Evidence that GAD65 mediates increased GABA synthesis during intense neuronal activity in vivo

In this study we tested the hypothesis that the 65-kDa isoform of glutamate decarboxylase (GAD(65)) mediates activity-dependent GABA synthesis as invoked by seizures in anesthetized rats. GABA synthesis was measured following acute GABA-transaminase inhibition by gabaculine using spatially localized (1)H NMR spectroscopy before and after bicuculline-induced seizures. Experiments were conducted with animals pre-treated with vigabatrin 24 h earlier in order to reduce GAD(67) protein and also with non-treated controls. GAD isoform content was quantified by immunoblotting. GABA was higher in vigabatrin-treated rats compared to non-treated controls. In vigabatrin-treated animals, GABA synthesis was 28% lower compared to controls [p < 0.05; vigabatrin-treated, 0.043 +/- 0.011 micromol/(g min); non-treated, 0.060 +/- 0.014 micromol/(g min)] and GAD(67) was 60% lower. No difference between groups was observed for GAD(65). Seizures increased GABA synthesis in both control [174%; control, 0.060 +/- 0.014 micromol/(g min) vs. seizures, 0.105 +/- 0.043 micromol/(g min)] and vigabatrin-treated rats [214%; control, 0.043 +/- 0.011 micromol/(g min); seizures, 0.092 +/- 0.018 micromol/(g min)]. GAD(67) could account for at least half of basal GABA synthesis but only 20% of the two-fold increase observed in vigabatrin-treated rats during seizures. The seizure-induced activation of GAD(65) in control cortex occurs concomitantly with a 2.3-fold increase in inorganic phosphate, known to be a potent activator of apoGAD(65)in vitro. Our results are consistent with a major role for GAD(65) in activity-dependent GABA synthesis.     

Time-dependent Etomidate-induced Anesthesia in Hamsters

In this study, we assessed the temporal variation in the hypnotic effect of etomidate, a drug that positively modulates GABA A -mediated neurotransmission in the Syrian hamster. Ten to twenty mg/kg of etomidate i.p. induced a significant loss of righting reflex when administered at 12:00 h but not 24:00 h in hamsters housed under a 14 : 10 light : dark photoperiod (lights on at 06:00 h). On the contrary, ketamine (administered together with xylazine), which induces anesthesia by inhibiting glutamatergic neurotransmission, did not show a diurnal variation in the loss of righting reflex test. These results support previous data regarding diurnal rhythms in GABA content and activity. The phase dependent response to etomidate may be due to daily allosteric modulation of the GABA_A receptor.     

Circadian variation of carrageenan-paw edema in the rat

The circadian variation of carrageenan (carr)-induced paw edema was studied in sham-operated and adrenalectomized rats. The edema was produced by carr injection into the plantar tissue at 03:00, 09:00, 15:00 and 20:00 hr. In sham-operated rats the rate of appearance of maximal edema was much faster in the evening than in the morning: at 20:00 hr, it was obtained 2 hours after carr injection while at 09:00 hr it took 4 hours. At 03:00 and 15:00 hr, maximal edema was found respectively 2.5 and 4 hours after carr. In adrenalectomized rats, maximal edema at 4 hours of investigation was always larger than in sham-operated animals but the rate of appearance of edema did not change throughout the day as it was obtained 3 hours after carr administration. At 09:00 and 20:00 hr injection of hydrocortisone (HC) to adrenalectomized rats produce the same dose-dependent effect on the rate of formation of edema. However, to reproduce in adrenalectomized animals the rates of formation of edema similar to those obtained in sham-operated rats, an injection of 18 mg/kg HC was required at 09:00 hr while less than 2 mg/kg was needed at 20:00 hr. The results suggest that the circadian rhythm of carr edema is related to circadian variation in the corticosteroid system.     

