Cortisol-mediated synchrinization of circadian rhythm in urinary potassium excretion.

Conscious chair-acclimatized squirrel monkeys (Saimiri sciureus) studied with lights on (600 lx) from 0800 to 2000 h daily (LD 12:12) display a prominent circadian rhythm in renal potassium excretion. The characteristics of this rhythm were reproduced in adrenalectomized monkeys by infusing 5 mg cortisol and 0.001 mg aldosterone, or 5 mg cortisol alone, between 0800 and 0900 h daily. When the timing of cortisol adminisration (with or without aldosterone) was phase-delayed by 8 h, the urinary potassium rhythm resynchronized by 80% of the cortisol phase shift, but only after a transient response lasting 3-4 days. With the same daily dose of adrenal steroids given as a continuous infusion throughout each 24 h, urinary potassium excretion showed free-running oscillations no longer synchronized to the light-dark cycle. These results indicate that the cirdacian rhythm of plasma cortisol concentration acts as an internal mediator in the circadian timing system, synchronizing a potentially autonomous oscillation in renal potassium excretion to environmental time cues and to other circadian rhythms within the animal.     

Circadian rhythms in the flow rate and composition of unstimulated and stimulated human submandibular saliva.

1. Nine subjects recorded oral temperature and collected unstimulated submandibular saliva and submandibular saliva stimulated by sour lemon drops at a constant flow rate of 1 ml./min, at about 07.00, 11.00, 14.00, 17.00 and 22.00 h daily for time spans of about 11 days. 2. Least-squares cosine waves were fitted to the data to test for the presence and characteristics of circadian rhythms and the results were subjected to cosinor analysis and the Rayleigh test to assess the statistical significance of any rhythms for the group of subjects as a whole (i.e. whether or not the computed acrophases were randomly distributed throughout the 24 h of the day). 3. Unstimulated submandibular saliva showed circadian rhythms, significant for the group as a whole, in flow rate and in the concentrations of sodium, potassium, magnesium, chloride and inorganic phosphate but not in protein or calcium. 4. Stimulated submandibular saliva showed circadian rhythms, significant for the group as a whole, in sodium, potassium, calcium, magnesium, chloride and inorganic phosphate but not in protein. Oral temperature also showed a significant circadian rhythm. 5. Because of the high amplitude of many of these rhythms, they must be taken into account when establishing the normal range of salivary values, or when salivary composition is to be used as an indicator of systemic disease or to be implicated in the aetiology of oral disease.     

Suprachiasmatic nuclei lesions do not eliminate the circadian rhythms of electrolyte excretion in the rat

The 24-hour variations in 2-hour diuretic and saluretic action of furosemide (4 mg/kg) given at 8, 12, 16, 20, 24 or 4 hr and the circadian rhythms of food and water intake, urine and electrolyte excretion followed for two consecutive days at 4-hour intervals were investigated in suprachiasmatic nuclei (SCN)-lesioned or sham-operated male Wistar rats. The results showed that: a distinct 24-hour rhythm in furosemide-induced urine, sodium and chloride excretion persisted after SCN lesions, and the lesions abolished the circadian rhythms of food and water intake but only desynchronized the individual 24-hour variations in electrolyte excretion. We concluded that SCN play a role as a central synchronizer but not as a major oscillator of the circadian rhythms of electrolyte excretion in the rat.     

Circadian rhythms in human salivary flow rate and composition

1. Unstimulated whole saliva and parotid saliva stimulated at a constant flow rate of 1.0 ml./min were collected from eight subjects at about 07.00, 11.00, 14.00, 17.00 and 22.00 hr and oral temperature was recorded several times daily for time spans of between 4 and 26 days. A least-squares cosine wave was fitted to the data to test for the presence and characteristics of circadian rhythms.2. Estimates of mean level, amplitude, acrophase and period were obtained for different components and the results were subjected to cosinor analysis.3. Unstimulated whole saliva showed significant circadian rhythms in flow rate and in the concentrations of sodium and chloride but not in protein, potassium, calcium, phosphate or urea.4. Stimulated parotid saliva showed significant circadian rhythms in the concentrations of protein, sodium, potassium, calcium and chloride but not in phosphate or urea5. Oral temperature showed a circadian rhythm which, like the salivary rhythms, was of a 24.0 hr periodicity.     

