Rhythm Of Plasma Corticosterone Research Articles

Misaligned feeding schedule elicits divergent circadian reorganizations in endo- and exocrine pancreas clocks.

Misaligned feeding may lead to pancreatic insufficiency, however, whether and how it affects circadian clock in the exocrine pancreas is not known. We exposed rats to a reversed restricted feeding regimen (rRF) for 10 or 20days and analyzed locomotor activity, daily profiles of hormone levels (insulin, glucagon, and corticosterone) in plasma, and clock gene expression in the liver and endocrine and exocrine pancreas. In addition, we monitored responses of the exocrine pancreatic clock in organotypic explants of mPer2Luc mice in real time to acetylcholine, insulin, and glucocorticoids. rRF phase-reversed the clock in the endocrine pancreas, similar to the clock in the liver, but completely abolished clock gene rhythmicity and significantly downregulated the expression of Cpb1 and Cel in the exocrine pancreas. rRF desynchronized the rhythms of plasma insulin and corticosterone. Daily profiles of their receptor expression differed in the two parts of the pancreas and responded differently to rRF. Additionally, the pancreatic exocrine clock responded differently to treatments with insulin and the glucocorticoid analog dexamethasone in vitro. Mathematical simulation confirmed that the long-term misalignment between these two hormonal signals, as occurred under rRF, may lead to dampening of the exocrine pancreatic clock. In summary, our data suggest that misaligned meals impair the clock in the exocrine part of the pancreas by uncoupling insulin and corticosterone rhythms. These findings suggest a new mechanism by which adverse dietary habits, often associated with shift work in humans, may impair the clock in the exocrine pancreas and potentially contribute to exocrine pancreatic insufficiency.

Effect of cold and warm white light on selected endocrine and immune parameters of broiler embryos and hatchlings

Lighting conditions during incubation can influence embryonic development, post-hatching ontogeny and production efficiency. Previous studies revealed that different light colours differently affect pineal melatonin biosynthesis in embryos and postembryonic development of broiler chickens, but physiological mechanisms mediating these effects are not known. Cold and warm white light consists of different wavelengths and therefore the aim of the present study was to explore if these two lights can differently influence the development of circadian melatonin biosynthesis, production of thyroid hormones and corticosterone, concentration of metabolites (glucose, cholesterol and triacylglycerols) as well as expression of two important immune genes, presenilin 1 and avian beta-defensin 1 (AvBD-1). We evaluated these traits in embryos before hatching (456, 460, 465, 468 and 472 h of incubation) and in hatchlings. The rhythmic profile of pineal melatonin with higher concentrations during the dark time was determined in both treatment groups. Melatonin levels increased considerably in hatchlings in comparison with embryos, but we found no difference in rhythm characteristics between groups. We did not identify any daily rhythms in plasma corticosterone and thyroid hormone levels in either studied age and no differences were found between light treatments in concentrations of thyroid hormones, corticosterone, metabolites and expression of presenilin and AvBD-1. The expected developmental increase of thyroid hormones was proved. Gene expression of presenilin increased in the duodenum of hatchlings in comparison with embryos, but the expression did not change in the bursa of Fabricius. On the other hand, expression of AvBD-1 decreased in hatchlings compared to embryos in both tissues. Based on these results, we can conclude that the colour temperature of white light did not influence endocrine and immune parameters determined in this study and probably monochromatic rather than polychromatic light should be used to influence embryonic development and postembryonic ontogeny of broiler chickens.