Time-of-day effect on the torque and neuromuscular properties of dominant and non-dominant quadriceps femoris

The study was conducted first, to determine the possibility of a dichotomy between circadian rhythm of maximal torque production of the knee extensors of the dominant and non-dominant legs, and second, to determine whether the possible dichotomy could be linked to a change in the downward drive of the central nervous system and/or to phenomena prevailing at the muscular level. The dominant leg was defined as the one with which subjects spontaneously kick a football. Tests were performed at 06:00, 10:00, 14:00, 18:00, and 22:00 h. To distinguish the neural and muscular mechanisms that influence muscle strength, the electromyographic and mechanical muscle responses associated with electrically evoked and/or voluntary contractions of the human quadriceps and semi-tendinosus muscles for each leg were recorded and compared. The main finding was an absence of interaction between time-of-day and dominance effects on the torque associated with maximal voluntary contraction (MVC) of both quadriceps. A significant time-of-day effect on MVC torque of the knee extensors was observed for the dominant and non-dominant legs when the data were collapsed, with highest values occurring at 18:00 h (p < 0.01). From cosinor analysis, a circadian rhythm was documented (p < 0.001) with the peak (acrophase) estimated at 18:18 +/- 00:12 h and amplitude (one-half the peak-to-trough variation) of 3.3 +/- 1.1%. Independent of the leg tested, peripheral mechanisms demonstrated a significant time-of-day effect (p < 0.05) on the peak-torque of the single and doublet stimulations, with maximal levels attained at 18:00 h. The central activation of the quadriceps muscle of each leg remained unchanged during the day. The present results confirmed previous observations that muscle torque changes in a predictable manner during the 24 h period, and that the changes are linked to modifications prevailing at the muscular, rather than the neural, level. The similar rhythmicity observed in this study between the dominant and non-dominant legs provides evidence that it is not essential to test both legs when simple motor tasks are investigated as a function of the time of day.     

Circadian Changes in Anxiolysis-Related Behavior of Syrian Hamsters. Correlation with Hypothalamic GABA Release

The diurnal variations in anxiolysis and exploratory behavior were examined in a plus-maze paradigm in Syrian hamsters exposed to 14 h light: 10 h dark photoperiods or to constant darkness for 3 days. The percent of time spent in open arms and the percent of entries to open arms (both indexes of anxiety-related behavior) as well as the number of crosses to both arms (an index of locomotor behavior) showed significant daily variations under the two lighting conditions, maxima being found at night (2400-0400 h). Flumazenil (5 mg/kg) injected at the middle of the light (at 1600 h) or dark period (at 0400 h) decreased by 39-54 % anxiolysis-related behavior without affecting locomotor activity significantly. 3 H-?-aminobutyric acid (GABA) release from preoptic-medial basal hypothalamic explants obtained from hamsters exposed to 14 h light: 10 h dark photoperiods attained maximal values at 2400-0400 h. The results further supported the existence of circadian changes in anxiolysis-related behavior in Syrian hamsters that correlated with an increased hypothalamic GABA release.     

Stimulation of Synaptosomal γ-Aminobutyric Acid Synthesis by Glutamate and Glutamine

gamma-Aminobutyric acid (GABA) synthesis was studied in rat brain synaptosomes by measuring the increase of GABA level in the presence of the GABA-transaminase inhibitor gabaculine. The basal rate of synaptosomal GABA synthesis in glucose-containing medium (25.9 nmol/h/mg of protein) was only 3% of the maximal activity of glutamate decarboxylase (GAD; 804 +/- 83 nmol/h/mg of protein), a result indicating that synaptosomal GAD operates at only a small fraction of its catalytic capacity. Synaptosomal GABA synthesis was stimulated more than threefold by adding 500 microM glutamine. Glutamate also stimulated GABA synthesis, but the effect was smaller (1.5-fold). These results indicate that synaptosomal GAD is not saturated by endogenous levels of its substrate, glutamate, and account for part of the unused catalytic capacity. The greater stimulation of GABA synthesis by glutamine indicates that the GAD-containing compartment is more accessible to extrasynaptosomal glutamine than glutamate. The strong stimulation by glutamine also shows that the rates of uptake of glutamine and its conversion to glutamate can be sufficiently rapid to support GABA synthesis in nerve terminals. Synaptosomes carried out a slow net synthesis of aspartate in glucose-containing medium (7.7 nmol/h/mg of protein). Aspartate synthesis was strongly stimulated by glutamate and glutamine, but in this case the stimulation by glutamate was greater. Thus, the larger part of synaptosomal aspartate synthesis occurs in a different compartment than does GABA synthesis.     

