24-hr Clock Research Articles

Incremental Growth Lines in Mouse Molar Dentin Represent 8-hr Ultradian Rhythm.

Rhythmic incremental growth lines occur in dental hard tissues of vertebrates, and dentinogenesis in rodent incisors is suggested to be controlled by the 24-hr circadian clock. Rodent incisors continue to grow throughout the animal’s life; however, similar to human teeth, rodent molars stop growing after crown formation. This similarity suggests that the mouse molar is an excellent model to understand the molecular mechanisms underlying growth of human teeth. However, not much is known about the rhythmic dentinogenesis in mouse molars. Here, we investigated the incremental growth lines in mouse molar dentin using tetracycline as the growth marker. The incremental growth lines were observed to be generated at approximately 8-hr intervals in wild-type mice housed under 12:12 hr light-dark conditions. Moreover, the 8-hr rhythmic increments persisted in the wild-type and Bmal1−/− mice housed in constant darkness, where Bmal1−/− mice become behaviorally arrhythmic. These results revealed that the dentinogenesis in mouse molars underlie the ultradian rhythms with around 8-hr periodicity. Further, the circadian clock does not seem to be involved in this process, providing new insight into the mechanisms involved in the tooth growth.

A time to be born: Variation in the hour of birth in a rural population of Northern Argentina.

The present study aimed at investigating the timing of birth across the day in a rural population of indigenous and nonindigenous women in the province of Formosa, Argentina in order to explore the variation in patterns in a non-Western setting. This study utilized birth record data transcribed from delivery room records at a rural hospital in the province of Formosa, northern Argentina. The sample included data for Criollo, Wichí, and Toba/Qom women (n = 2421). Statistical analysis was conducted using directional statistics to identify a mean sample direction. Chi-square tests for homogeneity were also used to test for statistical significant differences between hours of the day. The mean sample direction was 81.04°, which equates to 5:24 AM when calculated as time on a 24-hr clock. Chi-squared analyses showed a statistically significant peak in births between 12:00 and 4:00 AM. Birth counts generally declined throughout the day until a statistically significant trough around 5:00 PM. This pattern may be associated with the circadian rhythms of hormone release, particularly melatonin, on a proximate level. At the ultimate level, giving birth in the early hours of the morning may have been selected to time births when the mother could benefit from the predator protection and support provided by her social group as well as increased mother-infant bonding from a more peaceful environment.

Developing a validated urinary diary: Phase 1

To develop and validate a urinary diary, using the psychometric validation protocol applied to all International Consultation on Incontinence Questionnaire (ICIQ) modules. Patient and clinician opinion was sought on diary content, format, and duration using interviews and questionnaires. Content deemed essential for inclusion were incorporated into four draft diary formats (Phase 1a). Four rounds of content validation using patient-completed diaries or patient interviews and one round of clinician opinion were undertaken, before a consensus was achieved (Phase 1b). Phase 1a: 27 patients were interviewed and 30 clinicians returned a questionnaire regarding diary content, format and duration. Content deemed essential for inclusion was identified. Patients reported no format preference but preferred a diary of ≤7 days. Clinicians preferred a 3-day diary in single sheet format. Phase 1b: 75 of 200 (37.5%) patients returned completed draft diaries. Equal preference for the two portrait format diaries was expressed. Twenty-six clinicians returned a questionnaire regarding their preference for either of these two options, with 69% (18/26) selecting the one with a printed 24-hr clock. Optimal diary duration was 4 days. Frequency, voided volume, fluid intake (amount, time and type), time of incontinence episode, and bladder sensation were supported for inclusion in the final diary. Diary content, format, and duration deemed essential by both patients and clinicians have been identified in this preliminary phase, allowing the formation of a draft urinary diary. Further tests of diary validity, reliability and responsiveness will be evaluated in Phase 2 of the study.

Assessment of Circadian and Light-Entrainable Parameters in Mice Using Wheel-Running Activity.