Transient circadian internal desynchronization after light-dark phase shift in monkeys.

In four conscious chair-acclimatized squirrel monkeys (Saimiri sciureus) studied with lights on (600 lx) from 0800 to 2000 h daily (LD 12:12), prominent 24-h rhythms in feeding, drinking, activity, body temperature, and urinary potassium, sodium, and water excretion were seen. When the monkeys were subjected to 36 h of darkness followed by 36 h of light each variable demonstrated a circadian rhythm which was not passively dependent on the light-dark cycle. After the 24-h light-dark cycle was abruptly phase-delayed by 8 h, all the rhythms resynchronized with the new light-dark cycle phase, demonstrating that light-dark cycles are an effective zeitgeber. However, the resynchronization of the rhythms of feeding, drinking, activity, and body temperature was 90% complete within approximately 2 days while the 90% resynchronization of the urinary rhythms took approximately 5 days. These results suggest that the circadian timing system in S. sciureus may consist of several spontaneously oscillating units which can become transiently uncoupled during pertubations of environmental time cues.     

A mammalian ortholog of Drosophila timeless, highly expressed in SCN and retina, forms a complex with mPER1

BACKGROUND: It is now becoming clear that the circadian rhythm of behaviours and hormones arises from a rhythm at the level of gene expression, and that mammals and Drosophila essentially use homologous genes as molecular gears in the control of circadian oscillation. In Drosophila, the period and timeless genes form a functional unit of the clock and its autoregulatory feedback loop for circadian rhythm. However, in mammals, the counterpart of timeless has not been found. RESULTS: We have isolated a mammalian homologue of timeless, mTim, from the mouse brain. mTim is highly expressed, with a weak or absent rhythm in the suprachiasmatic nucleus, the mammalian circadian centre. In the retina, mTim mRNA was found to be expressed with a circadian rhythm, and a particularly robust cycle was observed in the presence of light/dark cycles. We demonstrated that mTIM physically associates with mPER1 in vitro and in the nuclei of cultured COS7 cells. CONCLUSIONS: We have reported the isolation of the mouse timeless cDNA, the expression of the mTim mRNA and an interaction of mTIM with mPER1. These results indicate that the autoregulatory feedback mechanism of circadian oscillation of the period gene may also be conserved in mammals.     

Effects of mineralocorticoids on Na+ and K+ excretion in the adrenalectomized rat

The short-term effects (within 4 h) of low doses of intravenous aldosterone or deoxycorticosterone on potassium and sodium urinary excretion were studied by clearance techniques in 24-h adrenalectomized, anesthetized male rats. All animals were substituted with a glucocorticoid (dexamethasone; plasma concentration approximately 6 nM) to maintain normal glomerular filtration rate. The mineralocorticoid effects were studied under various conditions of sodium and potassium load. Mineralocorticoid administration uniformly resulted in antinatriuresis, starting within 30-60 min and, at the peak effect, amounting to 1-2% sodium fractional excretion. The level of antinatriuresis was directly related to the control sodium excretion before aldosterone administration. Mineralocorticoids induced a significant kaliuresis in all groups except one, the one receiving the lowest sodium load. The aldosterone-induced kaliuresis was also related to the sodium load and the control fractional sodium excretion level and was simultaneous with the beginning of the reduced sodium excretion. In control, mineralocorticoid-deprived rats, kaliuresis was not enhanced by increasing the sodium load. Control as well as mineralocorticoid-treated rats responded by an increased kaliuresis following an acute potassium load and by a decreased kaliuresis after 3 days of low potassium diet.     