Circadian rhythm of airways caliber and its autonomic modulation

ABSTRACT

The autonomic nervous system (ANS) is one of the effector pathways for circadian variation of many physiological parameters. Autonomic tone and airways caliber have been reported to exhibit circadian variation in separate studies. A simultaneous investigation of heart rate variability (HRV) and airway caliber might ascertain how airway caliber is modulated by autonomic tone. This study was planned to identify the variations in airway caliber and autonomic function tone during a 24-hour span. A total of 56 healthy male subjects with almost similar daily routines were studied. Time domain, frequency domain and nonlinear analysis of R-R interval from 5 min electrocardiogram (ECG) was done seven times during the daytime wake span at 3-hour intervals starting at 05:00 h in the morning until 23:00 h in the night. Simultaneously peak expiratory flow rate (PEFR) was determined using a mini Wright’s peak flow meter. Rhythmometric analysis was done for PEFR and HRV parameters. Significant circadian variation in low frequency (LF) and high frequency (HF) variance was identified in this group of healthy subjects. The circadian rhythm of LF variance was characterized by a gradual increase and corresponding reciprocal change in HF variance from morning until night. The LF/HF ratio and SD2/SD1 ratio reflecting sympatho-vagal balance showed low to high values from morning to evening. The acrophase of the PEFR temporal pattern is similar to that of LF power and almost opposite in phase to that of HF power. PEFR is positively correlated with LF power. The circadian rhythm of airway caliber co-varies with cardiac autonomic tone. It appears that the temporal pattern of cardiac autonomic tone precedes in time that of airways caliber, thereby suggesting the latter operates under the modulatory effect of the 24-hour pattern in sympatho-vagal balance.     

中缅树鼩体温、代谢率和蒸发失水日节律

为研究中缅树鼩(Tupaia belangeri)体温、代谢率和蒸发失水的日节律变化,采用植入式体温计测定了中缅树鼩24 h的体温,以及24 h中4个时间段(05:00～07:00时、11:00～13:00时、17:00～19:00时和23:00～01:00时)热中性区(30℃)的静止代谢率(RMR)、非颤抖性产热(NST)和蒸发失水(EWL)。结果显示,中缅树鼩的体温具有日节律变化,最高值和最低值分别出现在11:00时和03:00时,各为(39.45±0.26)℃和(36.34±0.24)℃;静止代谢率、非颤抖性产热和蒸发失水在4个时间段都有显著差异,表现出显著的日节律变化,代谢率在23:00～01:00时最大,O2含量为(2.58±0.04)ml/(g.h),在11:00～13:00时最小,O2含量为(2.28±0.09)ml/(g.h);非颤抖性产热在05:00～07:00时最大,O2含量为(3.08±0.14)ml/(g.h),在11:00～13:00最小,O2含量为(2.69±0.63)ml/(g.h);蒸发失水在17:00～19:00时最大,失水量为(3.60±0.31)mg/(g.h)。结果表明,体温的日节律变化主要与环境温度的日节律变化和下午出窝取食活动性增强有关;当夜晚环境温度相对较低的时候,通过增强静止代谢率和非颤抖性产热来增加产热,而白天环境温度相对较高的时候,通过增强蒸发失水散热来调节体温。     

Effect of ACTH (tetracosactide) on steroid hormone levels in the mare. Part B: effect in ovariectomized mares (including estrous behavior)