In most organisms, physiological variables are regulated by an internal clock. This endogenous circadian (∼24-hr) clock enables organisms to anticipate daily environmental changes and modify behavioral and physiological functions appropriately. Processes regulated by the circadian clock include sleep-wake and locomotor activity, core body temperature, metabolism, water/food intake, and available hormone levels. At the core of the mammalian circadian system are molecular oscillations within the hypothalamic suprachiasmatic nucleus. These oscillations are modifiable by signals from the environment (so called zeitgebers or time-givers) and, once integrated within the suprachiasmatic nucleus, are conveyed to remote neural circuits where output rhythms are regulated. Disrupting any of a number of neural processes can affect how rhythms are generated and relayed to the periphery and disturbances in circadian/entrainment parameters are associated with numerous human conditions. These non-invasive protocols can be used to determine whether circadian/entrainment parameters are affected in mouse mutants or treatment groups. Curr. Protoc. Mouse Biol. 1:369-381 © 2011 by John Wiley & Sons, Inc.

The organization of the suprachiasmatic circadian pacemaker of the rat and its regulation by neurotransmitters and modulators.

The long-term goal of our research is to understand how cells of the suprachiasmatic nucleus (SCN) are organized to form a 24-hr biological clock, and what roles specific neurotransmitters and modulators play in timekeeping and resetting processes. We have been addressing these questions by assessing the pattern of spontaneous neuronal activity, using extracellular and whole-cell patch recording techniques in long-lived SCN brain slices from rats. We have observed that a robust pacemaker persists in the ventrolateral region of microdissected SCN, and have begun to define the electrophysiological properties of neurons in this region. Furthermore, we are investigating changing sensitivities of the SCN to resetting by exogenous neurotransmitters, such as glutamate, serotonin, and neuropeptide Y, across the circadian cycle. Our findings emphasize the complexity of organization and control of mammalian circadian timing.

Minute-by-minute analysis of REM sleep timing in major depression

Sleep changes described in depressed patients may represent alterations in the timing of rapid-eye-movement (REM) sleep or sleep onset. We examined these variables in groups of healthy control subjects ( n = 47), depressed outpatients ( n = 98), and depressed inpatients ( n = 41). Outpatients depressives had greater severity of clinical symptoms than inpatients using the Hamilton Rating Scale for Depression. The depressed inpatient group had a later mean sleep onset time than the other groups, and the depressed outpatient group had a wider range of good night times than control subjects. REM timing in each group was examined as a relative frequency distribution of REM sleep (FDRS) for each minute across the night. The FDRSs for the three groups were statistically compared using the parameters from a two-component model, which includes a deterministic sinusoidal function and a time series process for errors. The slope of the linear trend in the FDRS rhythm was smaller (less positive) for both depressed groups than for controls. The ultradian FDRS rhythm occurred at an earlier phase, relative to sleep onset, in the inpatient depressed group compared to the control group. The ultradian FDRS rhythm had a longer period in the outpatient group compared to the control and inpatient groups. When referenced to 24-hr clock time in an exploratory analysis, the depressed groups appeared to have less robust FDRS ultradian rhythms than controls, but they did not appear to have a systematic phase alteration compared to controls. Abnormalities of REM sleep timing in groups of depressed patients may reflect a disturbance of sleep initiation and generation, or difficulty in entrainment of REM, rather than a systematic phase alteration in REM sleep propensity.

"Dark-active" rat transformed into "light-active" rat by destruction of 24-hr clock: function of 24-hr clock and synchronizers.