Abstract # 3188 Spinal cord injury perturbs circadian rhythms

Spinal cord injury (SCI) can disrupt many physiological functions. The circadian system helps maintain homeostasis throughout the body by regulating daily rhythms in physiology and behavior. Circadian rhythms likely exist in all cell throughout the body, yet whether SCI alters daily rhythms remains under-studied. Here, we hypothesized that SCI in rats would disrupt several prominent circadian outputs including glucocorticoids, core temperature, activity, and circadian/inflammatory gene expression. Female and male rats received clinically relevant thoracic (T-) 8 moderate-to-severe contusion SCI (or laminectomy sham surgery). Circadian measures – including rhythms of plasma corticosterone, body temperature and activity (using small implanted transmitters), plasma glucose, and gene expression – were studied before and after surgery. First, we found that SCI disrupted intraspinal and peripheral rhythms of clock and inflammatory gene expression. Circadian rhythms in peripheral cells are entrained by “oscillatory factors”, including glucocorticoids, body temperature, and activity. SCI increased and disrupted rhythms of the major rodent glucocorticoid, corticosterone. Pre-surgery and sham rats displayed expected rhythms in activity and body temperature, whereas SCI rats had blunted daily rhythms in activity and body temperature. In parallel, SCI increased plasma glucose and liver expression of glucose metabolism genes. Our data show that moderate SCI in rats causes wide-ranging circadian dysfunction that is severe at acute time points and gradually recovers. Normalizing post-SCI circadian rhythms could enhance recovery of homeostasis and quality-of-life.

Spinal Cord Injury in Rats Disrupts the Circadian System.

Spinal cord injury (SCI) perturbs many physiological systems. The circadian system helps maintain homeostasis throughout the body by synchronizing physiological and behavioral functions to predictable daily events. Whether disruption of these coordinated daily rhythms contributes to SCI-associated pathology remains understudied. Here, we hypothesized that SCI in rats would dysregulate several prominent circadian outputs including glucocorticoids, core temperature, activity, neuroinflammation, and circadian gene networks. Female and male Sprague Dawley rats were subjected to clinically relevant thoracic 9 moderate contusion SCI (or laminectomy sham surgery). Diurnal measures-including rhythms of plasma corticosterone (CORT), body temperature, and activity (using small implanted transmitters), and intraspinal circadian and inflammatory gene expression-were studied prior to and after surgery. SCI caused overall increases and disrupted rhythms of the major rodent glucocorticoid, CORT. Presurgery and sham rats displayed expected rhythms in body temperature and activity, whereas rats with SCI had blunted daily rhythms in body temperature and activity. In parallel, SCI disrupted intraspinal rhythms of circadian clock gene expression. Circadian clock genes can act as transcriptional regulators of inflammatory pathways. Indeed, SCI rats also showed dysregulated rhythms in inflammatory gene expression in both the epicenter and distal spinal cord. Our data show that moderate SCI in rats causes wide-ranging diurnal rhythm dysfunction, which is severe at acute time points and gradually recovers over time. Normalizing post-SCI diurnal rhythms could enhance the recovery of homeostasis and quality of life.

Agomelatine treatment corrects symptoms of depression and anxiety by restoring the disrupted melatonin circadian rhythms of rats exposed to chronic constant light

Desynchronization of circadian rhythms is a hallmark of depression. The antidepressant agomelatine, which is an MT1/MT2 melatonin receptor agonist/5-HT2C serotonin receptor antagonist has advantages compared to the selective serotonin reuptake inhibitors as a circadian phase-shifting agent. The present study was designed to explore whether agomelatine is able to have an antidepressant effect on rats exposed to chronic constant light (CCL) for 6 weeks. Focus is also placed on whether this activity affects diurnal rhythms of depressive-like symptoms and is associated with restoration of impaired circadian rhythms in plasma melatonin and corticosterone. We report that CCL induced a depressive-like symptoms associated with decreased grooming in the splash test during the subjective light/inactive phase. Anhedonia-like deficit in the saccharine preference test and increased immobility in the forced swimming test were both detected during the subjective dark/active phase. The disturbed emotional fluctuations due to CCL were corrected by agomelatine treatment (40 mg/kg, i.p. for 3 weeks). Agomelatine also restored novelty-induced hypophagia, which reflects an anxiety state, during the subjective Light and Dark phase, respectively, in rats exposed to CCL. Parallel to the observed positive influence on behavior, this melatonin analogue restored impaired circadian patterns of plasma melatonin but not that of corticosterone. These findings demonstrated the antidepressant-like effect of agomelatine in rats exposed to CCL possibly exerted via correction of melatonin rhythms and are suggestive of the therapeutic potential of this drug in a subpopulation of people characterized by a melatonin deficit.