The mare is the only non-primate species known to display estrous signs after ovariectomy and adrenal hormones have been implicated as a possible cause. Moreover, in several species, estradiol seems to have a stimulatory effect on the hypothalamic-pituitary-adrenal axis. The aim of the present study was to compare the effect of ACTH (tetracosactide) on pertinent hormones [cortisol, progesterone, androstenedione, testosterone (intact and ovariectomized mares) and estradiol (ovariectomized mares only)] in intact mares in estrus with the same mares after ovariectomy (n=5). Blood samples were collected hourly from 12:00 until 14:00 h the following day (half-hourly between 14:00 and 17:00 h) on two occasions, with saline or ACTH treatment at 14:00 h (saline treatment day or ACTH treatment day). The mares, both when intact and after ovariectomy, showed a significant increase in all measured hormones, except estradiol (not measured in intact mares), after ACTH treatment, lasting at least 3h post-treatment (P<0.001). On the saline treatment day, cortisol levels in ovariectomized mares were lower than in intact mares in the evening (18:00-23:00 h), but higher at night (24:00-05:00 h). No differences in cortisol response between mares, when intact and after ovariectomy, were found after ACTH treatment (P=0.3). Androstenedione levels were lower (P<0.001) and increased less after ACTH treatment in ovariectomized mares, as compared to when intact (P<0.05). Progesterone concentrations were lower in the ovariectomized mares at night (24:00-05:00 h) on the saline treatment day and at all times on the ACTH treatment day (P<0.05). Testosterone concentrations were lower in ovariectomized mares on both treatment days, as compared to when intact (P<0.001). It was concluded that ovariectomy affected basal cortisol pattern. Ovarian androstenedione and testosterone contributed to the basal circulating levels and, in the case of androstenedione, was stimulated by ACTH. Endogenous estradiol did not act stimulatory on adrenal gland hormone production in the mare.     

Orcadian Rhythm in the Torque Developed by Elbow Flexors During Isometric Contraction Effect of Sampling Schedules

Time-dependent changes in elbow flexion torque have been documented according to two different sampling schedules. Seven physical education students took part in the first series of experiments, and 7 other similar subjects in the second. In both sets of experiments, the subjects performed isometric contractions: maximal and submaximal at 90° in the first experiments and maximal at different angular positions in the second. After a 30-minute rest period, the torque developed was measured at 00:00, 06:00, 09:00, 12:00, 15:00, 18:00, and 21:00h on the day of the experiment. These subjects remained in the laboratory for 24h. In the second series of experiments, the torque developed was measured at 01:00, 05:00, 09:00, 13:00, 17:00, and 21:00h over the subsequent 6 days with only one test session per day. In this case, there was an interval of 20h between two successive test sessions. In the first experiment, a significant time-of-day effect was observed for the torque of the elbow flexors under isometric conditions with an acrophase at 17:58h. The 24h normalized mean score was 92.85% with an amplitude of 7.63% of the daily mean. In the second series of experiments, there was evidence of a circadian rhythm in the torque developed by the elbow flexors at every angle position, especially at 90°, the angle investigated in the first set of experiments. The peak torque was calculated to have occurred at 17:55h. The amplitude of the rhythm was equal to 6.99% of the daily mean. There were no statistically significant differences in the characteristics of the circadian rhythm observed between the two experimental designs. We concluded that an experiment extending over several days could be employed to evaluate circadian rhythms in muscular activity reliably. (Chronobiology International, 14(3), 287–294, 1997)     

5-methoxypsoralen: Effect on the noradrenaline circadian rhythm of sleep-deprived subjects

5-methoxypsoralen (5-MOP), a serotonin analogue, stimulates the secretion of melatonin, which plays an important role in circadian rhythm regulation. Melatonin production is essentially controlled by noradrenaline (NA). To investigate the effect of 5-MOP on the 24 hr NA rhythm, hourly plasma NA concentrations measured over 24 hr in 7 healthy young subjects who took 40 mg 5-MOP orally at 21:00 hr the evening before were compared with values obtained in drug-free subjects. All subjects were sleep-deprived and under conditions in which sympathetic nervous system activation was repeated every hour over 24 hr. In both series of patients, a significant difference was observed between mean morning values and mean night values, reflecting persistence of a circadian rhythm. In subjects given 5-MOP, a significant difference was also observed between mean afternoon values and mean night values. However, MANOVA analysis failed to find any difference between the two series of subjects. Acute administration of 5-MOP thus had no significant effect on the NA circadian rhythm under our study conditions.     

