Photic and non-photic effects on the daily activity pattern of Mongolian gerbils.

The paper analyses the daily activity pattern of Mongolian gerbils with and without access to a running wheel. To evaluate the synchronizing and the masking effects of light, experiments were performed under different photoperiods (L:D=14:10 h and 10:14 h), and light and dark pulses were applied at different phases of the day-night cycle. In order to get a more direct estimate of the central pacemaker of the circadian system, the body temperature rhythm was investigated via implanted transmitters. Without access to a running wheel, the daily activity pattern was bimodal. One peak occurred in the first half of the light time, the other one around the light-dark transition. Also, the gerbils were more active during the light phase as compared to the dark phase. After unlocking the running wheel, the gerbils were active mainly during the dark time. The activity peak in the first half of the light phase remained, the second one shifted by a phase delay into the dark time. These results were found under both LD-regimens. Light during the night nearly completely suppressed running wheel activity, switching off the light during the day time induced wheel running. Whereas wheel running was clearly affected by light and dark pulses, the general activity was not. The body temperature rhythm also shows two peaks, with the second one being bigger and coinciding with the endogenous component of the circadian body temperature rhythm. It was found around light-off. After unlocking the running wheel, the maximum of the body temperature rhythm shifted to the night. This was not primarily a consequence of the changed activity pattern as shown by means of purification analysis. Removing the direct effects of motor activity led to a body temperature curve that could be described by a cosine function, and the delay shift was found also for the purified data, a better estimate of the endogenous circadian component. The wheel-associated increase in nocturnality is not only due to masking effects of wheel-running activity on the body temperature and activity rhythms. It also involves clock-related processes. Changes in the phase preference may serve as an adaptation mechanism to the changes in the animal's natural environment.

Influence of oxygen concentration on redox cycling of alloxan and dialuric acid.

Alloxan, a chemical diabetogen, decays in the absence of reductants into alloxanic acid. In the presence of glutathione, it is reduced via the alloxan radical into dialuric acid, which autoxidizes back to alloxan. During this redox cycling process, reactive oxygen species are formed that destroy beta-cells in islets of Langerhans. Previous experiments were conducted with oxygen concentrations about ten times as high as within cells. The aim of our in vitro study was to evaluate the impact of different oxygen concentrations (0, 25, 250 micromol/l) at a given initial ratio of glutathione and alloxan on this redox cycling. Reduction of alloxan, oxidation of glutathione, and the formation of glutathiol (GSSG) were continuously recorded by HPLC for 90 minutes at 25 degrees C in air, calibration gas, or argon. In the absence of reductants, alloxan irreversibly decomposed into alloxanic acid regardless of oxygen presence. When the reaction system contained glutathione, decomposition was significantly retarded and therefore influenced by oxygen. In argon, decay could not be observed due to its reduction and the absence of oxygen. Increasing oxygen concentration enabled a redox cycling and therefore an ongoing decay. The highest decomposition along with the highest consumption of glutathione occurred at 250 micromol/l oxygen. The lower the oxygen, the more dialuric acid could be detected. After calculation, about 33 redox cycles per hour generates an amount of reactive oxygen species sufficient to damage pancreatic beta cells and induce insulin deficiency.

Indicators for post-surgery recovery in Mongolian gerbils (Meriones unguiculatus).

Telemetric investigations of various parameters are widely used to estimate an animal's state. However, the implantation of the transmitters includes anaesthesia and surgery and has short and longer lasting impacts on the studied object. The aim of the present paper was to evaluate these effects in Mongolian gerbils, namely the hypothermia caused by the anaesthetic and the enduring disturbance of daily rhythmicity until complete recovery. The surgery associated with the implantation of the transmitters differed both in severity and type of anaesthesia. Whereas normal values of body temperature were restored within hours, restoration of daily rhythm required several days, depending on the severity of the surgical procedure. Also, the sensitivity of the body temperature to activity changes was different until the rhythms were re-established. A method based on the rhythm magnitude and shape was proposed to estimate the time until complete recovery of the animals.

The development of new purification methods to assess the circadian rhythm of body temperature in Mongolian gerbils.