Effects of L-serine supplementation on the daily rhythms of growth hormone and corticosterone concentrations in mice

The impact of L-serine on the daily rhythms of growth hormone (GH) and corticosterone remains unknown. We explored whether the daily rhythms of these hormones were affected by L-serine supplementation as well as the supplementation time. The results showed that plasma GH concentration at Zeitgeber time (ZT)4 and 8 were significantly increased by L-serine supplementation at ZT22, while the diurnal rhythms peaks of plasma corticosterone at ZT12 were suppressed by L-serine supplementation at ZT10. After the supplementation was stopped, the effects of L-serine on the diurnal rhythms of plasma GH and corticosterone lasted for 2 days then they were fading on day 4. L-serine concentrations in plasma and hypothalamus after supplementation at ZT22 was lower than those after supplementation at ZT10. In conclusion, L-serine modulates the daily rhythms of GH and corticosterone depending on its supplementation time. The modulation effect might be association with the daily rhythms of L-serine metabolism.

Postnatal Ontogeny of the Circadian Expression of the Adrenal Clock Genes and Corticosterone Rhythm in Male Rats.

The postnatal synchronization of the circadian variation of the adrenal clock genes in mammals remains unknown. We evaluated the postnatal ontogeny of daily variation of clock genes (Clock/Bmal1/Per1/Per2/Per3/Cry1/Cry2/Rorα/Rev-Erbα) and steroidogenesis-related genes (Star and Mc2r) in rat adrenals and its relationship with the emergence of plasma corticosterone rhythm using cosinor analysis. Plasma corticosterone circadian rhythm was detected from postnatal day (P)1, with morning acrophase, between zeitgeber time (ZT)0 and ZT2. From P14, there was a nocturnal acrophase of corticosterone at ZT20, which was associated with pups' eye opening. From P3 there was a circadian variation of the mRNA expression of Bmal1, Per2, Per3, and Cry1 genes with morning acrophase, whereas Rev-Erbα had nocturnal acrophase. From P14, Bmal1, Per2, Per3, and Cry1 acrophases advanced by approximately 10 hours, as compared with early neonatal days, becoming vespertine-nocturnal. In all postnatal ages, Per2 and Cry1 circadian profiles were synchronized in phase with the circadian rhythm of plasma corticosterone, whereas Bmal1 was in antiphase. An adult-like Star circadian rhythm profile was observed only from P21. In conclusion, our original data demonstrated a progressive postnatal maturation of the circadian variation of the adrenal clock genes in synchrony with the development of the corticosterone circadian rhythm in rats.

健康と生活習慣病予防における時間栄養学の役割

Almost all metabolic activities of nutrients are under control of the endocrine system, and among them the adrenocortical activity is the most typical example that shows circadian rhythmic changes. We examined relationships between the plasma corticosterone rhythm and feeding schedules in rats. The plasma corticosterone rhythm corresponding to feeding time appears regardless of whether the rats were kept on a light-dark cycle, or in constant light or were blinded. Therefore, it can be concluded that food, more than illumination, is a potent trigger of the circadian adrenocortical rhythm in rats. In addition to the experiments in rats, we also investigated roles of feeding schedulesinformation of the adrenocortical rhythm in human subjects. The corticosterone rhythm disappeared when rats were given a liquid diet intravenously, but not orally, suggesting a necessary role of the oro-gastro-intestinal tract. In addition to the studies in rats, influence to feeding schedules on adrenocortical rhythms was also examined in hospitalized human subjects under a normal light-dark cycle. A clear plasma cortisol rhythm was observed in patients who had been fed on a liquid diet into-duodenum during a restricted time of day, suggest that the circadian cortisol rhythm in man is more closely related to feeding rhythms than cycles of light-dark and sleep-wakefulness. As far as we know, this is the first report demonstrating the important of food in entraining human circadian cortisol rhythm. Next, we measured daily rhythm ofbody temperature in hospitalized patients under continuous and cyclic TEN（totalenteral nutrition）. In the diurnal TEN group, body temperature was low at night and early morning and high in the afternoon early evening. In the continuous TEN group, however, body temperature did not show any significant change with clock-time. In conclusion, the timing of diet intake remarkably modifies the circadian rhythm of body temperature and adrenocortical activity in man. We examined relationship between the feeding time of high salt diet intake and urinary excretion of sodium and chloride. The urinary excretion of sodium and chloride were higher in evening than morning and daytime in woman, is more closely related to the circadian plasma aldosterone rhythm. Furthermore, the clock time difference in endocrine and metabolic responses after physical exercise was assessed according to biorhythm. Physical capacity changed to the time of evaluation, and was greater in the evening than in the morning. Among physiological functions that determine was development of physical strength, plasma growth hormone was higher evening exercise than morning exercise. Further studies are needed on this intriguing idea.