Daily changes of GABA and taurine concentrations in various hypothalamic areas are affected by chronic hyperprolactinemia

This study was designed to characterize, in anterior, mediobasal, and posterior hypothalamic and median eminence, the 24h changes of gamma aminobutyric acid (GABA) and taurine (TAU) contents in adult male rats and to analyze whether chronic hyperprolactinemia may affect these patterns. Rats were turned hyperprolactinemic by a pituitary graft. Plasma prolactin (PRL) levels increased after pituitary grafting at all time points examined. A disruption of the circadian rhythm was observed in pituitary-grafted rats, whereas GABA and TAU content followed daily rhythms in all areas studied in controls. In the mediobasal hypothalamus, two peaks for each amino acid were found at midnight and midday. In the anterior hypothalamus, GABA and TAU showed only one peak of concentration at midnight. In the posterior hypothalamus, the values of both GABA and TAU were higher during the light as compared to the dark phase of the photoperiod. In the median eminence GABA content peaked at 20:00h, the time when TAU exhibited the lowest values. Hyperprolactinemia abolished the 24h changes of GABA in the mediobasal hypothalamus and reduced its content as compared to controls. Hyperprolactinemia advanced the diurnal peak of TAU to 12:00h in the mediobasal hypothalamus and did not modify the 24:00h peak. In the anterior hypothalamus, hyperprolactinemia increased GABA and TAU contents during the light phase while it decreased them during the dark phase of the photoperiod. In the posterior hypothalamus hyperprolactinemia did not modify GABA or TAU patterns as compared to controls. In the median eminence hyperprolactinemia increased the 20:00h peak of GABA and shift advanced the decrease in TAU content at 20:00h and its maximum at 24:00h as compared to controls. These data show that GABA and TAU content exhibit specific daily patterns in each hypothalamic region studied. PRL differentially affects the daily pattern of these amino acids in each hypothalamic region analyzed.     

Neuro-endocrine correlations of hypothalamic-pituitary-thyroid axis in healthy humans

Neuro-endocrine hormone secretion is characterized by circadian rhythmicity. Melatonin, GRH and GH are secreted during the night, CRH and ACTH secretion peak in the morning, determining the circadian rhythm of cortisol secretion, TRH and TSH show circadian variations with higher levels at night. Thyroxine levels do not change with clear circadian rhythmicity. In this paper we have considered a possible influence of cortisol and melatonin on hypothalamic-pituitary-thyroid axis function in humans. Melatonin, cortisol, TRH, TSH and FT4 serum levels were determined in blood samples obtained every four hours for 24 hours from ten healthy males, aged 36-51 years. We correlated hormone serum levels at each sampling time and evaluated the presence of circadian rhythmicity of hormone secretion. In the activity phase (06:00 h-10:00 h-14:00 h) cortisol correlated negatively with FT4, TSH correlated positively with TRH, TRH correlated positively with FT4 and melatonin correlated positively with TSH. In the resting phase (18:00 h-22:00 h-02:00 h) TRH correlated positively with FT4, melatonin correlated negatively with FT4, TSH correlated negatively with FT4, cortisol correlated positively with FT4 and TSH correlated positively with TRH. A clear circadian rhythm was validated for the time-qualified changes of melatonin and TSH secretion (with acrophase during the night), for cortisol serum levels (with acrophase in the morning), but not for TRH and FT4 serum level changes. In conclusion, the hypothalamic-pituitary-thyroid axis function may be modulated by cortisol and melatonin serum levels and by their circadian rhythmicity of variation.     

Circadian phase modulates the enhancing effect of the Egyptian Moringa peregrina extract on learning and memory in mice