Six Mongolian gerbils were studied for 8-10d while housed in separate cages in a 12:12h light-dark (L-D) cycle (lights on at 07:00h). Recordings of body temperature, heart rate, and spontaneous activity were made throughout. The temperature and heart rate rhythms were "purified" to take into account the effects of activity, and then the rhythm of temperature was further purified to take into account other masking influences ("non-activity masking effects" or NAME,). The methods employed in the purification processes involved linear regression analysis or analysis of covariance, the latter using functions of activity and NAME as covariates. From these methods, it was possible to obtain not only an estimate of the endogenous component of the temperature rhythm but also a measure of circadian changes in the sensitivity of temperature to masking effects. Even though all purification methods removed many of the effects of spontaneous activity from the temperature record, there remained temperature fluctuations at the L-D and D-L transitions that appeared to be independent of activity. The NAME was of only very marginal value in the purification process. Comparison of the purification methods indicated that the linear methods were inferior (both from a biological viewpoint and when the results were compared mathematically) to those that allowed the rate of rise of temperature due to increasing amounts of activity to become progressively less. The sensitivity of temperature and heart rate to the masking effects of activity showed a circadian rhythm, with sensitivities in the resting phase being greater than those in the active phase. These findings are compatible with the view that thermoregulatory reflexes are induced by spontaneous activity of sufficient amount, and that there is a circadian rhythm in the body temperature at which these reflexes are initiated and in their effectiveness.

Comparative studies of body mass, body measurements and organ weights of wild-derived and laboratory golden hamsters (Mesocricetus auratus).

All laboratory golden hamsters originate from a sibling pairing back in 1930. To investigate possible differences between domesticated and wild conspecifics, descendants of both strains were maintained under standardized laboratory conditions individually and in unisexual groups. Body mass and food consumption were monitored from birth to 22 weeks of age. The animals were subsequently sacrificed, and body measurements and body composition were analysed. In addition, the absolute and relative masses of different organs were measured. Laboratory hamsters gained more body mass through higher food consumption. However, they did not get fatter, since relative fat values were the same for both strains. Body measurements revealed only minor differences (in body and ear lengths). As deducible from the body mass, the organs (spleen, kidneys, adrenal glands, testes, epididymis and ovaries) were seen to be heavier in laboratory hamsters. Furthermore, with the exception of the kidneys, the same went for the relative values. There were distinct sexual specific differences in both strains only for body fat ( male symbol male symbol upward arrow ) and adrenal glands ( male symbol male symbol upward arrow ). In females, group housing induced an elevated level of aggression. In general, these housing conditions led to social stress symptoms, such as heavier adrenal glands. Additionally, spleen, kidneys, ovaries, body length and mass, body water and body fat were increased in group-housed hamsters. In conclusion, no major differences between laboratory and wild-derived hamsters were observed.

Formation of compound 305 requires the simultaneous generation of both alloxan and GSH radicals.

This in vitro study investigates the conditions under which "compound 305" is formed. Using HPLC, ESR as well as UV spectroscopy, "compound 305" was largely separated and characterized. It has an absorption peak at 314 nm, which changes after reoxygenation to shorter wavelengths within hours. The retention time of "compound 305" amounts to 10.93 +/- 0.042 min. The formation of "compound 305" does not depend on alloxan (ALX) or reduced glutathione (GSH), but most likely on the steady-state concentration of the paramagnetic derivatives of both reactants (ALX* and GS*). The alloxan radical (ALX*) is formed by either a one-electron transfer from e. g. GSH to alloxan or oxidation of dialuric acid. The concentration of the ALX* was determined to be 12 +/- 3.6 micromol/l using the stable ultramarine radical as an ESR standard. ALX* is stable only under anaerobic conditions. It disappears within 2 min in air. Since formation of "compound 305" needs both ALX* as well as GS*, which are also necessary for the generation of reactive oxygen species (ROS), it is assumed that formation of "compound 305" diminishes the toxicity of alloxan.

Simultaneous quantitative determination of alloxan, GSH and GSSG by HPlc. Estimation of the frequency of redox cycling between alloxan and dialuric acid.

This in vitro study compares the frequency of redox cycling between alloxan and dialuric acid at different initial ratios of glutathione and alloxan. Alloxan oxidizes GSH to GSSG. The rate of GSH oxidation at a given initial GSH concentration of 2.0 mmol/L depends on the initial concentration of alloxan added. The higher the concentration of alloxan in relation to the initial concentration of GSH, the faster GSH oxidation proceeds, as well as oxygen consumption, and therefore, formation of reactive oxygen species. The highest rates of GSH oxidation, i.e. GSSG formation, were found at concentration ratios of between 2.0 mmol/L GSH and 0.2 and 0.04 mmol/L alloxan, respectively. Because 0.04 mmol/L alloxan oxidizes 2.0 mmol/L GSH completely, a frequency of at least 25 cycles between alloxan and dialuric acid within 3 hours can be assumed. During each redox cycle, two molecules of GSH are oxidized to one molecule of GSSG, and during each cycle one molecule of oxygen is reduced simultaneously to one molecule of hydrogen peroxide. In total, therefore, one molecule of alloxan oxidizes at least 50 molecules of GSH and forms about 25 molecules of hydrogen peroxide.