The effect of real and simulated time-zone shifts upon the circadian rhythms of body temperature, plasma 11-hydroxycorticosteroids, and renal excretion in human subjects

1. Observations were made upon five subjects who flew through 4(1/2)-6 time zones, four of them returning later to their starting point, and upon twenty-three subjects experiencing simulated 6 or 8 hr time zones shifts in either direction in an isolation unit.2. Measurements were made of plasma concentration of 11-hydroxycorticosteroids, of body temperature, and of urinary excretion of sodium, potassium and chloride. Their rhythm was defined, where possible, by fitting a sine curve of period 24 hr to each separate 24-hr stretch of data and computing the acrophase, or maximum predicted by the sine curve.3. The adaptation of the plasma steroid rhythm was assessed by the presence of a sharp fall in concentration after the sample collected around 08.00 hr. The time course of adaptation varied widely between individuals; it was usually largely complete by the fourth day after westward, and rather later after eastward, flights. After time shift the pattern often corresponded neither to an adapted nor to an unadapted one, and in a subject followed for many months after a real flight a normal amplitude only appeared 2-3 months after flight.4. Temperature rhythm adapted by a movement of the acrophase, without change in amplitude, although on some days no rhythm could be observed. This movement was always substantial even on the first day, and was usually nearly complete by the fifth.5. High nocturnal excretion of electrolyte was often seen in the early days after time shift, more notably after simulated westward flights. Adaptation of urinary electrolyte rhythms usually proceeded as with temperature, but the movement of the acrophase was slower, more variable between individuals, more erratic, and sometimes reversed after partial adaptation. On a few days there were two maxima corresponding to those expected on real and on experimental time.6. Sodium excretion was much less regular than that of potassium, but adapted more rapidly to time shift, so that the two often became completely dissociated. Chloride behaved much as sodium.7. The time course of adaptation of the plasma steroid and urinary potassium rhythms were sufficiently similar to suggest a causal connexion. The time course of adaptation of the temperature rhythm did not coincide with that of any other component considered here.     

Circadian periodicity of brain activity and urinary excretion in the epileptic baboon

A 24-H rhythm in the proclivity of the baboon Papio papio to exhibit a seizure response to intermittent light stimulation (ILS) has been uncovered. Seizure response to ILS was found to be greatest in the morning at about 0900 h, 2 h after lights on, and least severe in the evening at about 2000 h 1 h after lights off. Urine cortisol and potassium excretion rhythms followed the same pattern as seizure susceptibility and sodium rhythms were 3 or 4 delayed. Mean values for urine cortisol per 4 h ranged from 25 microgram/4 h at the maxima (0400 to 0800 h) to 2 microgram/4 h at the minima (2000 to 2400 h). Electrolyte rhythms ranged from 13 to 3 meq/4 h for potassium and 1 to 5 meq/4 h for sodium. Changes in electroencephalographic (EEG) activity also accompanied the change in urine cortisol and potassium, and seizure sensitivity, Changes in the EEG at the diurnal minima (2000 h) consisted of a decrease in slow waves concomitant with an increase in fast activity in the 18-25 Hz range,     

Sleep and stressor exposure in relation to circadian rhythms in catecholamine excretion.

12 healthy male volunteers spent 64 h of continuous waking under strictly controlled environmental conditions (light, food, drink, activity) in isolation from the external world. Before and after the vigil the subjects slept in the laboratory. An additional group of five participated only during day time and spent the intervening night period asleep at home. Measurements were carried out in 3 h intervals except for sleep periods. Shortly before the circadian trough and peak of adrenaline excretion respectively, the subjects were exposed to a performance stressor. Results from the vigil showed a very pronounced circadian rhythm for adrenaline excretion but none for noradrenaline excretion. For adrenaline, night-time sleep reduced excretion levels, causing an even more pronounced circadian pattern. For noradrenaline, night-time sleep caused a drop in excretion giving the impression of a circadian rhythm. These and previous results led to the conclusion that the excretion of adrenaline exhibits a self-sustained rhythm while a rhythm in noradrenaline excretion is found only when caused by external synchronizers such as sleep-wake alternation. No difference in magnitude of stress response between peak and trough was observed for any of the catecholamines. Night-time (trough) exposure completely obliterated the circadian rhythm of adrenaline excretion for the duration of the exposure. It was concluded that the normal pronounced night-time trough of adrenaline cannot be due to unavailability of adrenaline in the medulla. With respect to sleep deprivation, no effect was found on excretion levels during waking, during sleep, or in response to the stressor as deprivation progressed. Finally, there was also found to be a close temporal covariation between adrenaline excretion and both rectal temperature and self-rated fatigue (neg.).     