Altered entrainment to the day/night cycle attenuates the daily rise in circulating corticosterone in the mouse.

The suprachiasmatic nucleus (SCN) is a circadian oscillator entrained to the day/night cycle via input from the retina. Serotonin (5-HT) afferents to the SCN modulate retinal signals via activation of 5-HT1B receptors, decreasing responsiveness to light. Consequently, 5-HT1B receptor knockout (KO) mice entrain to the day/night cycle with delayed activity onsets. Since circulating corticosterone levels exhibit a robust daily rhythm peaking around activity onset, we asked whether delayed entrainment of activity onsets affects rhythmic corticosterone secretion. Wheel-running activity and plasma corticosterone were monitored in mice housed under several different lighting regimens. Both duration of the light∶dark cycle (T cycle) and the duration of light within that cycle was altered. 5-HT1B KO mice that entrained to a 9.5L:13.5D (short day in a T = 23 h) cycle with activity onsets delayed more than 4 h after light offset exhibited a corticosterone rhythm in phase with activity rhythms but reduced 50% in amplitude compared to animals that initiated daily activity <4 h after light offset. Wild type mice in 8L:14D (short day in a T = 22 h) conditions with highly delayed activity onsets also exhibited a 50% reduction in peak plasma corticosterone levels. Exogenous adrenocorticotropin (ACTH) stimulation in animals exhibiting highly delayed entrainment suggested that the endogenous rhythm of adrenal responsiveness to ACTH remained aligned with SCN-driven behavioral activity. Circadian clock gene expression in the adrenal cortex of these same animals suggested that the adrenal circadian clock was also aligned with SCN-driven behavior. Under T cycles <24 h, altered circadian entrainment to short day (winter-like) conditions, manifest as long delays in activity onset after light offset, severely reduces the amplitude of the diurnal rhythm of plasma corticosterone. Such a pronounced reduction in the glucocorticoid rhythm may alter rhythmic gene expression in the central nervous system and in peripheral organs contributing to an array of potential pathophysiologies.

Acute inhibition of casein kinase 1δ/ε rapidly delays peripheral clock gene rhythms.

Circadian rhythms are generated through a transcription-translation feedback loop involving clock genes and the casein kinases CSNK1D and CSNK1E. In this study, we investigated the effects of the casein kinase inhibitor PF-670462 (50 mg/kg) on rhythmic expression of clock genes in the liver, pancreas and suprachiasmatic nucleus (SCN) as well as plasma corticosterone, melatonin and running behaviour in rats and compared them to the responses to a 4 h extension of the light phase. PF-670462 acutely phase delayed the rhythmic transcription of Bmal1, Per1, Per2 and Nr1d1 in both liver and pancreas by 4.5 ± 1.3 and 4.5 ± 1.2 h, respectively, 1 day after administration. In the SCN, the rhythm of Nr1d1 and Dbp mRNA expression was delayed by 4.2 and 4 h, respectively. Despite these changes, the time of peak plasma melatonin secretion was not delayed, although the plasma corticosterone rhythm and onset of wheel-running activity were delayed by 2.1 and 1.1 h, respectively. These changes are in contrast to the effects of the 4 h light extension, which resulted in delays in peak expression of the clock genes of less than 1 h and no change in the melatonin or corticosterone rhythms. The ability of the casein kinase inhibitor to bring about large phase shifts in the rhythms of major metabolic target tissues may lead to new drugs being developed to rapidly phase adjust circadian rhythms to alleviate the metabolic impact of shift work.