In alternating 12-hr periods of light and dark the rat is active mainly in the dark. Its activity in the dark (beginning at 1800) depends exclusively on release of activity by the 24-hr clock. In the light (beginning at 0600) the 24-hr clock inhibits activity; the normal rat becomes totally inactive in the light except for activity resulting from external stimulation. After section of the connections between the optic chiasma and the hypothalamus, some rats become totally and permanently inactive in the dark. This sectioning destroys the 24-hr clock. After destruction of the clock removes inhibition of activity in the light period, the rat becomes active promptly at start of the light period--i.e., becomes a "light-active" animal. In the normal rat, activity becomes synchronized to start of the dark (by the electric clock at 1800), regardless of the amounts of activity. Destruction of the 24-hr clock eliminates the synchronizer at 1800. However, almost at once, activity, eating, and drinking are kept together by a second synchronizer, start of the light (by the electric clock at 0600). This may explain the ability of the rat to survive after destruction of the 24-hr clock.

Evidence for existence of a yearly clock in surgically and self-blinded chipmunks.

By use of simplified technique and constant environmental conditions, I have demonstrated the existence in the chipmunk of a yearly clock. In a blinded chipmunk the clock manifested itself by remarkably consistent changes in running activity, food and water intake, and body weight over 6 1/2 yr. Studies on freshly trapped chipmunks kept in the same laboratory environment but with alternating light and darkness (12 hr each), showed that, when their eyes were covered for much of the light period, they reduced their exposure to light to preserve the activity of the yearly clock. Laboratory-adapted chipmunks that do not shield their eyes from light do not show the clock. The yearly clock has all the characteristics of the 24-hr clock, including sharply defined active and inactive phases, and must likewise play an important part in the animal's survival. Light would appear to be the chief or only cue for the clock. The period lengths did not change with age during the 6 1/2 yr.

Heavy water as a tool for study of the forces that control length of period of the 24-hour clock of the hamster.

In alternating 12-hr periods of light and darkness, start of the dark period entrains the hamster's 24-hr clock. Blinding or constant darkness frees the clock of entrainment by allowing it to run faster or slower than 24 hr. Constant light frees the clock from entrainment and permits it to run slower than 24 hr--that is, lengthening its period. Heavy water given in drinking water linearly lengthens the period of the 24-hr clock of blinded hamsters or of hamsters kept in constant darkness in direct proportion to concentration of heavy water (1--50%). Heavy water (1-35%) has very different effects on length of the periods of the 24-hr clock when given under conditions of alternating 12-hr periods of light and darkness. Under these conditions, length of the period is controlled by three factors: (i) heavy water which slows the 24-hr clock; (ii) constant light which also slows the 24-hr clock; (iii) the counteracting effects of entrainment. It is thus possible to observe the effects of all three forces simultaneously in the same animal. The clock slowed by heavy water (1--20%) showed a strong tendency to return to a 24-hr entrainment whenever possible. On a 50% concentration of heavy water, the length of period of the clock became markedly lengthened but very constant and apparently independent of all external and internal disturbances.

Continuous light or darkness and circadian periodic mitosis and metabolism in C and D8 mice.

In immature C mice exposed first to alternating 12 hr of light and 12 hr of darkness (LD), and maintained thereafter in constant darkness for several days, the circadian rhythms in hepatic and pinnal mitosis are demonstrable by spot checks made at the approximate times of LD-synchronized peak and trough. Spot checks at same times in mice of same stock and age, kept for several days in constant light, reveal the cell division rhythm of liver parenchyma, but not that of pinnal epidermis. In immature D8 mice kept for several days in constant darkness, rhythms in hepatic mitosis, phospholipid, ribonucleic and deoxyribonucleic acid metabolism persist, while cell division rhythm in ear pinna of same animals is not detectable with the particular spot check used. In mice of same stock and age, on the 4th day in constant light, a significant rhythm persists in the relative specific activity of the hepatic phospholipid; timing of this metabolic cellular rhythm is drastically desynchronized from the reference standard of a 24-hr clock. Data reveal persistence of some mitotic and metabolic circadian rhythms under conditions studied, with phase drifts or phase shifts of these rhythms occurring both in relation to the 24-hr clock and among the rhythms themselves. These changes in external and internal timing of a circadian system are more extensive in constant light than in constant darkness.