One of the most important issues nowadays is memory disorders. The exact circadian modulation of learning and memory is still under investigation. The present study was carried out to evaluate the probable enhancing and neuroprotective effects of the Egyptian Moringa peregrina; administered at two different Zeitgeber times (ZT), on learning and memory in mice. M. peregrina dried leaves were aqueously extracted and intranasally (IN) administered at four doses (viz. 12.5, 25, 50, and 100 mg/kg). Twenty animal groups (n = 8/group) were treated with M. peregrina at ZT 3:00 vs. 15:00 representing the rest (day) vs. active (night) phases, respectively. Additionally, sub-chronic effect of daily IN administration of the 25 mg/kg dose for two weeks was examined at ZT 3:00 vs. 15:00. Memory performance was assessed after 1, 2 weeks of treatment and a 3rd recovery week. Memory test was performed at ZT 17:00 by measuring the percentage of time spent in novel arm of Y-maze. Dose-response curve revealed that both 25 and 50 mg/kg doses administered at ZT 3:00 significantly improved memory efficiency. Rest phase administration significantly enhanced memory functions after 1, 2 weeks as well as after the recovery week. Moreover, IN pretreatment with M. peregrina (25 mg/kg) revealed a protective effect against scopolamine-induced amnesia. In conclusion, these findings clearly indicated that M. peregrina possesses valuable enhancing and protective effects on learning and memory processes in mice which is circadian-phase dependent.     

Daily Changes of GABA and Taurine Concentrations in Various Hypothalamic Areas Are Affected by Chronic Hyperprolactinemia

This study was designed to characterize, in anterior, mediobasal, and posterior hypothalamic and median eminence, the 24h changes of gamma aminobutyric acid (GABA) and taurine (TAU) contents in adult male rats and to analyze whether chronic hyperprolactinemia may affect these patterns. Rats were turned hyperprolactinemic by a pituitary graft. Plasma prolactin (PRL) levels increased after pituitary grafting at all time points examined. A disruption of the circadian rhythm was observed in pituitary-grafted rats, whereas GABA and TAU content followed daily rhythms in all areas studied in controls. In the mediobasal hypothalamus, two peaks for each amino acid were found at midnight and midday. In the anterior hypothalamus, GABA and TAU showed only one peak of concentration at midnight. In the posterior hypothalamus, the values of both GABA and TAU were higher during the light as compared to the dark phase of the photoperiod. In the median eminence GABA content peaked at 20:00h, the time when TAU exhibited the lowest values. Hyperprolactinemia abolished the 24h changes of GABA in the mediobasal hypothalamus and reduced its content as compared to controls. Hyperprolactinemia advanced the diurnal peak of TAU to 12:00h in the mediobasal hypothalamus and did not modify the 24:00h peak. In the anterior hypothalamus, hyperprolactinemia increased GABA and TAU contents during the light phase while it decreased them during the dark phase of the photoperiod. In the posterior hypothalamus hyperprolactinemia did not modify GABA or TAU patterns as compared to controls. In the median eminence hyperprolactinemia increased the 20:00h peak of GABA and shift advanced the decrease in TAU content at 20:00h and its maximum at 24:00h as compared to controls. These data show that GABA and TAU content exhibit specific daily patterns in each hypothalamic region studied. PRL differentially affects the daily pattern of these amino acids in each hypothalamic region analyzed.     

A blood plasma inhibitor is responsible for circadian changes in rat renal Na,K-ATPase activity

Rhythmic changes in activity following a circadian schedule have been described for several enzymes. The possibility of circadian changes in Na,K-ATPase activity was studied in homogenates of rat kidney cortex cells. Male Sprague-Dawley rats were kept on a schedule of 12h light (06:00-18:00 h) and 12 h darkness (18:00-06:00 h) for 2 weeks. At the end of the conditioning period, one rat was killed every 2 h, until completion of a 24 h cycle. Outermost kidney cortex slices were prepared, homogenized and assayed for Na,K-ATPase activity. The whole procedure was repeated six times. Na,K-ATPase activity shows an important oscillation (2 cycles/24 h). Peak activities were detected at 09:00 and 21:00 h, whereas the lowest activities were detected at 15:00 and 01:00-03:00 h. The highest activity was 40+/-3 nmoles Pi mg protein(-1)min(-1) (09:00 h), and the lowest was 79+/-3 nmoles Pi mg protein(-1)min(-1) (15:00 h). The amount of the Na+-stimulated phosphorylated intermediate is the same for the 09:00 h and 15:00 h homogenates. Preincubation of 09:00 h kidney cortex homogenates with blood plasma drawn from rats at either 03:00 h or 15:00 h, significantly inhibited their Na,K-ATPase activity. This inhibition was not seen when the preincubation was carried out with either 09:00 h or 21:00 h blood plasma. The striking oscillation (2 cycles/24 h) of the Na,K-ATPase activity of rat kidney cortex cells is ascribed to the presence of an endogenous inhibitor in blood plasma.     