Programming of the Hypothalamic-Pituitary-Adrenal Axis by Neonatal Intermittent Hypoxia: Effects on Adult Male ACTH and Corticosterone Responses Are Stress Specific

Intermittent hypoxia (IH) is an animal model of apnea-induced hypoxia, a common stressor in the premature neonate. Neonatal stressors may have long-term programming effects in the adult. We hypothesized that neonatal exposure to IH leads to significant changes in basal and stress-induced hypothalamic-pituitary-adrenal (HPA) axis function in the adult male rat. Rat pups were exposed to normoxia (control) or 6 approximately 30-second cycles of IH (5% or 10% inspired O₂) daily on postnatal days 2-6. At approximately 100 days of age, we assessed the diurnal rhythm of plasma corticosterone and stress-induced plasma ACTH and corticosterone responses, as well as mRNA expression of pertinent genes within the HPA axis. Basal diurnal rhythm of plasma corticosterone concentrations in the adult rat were not affected by prior exposure to neonatal IH. Adults exposed to 10% IH as neonates exhibited an augmented peak ACTH response and a prolonged corticosterone response to restraint stress; however, HPA axis responses to insulin-induced hypoglycemia were not augmented in adults exposed to neonatal IH. Pituitary Pomc, Crhr1, Nr3c1, Nr3c2, Avpr1b, and Hif1a mRNA expression was decreased in adults exposed to neonatal 10% IH. Expression of pertinent hypothalamic and adrenal mRNAs was not affected by neonatal IH. We conclude that exposure to neonatal 10% IH programs the adult HPA axis to hyperrespond to acute stimuli in a stressor-specific manner.

Forced Desynchrony Reveals Independent Contributions of Suprachiasmatic Oscillators to the Daily Plasma Corticosterone Rhythm in Male Rats

The suprachiasmatic nucleus (SCN) is required for the daily rhythm of plasma glucocorticoids; however, the independent contributions from oscillators within the different subregions of the SCN to the glucocorticoid rhythm remain unclear. Here, we use genetically and neurologically intact, forced desynchronized rats to test the hypothesis that the daily rhythm of the glucocorticoid, corticosterone, is regulated by both light responsive and light-dissociated circadian oscillators in the ventrolateral (vl-) and dorsomedial (dm-) SCN, respectively. We show that when the vlSCN and dmSCN are in maximum phase misalignment, the peak of the plasma corticosterone rhythm is shifted and the amplitude reduced; whereas, the peak of the plasma adrenocorticotropic hormone (ACTH) rhythm is also reduced, the phase is dissociated from that of the corticosterone rhythm. These data support previous studies suggesting an ACTH-independent pathway contributes to the corticosterone rhythm. To determine if either SCN subregion independently regulates corticosterone through the sympathetic nervous system, we compared unilateral adrenalectomized, desynchronized rats that had undergone either transection of the thoracic splanchnic nerve or sham transection to the remaining adrenal. Splanchnicectomy reduced and phase advanced the peak of both the corticosterone and ACTH rhythms. These data suggest that both the vlSCN and dmSCN contribute to the corticosterone rhythm by both reducing plasma ACTH and differentially regulating plasma corticosterone through an ACTH- and sympathetic nervous system-independent pathway.