Circadian activity rhythms and sensitivity to noise in the Mongolian gerbil (Meriones unguiculatus).

Since consistent data on endogenous circadian rhythms of Mongolian gerbils are not available, the main aim of our study was to identify suitable conditions to receive stable and reproducible free-running rhythms of activity under different light intensities. Another objective was to determine the role of social cues as an exogenous zeitgeber in the absence of a light-dark (LD) cycle. We performed two long-term sets of experiments with adult male gerbils kept in climatic chambers under various photoperiods of at least 30 days each. In all cases, the time of lights on in the chambers differed from the daily starting hour of work in the animal house. Always, two animals per chamber were kept separately in cages with a running wheel while their activity was monitored continuously. During the first set, only three of eight animals developed intra- and interindividual variable free-running rhythms. The activity patterns seemed to be influenced by human activities outside, indicating high sensitivity to external factors. Subsequently, we damped the chambers and the room and restricted access to the room. In the following noise-reduced set, all gerbils developed comparable free-running rhythms of activity. We determined the mean of the free-running period tau, the activity-rest relationship alpha/theta and the amount of running wheel activity per day: tau = 23.7h +/- 0.08h under low light (5 lux) and 25.5h +/- 0.19h under high light intensities (450 lux); alpha/theta = 0.53 +/- 0.08 under 5 lux and 0.34 +/- 0.04 under 450 lux. The amount of daily activity was 12 times as high under 5 lux as under 450 lux. There was no indication that the two animals in one chamber socially synchronized each other. In conclusion, the pronounced rhythm changes in accordance with Aschoff's theory support the view that gerbils are mainly nocturnal animals.

Time of day and stress response to different stressors in experimental animals. Part I: Golden hamster (Mesocricetus auratus Waterhouse, 1839).

The present paper describes the effects of animal house routine stressors on adult golden hamsters during activity time (2 hrs after lights off) and rest time (2 hrs after lights on). In addition, for determination of norm values, the circadian rhythms of the stress indicators heart rate, core body temperature and general activity of unstressed animals were telemetrically registered via implanted transmitters. The three circadian patterns of the nocturnal golden hamster under L:D = 12:12 were unimodal with a main peak after lights off. The physiological norm values (mean over 24 hours +/-SD) were: heart rate 324 +/- 18 bpm, core body temperature 37.5 +/- 0.5 degrees C and activity 114 +/- 123 units/5 min. The mean body temperature of females was significantly higher (0.4 degree C) and its mean activity level was significantly (40%) lower than that of males. The stress responses were dependent on the time of day and on the kind of stressor. The stress responses were significantly stronger during the rest time of the animals (i.e. light period), and it resulted in the subsequent ranking of stressors: handling < vaginal smear < intruder/resident confrontation < cage changing < grouping. There were no sex-dependent stress response differences. The results of this study were compared with identical investigations on the social Mongolian gerbil (J. Exp. Anim. Sci. 1996/97; 38: No. 3).

Measurement of physiological parameters and activity in a Mongolian gerbil during gravidity and lactation with an implanted transmitter.

This communication reports the first use of a transmitter implanted into the abdominal cavity of a small pregnant rodent to simultaneously measure the heart rate, core body temperature and general locomotory activity with no apparent side effects to the animals. Using this method the heart rate of one female Mongolian gerbil (Meriones unguiculatus) decreased from a mean of 303 +/- 6 (SE) (beats per minute) during gravidity to a level of 248 +/- 3 (SE) (beats per minute) during lactation. At the same time, the core body temperature increased from 37.7 +/- 0.05 (SE) degrees C to 38.0 +/- 0.03 (SE) degrees C. During pregnancy the general locomotory activity of this female followed the normal nocturnal pattern for this species (i.e., greater activity during the dark period) but there was an overall reduction in activity to 85% of the nonpregnant state. The level of activity was further reduced during lactation to about 43% of the nonpregnant state. This was largely the result of less activity during the dark period so that activity in the dark period and light period became more or less equivalent.