Daily variations in the sensitivity of proestrous LH surge in the inhibitory effect of intraventricular injection of 5-HT or GABA in rats.

Intraventricular injection of 5-hydroxytryptamine (5-HT) into female rats at 11:00 h on the day of proestrus inhibited the preovulatory surge of luteinizing hormone (LH) and ovulation. A similar response was observed after the activation of the serotonergic system by stimulation of the median raphe nucleus. A diurnal rhythm of these responses was observed. In rats acclimated to a 14-h:10-h light:dark cycle the potency of 5-HT to inhibit the LH surge and ovulation was 2.06 and 2.3 times greater, respectively, when injected at 11:00 h than at 13:00 h. Also stimulation of the median raphe nucleus at 11:00 h was significantly more effective in inhibiting these parameters than stimulation at 13:00 h. Similarly, the ability of gamma-amino-butyric acid (GABA) to inhibit the preovulatory LH surge and ovulation was greater in rats injected in the morning than in the afternoon. The results of this study indicate that during proestrus the sensitivity of 5-HT and GABA to induce inhibition of preovulatory LH release and ovulation shows daily variations with maximal effect before the critical period.     

WATER DEPRIVATION AND CIRCADIAN CHANGES IN PLASMA ARGININE VASOTOCIN AND MESOTOCIN IN THE DOMESTIC HEN (GALLUS DOMESTICUS)

Possible circadian variations in plasma levels of arginine vasotocin (AVT) and mesotocin (MT) were assessed in domestic hens (Gallus domesticus) under a 12h:12h light-dark (LD) schedule. Blood samples were taken at 4h intervals, and neurohypophyseal hormone levels were determined by radioimmunoassay. Marked circadian changes in both AVT and MT were observed in hens provided free access to water. Minimal and maximal AVT levels occurred at 08:00 and 20:00, respectively. Minimal MT levels occurred at 20:00, whereas maximal MT levels occurred over a broad time period of 04:00 to 12:00. In water-deprived hens, plasma AVT levels were elevated at each time point, and the circadian variations in plasma AVT and MT levels were attenuated. These results demonstrate that rhythmicity in neurohypophyseal function in a lower vertebrate species, like that in mammals, is disrupted by osmotic stress. (Chronobiology International, 18(6), 947-956, 2001)     

模拟空间节律（L：D=0.75h：0.75h）对树qu...

It is known that contemporary space station revolves at the altitude of 400-500 km in the outer space. The present study was undertaken to investigate the effects of simulated space rhythm (L:D = 0.75 h:0.75 h) at this altitude on circadian rhythm in tree shrews (Tupaia belangeri chinensis) and the effects of endogenous sleep inducing neuropeptide Asp5-alpha-DSIP on the space-rhythm-entrained circadian rhythm. This primitive stock serves as one species of Tupaiidae and is a unique native of South China. Our previous studies have shown that this species showed striking differences in natural circadian rhythm between day and night (e.g. 3.03 degrees C of rectal temperature). Results showed that the above mentioned space rhythm (L:D = 0.75h:0.75h) could drastically disturb the inherent circadian rhythm of Tupaia belangeri chinensis. The maximal peak of motor activity dropped significantly in the morning (0600-1200) and a new enhanced peak (more than 20 times greater than that of the control) was found between 1800-2400, whereas the maximal trough of motor activity (2400-0600) remained basically unchanged. Concurrently, the total amount of 24-h motor activity was significantly decreased and the recovery after the cessation of space rhythm was slow. Experimental results also demonstrated that consecutive administration of Asp5-alpha-DSIP (30 micrograms/kg, i.p.) for 5 days (2 days before and 3 days during space rhythm) did not prevent the basic disturbance of circadian rhythm of Tupaia belangeri chinensis caused by the space rhythm (L:D = 0.75h:0.75h). Nevertheless, no decrease or even some enhancement of the total amount of 24th motor activity was observed during space rhythm or after its cessation.