Daytime Exposure to Blue Light Alters Cardiovascular Circadian Rhythms,Electrolyte Excretion and Melatonin Production

Artificial light is characterized by certain features of its impact on the body in terms of its spectral distribution of power, duration of exposure and intensity. Short waves, perceived as blue light, are the strongest synchronizing agent for the circadian system. In the present work, we investigated the features of the circadian rhythms of blood pressure (BP), heart rate (HR), the excretion of electrolytes and the secretion of melatonin in normotensive (Wistar–Kyoto) and hypertensive (SHR) rats under the action of monochromatic blue light in the daytime period. It was found that the exposure of Wistar–Kyoto rats to monochromatic blue light was accompanied by a significant decrease in nighttime and 24 h systolic BP. The most remarkable changes are characteristic of the HR in SHR rats under monochromatic light. A significant decrease in HR in each time period was found, but the predominance of nighttime over daytime values remained in SHR animals. There was also a significant increase in the mesor of the HR in SHR rats. Additionally, the amplitude of diastolic BP and HR, as well as the range of oscillations in HR, were significantly increased compared with the standard light pattern. In contrast to SHR rats, the regulation of the circadian rhythms in Wistar–Kyoto rats was more flexible and presented more changes, which may be aimed at the adaptation of the body to environmental conditions. For Wistar–Kyoto rats, an increase in the level of excreted electrolytes was observed under the action of monochromatic light, but no similar changes were found in SHR rats. For Wistar–Kyoto rats, a significant decrease in the urine concentration of aMT6s in the daytime and nighttime periods is characteristic, which results in the loss of the circadian rhythm. In SHR rats, there was a significant decrease in the nighttime content of aMT6s in the urine, while the daytime concentration, on the contrary, increased. The obtained data demonstrate that prolonged exposure to monochromatic blue light in the daytime period affects the circadian structure of the rhythms of the cardiovascular system, the rhythm of electrolyte excretion and the production of epiphyseal melatonin in wild-type and hypertensive animals. In SHR rats, the rhythms of BP and HR exhibit a more rigid pattern.     

Effects of water diuresis and osmotic (mannitol) diuresis on urinary solute excretion by the conscious rat

1. The time course and extent of changes in urinary flow and in the outputs of urea, Na(+), K(+), and NH(4) (+) over a period of 7(1/2) hr in conscious rats during water and osmotic (mannitol) diuresis were determined, and compared with spontaneous changes in non-diuretic animals.2. In non-diuretic rats, a morning rise and subsequent decline in urinary osmolal, sodium, potassium and ammonium outputs occurred, possibly attributable to circadian rhythms.3. Water diuresis was accompanied by (i) a rapid increase in urea excretion during the phase of increasing urine flow, followed by a fall in later periods to values similar to those in non-diuresis, (ii) a slower increase in sodium output, continuing after the establishment of the constant water load, (iii) unchanged potassium excretion, but slightly increased ammonium outputs.4. Mannitol diuresis was accompanied by (i) a rapid increase in urea outputs which subsequently fell but remained significantly higher, (ii) a steep rise in sodium and potassium outputs to values which remained far higher than those in non-diuretic and water diuretic animals.5. The changes in mannitol diuresis are considered to result mainly from decreased tubular reabsorption, due to the lowered intraluminal sodium, potassium and urea concentrations and increased intratubular fluid flow. Some of the acute increase in urea excretion may be due to washout of medullary urea into the tubular fluid.6. In water diuresis, some of the changes in solute excretion may similarly result from altered tubular reabsorption, perhaps influenced by suppression of anti-diuretic hormone (A.D.H.). In addition, the slower changes in sodium output may be related to several consequences of change in body fluid volume.     

Circadian rhythm of core body temperature in subjects with chronic fatigue syndrome.