Impaired release of corticosterone from adrenals contributes to impairment of circadian rhythms of activity in hyperammonemic rats

Patients with liver cirrhosis may present impaired sleep-wake and circadian rhythms, relative adrenal insufficiency and altered hypothalamus–pituitary–adrenal gland (HPA) axis. The underlying mechanisms remain unclear. Circadian rhythms are modulated by corticosteroids which secretion is regulated by HPA axis. Hyperammonemia alters circadian rhythms of activity and corticosterone in rats.The aims were: (1) assessing whether corticosterone alterations are responsible for altered circadian rhythm in hyperammonemia: (2) to shed light on the mechanism by which corticosterone circadian rhythm is altered in hyperammonemia.The effects of daily corticosterone injection at ZT10 on circadian rhythms of activity, plasma corticosterone, adreno-corticotropic hormone (ACTH) and hypothalamic corticotropic releasing hormone (CRH) were assessed in control and hyperammonemic rats. ACTH-induced corticosterone release was analyzed in cultured adrenal cells.Corticosterone injection restores the corticosterone peak in hyperammonemic rats and their activity and circadian rhythm. Plasma ACTH and CRH in hypothalamus are increased in hyperammonemic rats. Corticosterone injection normalizes ACTH.Chronic hyperammonemia impairs adrenal function, reduces corticosterone content and ACTH-induced corticosterone release in adrenals, leading to reduced feedback modulation of HPA axis by corticosterone which contributes to impair circadian rhythms of activity. Impaired circadian rhythms and motor activity may be corrected in hyperammonemia and hepatic encephalopathy by corticosterone treatment.

Photoperiodic lighting (16 hours of light:8 hours of dark) programs during incubation: 1. Effects on growth and circadian physiological traits of embryos and early stress response of broiler chickens

This study was conducted to evaluate the effect of a 16L:8D photoperiod during incubation, either during the whole incubation period (Inc(0-21d)) or the last week of incubation (Inc(14-21d)), on embryo growth, incubation performance, and light:dark rhythm of plasma melatonin and corticosterone in relation to early stress responses of newly hatched chicks to the posthatching environment. A dark incubation condition (Inc(Dark)) served as control. Three batches of eggs (n = 1,080, 1,320, 720) from Ross 308 broiler breeders were used in the experiment. Embryos from Inc(0-21d) presented a daily rhythm of melatonin at internal pipping and hatching, but Inc(Dark) embryos did not. The Inc(14-21d) group had rhythmic plasma melatonin at hatching only. A L:D rhythm of corticosterone was apparent at hatching. A significant incubation × sampling time interaction suggested that a lower increment in blood corticosterone level in Inc(0-21d) at 8 h posthatching (light period), as compared with hatching (dark period) values, might be associated with probable changes in the hypothalamic-pituitary-adrenal axis in Inc(0-21d) through incubation lighting. This finding may also suggest improved adaptation to the posthatching environment. Incubation lighting did not consistently affect brain malondialdehyde concentration; the only difference between groups was higher concentrations at hatching in Inc(14-21d), whereas incubation groups at the internal pipping stage had similar values. Mean relative asymmetry (RA) did not differ with incubation lighting. The malondialdehyde and RA results indicate that neither lighting nor darkness during the overall incubation exacerbated embryo oxidative and developmental stress. An increased breast muscle weight was observed at hatching only in Inc(14-21d). The Inc(0-21d) group had increased embryo weights relative to egg weight and decreased residual yolk but had no effect on chick weight, relative heart and liver (% of embryo weight), hatchability, embryo mortality, incubation time, oxidative stress, or mean RA. In conclusion, these results provide further evidence that photoperiodic lighting during incubation (Inc(0-21d)) may improve adaptation of chicks to a novel environment at hatching, possibly giving birds a better start for early posthatching development.

Setting the main circadian clock of a diurnal mammal by hypocaloric feeding

Caloric restriction attenuates the onset of a number of pathologies related to ageing. In mammals, circadian rhythms, controlled by the hypothalamic suprachiasmatic (SCN) clock, are altered with ageing. Although light is the main synchronizer for the clock, a daily hypocaloric feeding (HF) may also modulate the SCN activity in nocturnal rodents. Here we report that a HF also affects behavioural, physiological and molecular circadian rhythms of the diurnal rodent Arvicanthis ansorgei. Under constant darkness HF, but not normocaloric feeding (NF), entrains circadian behaviour. Under a light–dark cycle, HF at midnight led to phase delays of the rhythms of locomotor activity and plasma corticosterone. Furthermore, Per2 and vasopressin gene oscillations in the SCN were phase delayed in HF Arvicanthis compared with animals fed ad libitum. Moreover, light-induced expression of Per genes in the SCN was modified in HF Arvicanthis, despite a non-significant effect on light-induced behavioural phase delays. Together, our data show that HF affects the circadian system of the diurnal rodent Arvicanthis ansorgei differentially from nocturnal rodents. The Arvicanthis model has relevance for the potential use of HF to manipulate circadian rhythms in diurnal species including humans.

