouse liver, and calculated their peak expression times in terms of both external time and the animal's
subjective inner clock. They found that the genes could be organized into a kind of daily schedule by
their times of peak expression - some were expressed most highly at daybreak, some at dusk, some at
other time-points throughout the cycle of a single day.
By plotting the expression levels of the entire set of time-indicating genes against a 24-hour curve,
Ueda and colleagues found that it is possible to determine body time with a high degree of accuracy.
To test the robustness of this molecular time measurement system, they measured the expression levels
of the 168 genes at the lights-out phase in animals kept in conditions of alternating 12-hour periods
of light and dark. This phase is presumed to be a hotbed of circadian regulatory activity, as the mouse's
internal rhythm runs in cycles that are naturally shorter than 24 hours, and darkness serves as a stimulus
for resetting the circadian clock (such stimuli are referred to as Zeitgeber mechanisms, from
the German word meaning "time-giver"). Their analysis of expression levels at this critical Zeitgeber hour
12 produced estimated body times to an accuracy of about 1 hour in all animals studied, indicating that
the molecular timetable provided is strongly resistant to environmental noise (in this case, variation
in exposure to light).
They next tested the system in mice with a homozygous mutation in the gene Clock , whose loss
of function is known to perturb the natural circadian rhythm. They found rhythm disruptions in the expression
profiles of all of the Clock / Clock mice, suggesting that the molecular timetable
approach may also have applications in the diagnosis of circadian rhythm disorders.
This new approach to the horology of the body has been validated in mice of varying genetic
backgrounds and in principle can be applied to any organism exhibiting biological clock activity,
a range that spans the living world from bacteria to plants to humans. And indeed a test of the
method in wild type and Clock -mutant fruit flies gave similar results to those in the
The development by Ueda and colleagues of a universally-applicable, specific, sensitive and accurate
method capable of both detecting individual body time and of diagnosing circadian disorders
represents a major step toward the fulfillment of the longstanding dream of scientists and
physicians: to put a readable face on the body's clock. Its application in medical treatment may
one day allow doctors to tailor drug administration to a patient's body time, which promises to
optimize efficacy and reduce adverse effects.