The pathophysiological basis for chronic fatigue syndrome (CFS) remains poorly understood. Certain symptoms of CFS, namely fatigue, neurocognitive symptoms and sleep disturbance, are similar to those of acute jet lag and shift work syndromes thus raising the possibility that CFS might be a condition associated with disturbances in endogenous circadian rhythms. In this study, we tested this hypothesis by examining the circadian rhythm of core body temperature (CBT) in CFS and control subjects. Continuous recordings of CBT were obtained every 5 min over 48 h in a group of 10 subjects who met the Center for Disease Control (CDC) definition of CFS and 10 normal control subjects. Subjects in the two groups were age, sex and weight-matched and were known to have normal basal metabolic rates and thyroid function. CBT recordings were performed under ambulatory conditions in a clinical research centre with the use of an ingestible radio frequency transmitter pill and a belt-worn receiver-logger. CBT time series were analysed by a cosinor analysis and by a harmonic-regression-plus-correlated-noise model to estimate the mean, amplitude and phase angle of the rhythm. The goodness of fit of each model was also compared using the Akaike Information Criterion (AIC) and sigma2. Average parameters for each group were compared by Student's t-test. By cosinor analysis, the only significant difference between CFS and control groups was in the phase angle of the third harmonic (P=0.02). The optimal harmonic-regression-plus-correlated-noise models selected were ARMA(1,1): control 7, CFS 6; ARMA(2,0): control 1, CFS 4; and ARMA(2,1): control 2 subjects. The optimal fit ARMA model contained two harmonics in eight of 10 control subjects but was more variable in the CFS subjects (1 harmonic: 5 subjects; 2 harmonics: 1 subject; 3 harmonics: 4 subjects). The goodness of fit measures for the optimal ARMA model were also better in the control than the CFS group, but the differences were not statistically significant. We conclude that, measured under ambulatory conditions, the circadian rhythm of CBT in CFS is nearly indistinguishable from that of normal control subjects although there was a tendency for greater variability in the rhythm. Hence, it is unlikely that the symptoms of CFS are because of disturbance in the circadian rhythm of CBT.     

Metabolic heat production,heat loss and the circadian rhythm of body temperature in the rat

Metabolic heat production (calculated from oxygen consumption), dry heat loss (measured in a calorimeter) and body temperature (measured by telemetry) were recorded simultaneously at 6 min intervals over five consecutive days in rats maintained in constant darkness. Robust circadian rhythmicity (confirmed by chi square periodogram analysis) was observed in all three variables. The rhythm of heat production was phase-advanced by about half an hour in relation to the body temperature rhythm, whereas the rhythm of heat loss was phase-delayed by about half an hour. The balance of heat production and heat loss exhibited a daily oscillation 180 deg out of phase with the oscillation in body temperature. Computations indicated that the amount of heat associated with the generation of the body temperature rhythm (1.6 kJ) corresponds to less than 1 % of the total daily energy budget (172 kJ) in this species. Because of the small magnitude of the fraction of heat balance associated with the body temperature rhythm, it is likely that the daily oscillation in heat balance has a very slow effect on body temperature, thus accounting for the 180 deg phase difference between the rhythms of heat balance and body temperature.     

Neural regulation of the hepatic circadian rhythm

A microarray analysis experiment has revealed that there are many genes, including so-called clock genes, expressing a circadian rhythm in the liver. The clock genes mentioned above are expressed not only in the suprachiasmatic nucleus (SCN) of the hypothalamus, where the master clock exists, but also in other brain regions and various peripheral tissues. In the liver, clock genes are abundantly expressed and show a clear circadian rhythm. Thus, clock genes seem to play a critical role in the molecular clockworks of both the SCN and the liver. Although oscillation of clock genes in the liver is controlled under the circadian clock mechanism in the SCN, we do not know the resetting signals on liver clock function. Over the past few years, use of the pseudorabies virus, a transsynaptic tract tracer, has allowed us to map neural connections between the SCN and peripheral tissues in several physiological systems. Communication between the SCN and peripheral tissues occurs through autonomic nervous systems involving the sympathetic and parasympathetic neurons. This review mainly describes both anatomical and physiological experiments to reveal the sympathetic control over liver clock function. Although further study is necessary to produce the precise mechanism underlying neural control of liver clock systems, evolution of this mechanism will help our understanding of liver clock functions such as drug metabolism and energy metabolism.     