Variable Restricted Feeding Disrupts the Daily Oscillations of Period2 Expression in the Limbic Forebrain and Dorsal Striatum in Rats

Predictable restricted feeding schedules limit food availability to a single meal at the same time each day, lead to the induction and entrainment of circadian rhythms in food-anticipatory activity, and shift daily rhythms of clock gene expression in areas of the brain that are important in the regulation of motivational and emotional state. In contrast, when food is delivered under a variable restricted feeding (VRF) schedule, at a new and unpredictable mealtime each day, circadian rhythms in food-anticipatory activity fail to develop. Here, we study the effects of VRF on the daily rhythm of plasma corticosterone and of clock gene expression in the limbic forebrain and dorsal striatum, of rats provided a 2-h access to a complete meal replacement (Ensure Plus) at an unpredictable time each day. VRF schedules varied the mealtimes within the 12h of light (daytime VRF), the 12h of dark (nighttime VRF), or across the 24h light-dark cycle (anytime VRF). Our results show that contrary to the synchronizing effects of predictable restricted feeding, VRF blunts the daily corticosterone rhythm and disrupts daily rhythms of PER2 expression in a region-specific and mealtime-dependent manner.

Sex and Dosing-Time Dependencies in Irinotecan-Induced Circadian Disruption

Circadian disruption accelerates malignant growth; thus, it should be avoided in anticancer therapy. The circadian disruptive effects of irinotecan, a topoisomerase I inhibitor, was investigated according to dosing time and sex. In previous work, irinotecan achieved best tolerability following dosing at zeitgeber time (ZT) 11 in male and ZT15 in female mice, whereas worst toxicity corresponded to treatment at ZT23 and ZT3 in male and female mice, respectively. Here, irinotecan (50 mg/kg intravenous [i.v.]) was delivered at the sex-specific optimal or worst circadian timing in male and female B6D2F1 mice. Circadian disruption was assessed with rest-activity, body temperature, plasma corticosterone, and liver mRNA expressions of clock genes Rev-erbα, Per2, and Bmal1. Baseline circadian rhythms in rest-activity, body temperature, and plasma corticosterone were more prominent in females as compared to males. Severe circadian disruption was documented for all physiology and molecular clock endpoints in female mice treated at the ZT of worst tolerability. Conversely, irinotecan administration at the ZT of best tolerability induced slight alteration of circadian physiology and clock-gene expression patterns in female mice. In male mice, irinotecan produced moderate alterations of circadian physiology and clock-gene expression patterns, irrespective of treatment ZT. However, the average expression of Rev-erbα, Per2, and Bmal1 were down-regulated 2- to 10-fold with irinotecan at the worst ZT, while being minimally or unaffected at the best ZT, irrespective of sex. Corticosterone secretion increased acutely within 2 h with a sex-specific response pattern, resulting in a ZT-dependent phase-advance or -delay in both sex. The mRNA expressions of irinotecan clock-controlled metabolism genes Ce2, Ugt1a1, and Top1 were unchanged or down-regulated according to irinotecan timing and sex. This study shows that the circadian timing system represents an important toxicity target of irinotecan in female mice, where circadian disruption persists after wrongly timed treatment. As a result, the mechanisms underling cancer chronotherapeutics are expectedly more susceptible to disruption in females as compared to males. Thus, the optimal circadian timing of chemotherapy requires precise determination according to sex, and should involve the noninvasive monitoring of circadian biomarkers. (Author correspondence: francis.levi@inserm.fr)

Rhythmic expression of per1 in the dentate gyrus is suppressed by corticosterone: Implications for neurogenesis

The stimulating action of anti-depressant drugs on the number of mitotic cells on the dentate gyrus on the adult rat depends on an intact diurnal rhythm of corticosterone. This suggests that there may be a clock mechanism in the dentate gyrus which is sensitive to corticoids. This paper reports the diurnal expression the ‘clock’ gene per1 in the dentate gyrus, and how it is altered by clamping the diurnal rhythm in corticosterone. We show that there is a diurnal rhythm in the number of mitotic progenitor cells in the dentate gyrus of the hippocampus in adult male per1-luciferase rats, approximately 6 h out of phase with the plasma corticosterone rhythm. This is suppressed by clamping the daily corticosterone levels by a subcutaneous implant of corticosterone (100 mg). There was also a daily rhythm of per1 expression in both suprachiasmatic nucleus (SCN) and the dentate gyrus, which were in phase with one another. The per1 rhythm in the dentate gyrus, but not the SCN, was suppressed by clamping the plasma corticosterone rhythm. These results are related to the previous finding that clamping the corticosterone rhythm also prevents the stimulating action of fluoxetine and other controlling agents on the mitotic activity of the progenitor cells.

Depression‐like behavior in the forced swimming test in PACAP‐deficient mice: amelioration by the atypical antipsychotic risperidone

Pituitary adenylate cyclase-activating polypeptide (PACAP) is a neuropeptide with pleiotropic functions. We report here that PACAP-deficient (PACAP-/-) mice showed increased immobility in a forced swimming test, which was reduced by the antidepressant desipramine, to a similar extent as in wild-type mice. The atypical antipsychotic risperidone and the selective serotonin (5-HT)(2) antagonist ritanserin normalized the depression-like behavior in PACAP-/- mice. The 5-HT(2) agonist (+/-)-2,5-dimethoxy-4-iodoamphetamine-induced 5-HT syndrome was exaggerated in PACAP-/- mice, which suggests a 5-HT(2)-receptor-dependent mechanism in the depression-like behavior. The circadian rhythm of plasma corticosterone and body core temperature was significantly flattened in the mutants. mRNA expression of glucocorticoid receptor was reduced in the mutant hippocampus. The present results suggest that alterations in PACAP signaling might contribute to the pathogenesis of certain depressive conditions amenable to atypical antipsychotic drugs.

Diurnal expression of period 2 and urocortin 1 in neurones of the non-preganglionic Edinger-Westphal nucleus in the rat

Period 2 (Per2) is an important clock gene involved in the regulation of the major circadian clock in the mammalian central nervous system, the suprachiasmatic nucleus. In addition, Per2 is expressed in many other stress-sensitive brain structures. We have previously showed that the non-preganglionic Edinger-Westphal nucleus (npEW) is the main site of the corticotropin-releasing factor peptide family member urocortin 1 (Ucn1) and that this peptide undergoes conspicuous expression changes in response to various stressors. Here, we hypothesized that in the rat npEW both Per2 and Ucn1 would be produced in a diurnal, rhythmical fashion. This hypothesis was tested by following this expected rhythm on two days in rats killed at four time points each day (Zeitgeber times 0, 6, 12, and 18). We showed the co-existence of Per2 and Ucn1 in the npEW with double-label immunofluorescence and demonstrated with quantitative RT-PCR and semi-quantitative immunocytochemistry diurnal rhythms in Per2 mRNA expression and Per2 protein content, each on a single different day, with a minimum at lights-off and a maximum at lights-on. We furthermore revealed a diurnal rhythm in the number of Ucn1-immunopositive neurones and in their Ucn1 peptide content, with a minimum at night and at the beginning of the light period and a peak at lights-off, while the Ucn1 mRNA content paralleled the Per2 mRNA rhythm. The rhythms were accompanied by a diurnal rhythm in plasma corticosterone concentration.Our results are in line with the hypothesis that both Per2 and Ucn1 in the rat npEW are produced in a diurnal fashion, a phenomenon that may be relevant for the regulation of the diurnal rhythm in the stress response.