Role of the renin-angiotensin system in potassium control

To determine the importance of the renin-angiotensin system in control of plasma potassium concentration and excretion, potassium control was studied in two groups of dogs in response to a 20-fold increase in sodium intake (from 10 to 200 meq/day). Group I was intact whereas group II lacked feedback control of the renin-angiotensin system, which was eliminated by continuous infusion of 10 ng . kg-1 . min-1 angiotensin II. This rate of infusion reduced endogenous plasma renin activity (PRA) to undetectable levels throughout the study. The sodium forcing did not result in measurable changes in plasma potassium concentration or excretion in group I, in which PRA fell to 40% and plasma aldosterone concentration (PAC) to 60% of the low sodium levels. In group II the same sodium forcing produced a 12% decrease in plasma potassium concentration and a 79% increase in urinary potassium excretion. PAC also fell to 60% of the low sodium level in group II. The results demonstrate the importance of the renin-angiotensin system as a link between the nephron and the zona glomerulosa that is essential in controlling plasma potassium concentration and excretion during changes in sodium balance.     

Dependency of renal potassium excretion on Na,K-ATPase transport rate

Potassium secretion may depend on the transport rate of Na, K-ATPase in basolateral cell membranes of distal tubular cells. To examine this hypothesis experiments were performed in anaesthetized dogs during inhibition of proximal potassium reabsorption by acetazolamide or mannitol (fractional potassium excretion 1.2-1.4) or additional stimulation of potassium secretion by ethacrynic acid (fractional potassium excretion 2.1). Ouabain in a dose which inhibits 70–80% of the Na, K-ATPase activity reduced fractional potassium excretion to 0.8-0.9 by an effect on distal tubular secretion since potassium transport in the proximal tubules was not affected. Ouabain-sensitive potassium excretion varied in proportion to ouabain-sensitive sodium reabsorption during variation in glomerular nitration rate, even at urinary sodium concentrations exceeding 80 mmol 1^-1. In experiments without ouabain, saline infusion raised potassium excretion and sodium reabsorption until maximal Na, K-ATPase transport rate was reached, as judged from heat production measurements, but not during further increments in urine flow. After inhibition of Na, K-ATPase activity by hypokalaemia, potassium excretion and cortical heat production remained constant over a wide range of urine flow and sodium excretion. We conclude that potassium secretion is dependent on intact Na, K-ATPase activity and is stimulated by sodium delivery to the distal nephron until maximal transport rate of the enzyme is reached.     

Dependency of renal potassium excretion on Na,K-ATPase transport rate

Potassium secretion may depend on the transport rate of Na, K-ATPase in basolateral cell membranes of distal tubular cells. To examine this hypothesis experiments were performed in anaesthetized dogs during inhibition of proximal potassium reabsorption by acetazolamide or mannitol (fractional potassium excretion 1.2 - 1.4) or additional stimulation of potassium secretion by ethacrynic acid (fractional potassium excretion 2.1). Ouabain in a dose which inhibits 70-80% of the Na, K-ATPase activity reduced fractional potassium excretion to 0.8 - 0.9 by an effect on distal tubular secretion since potassium transport in the proximal tubules was not affected. Ouabain-sensitive potassium excretion varied in proportion to ouabain-sensitive sodium reabsorption during variation in glomerular filtration rate, even at urinary sodium concentrations exceeding 80 mmol X 1(-1). In experiments without ouabain, saline infusion raised potassium excretion and sodium reabsorption until maximal Na,K-ATPase transport rate was reached, as judged from heat production measurements, but not during further increments in urine flow. After inhibition of Na,K-ATPase activity by hypokalaemia, potassium excretion and cortical heat production remained constant over a wide range of urine flow and sodium excretion. We conclude that potassium secretion is dependent on intact Na,K-ATPase activity and is stimulated by sodium delivery to the distal nephron until maximal transport rate of the enzyme is reached.