Higher heart rate of laboratory mice housed individually vs in pairs

Many studies have shown that housing mice individually over a long period significantly alters their physiology, but in most cases measurement has required human interference and restraint for sampling. Using a radio-telemetry system with implantable transmitters, we recorded heart rate (HR), motor activity (ACT) and body temperature (BT) of freely moving male mice (NMRI) housed either individually or in pairs with an ovarectomized female. Data for each parameter were collected at 5 min intervals for two consecutive 24 h periods. Even after several weeks of habituation to the social conditions, HR was increased in mice housed individually compared with mice housed in pairs, although their measured ACT did not differ. Additionally, BT tended to be reduced in individually-housed mice. When the data were analysed according to different ACT levels, HR was increased in individually-housed mice during phases of low and high, but not intermediate, motor activity. Furthermore, individually-housed mice had more, but shorter, resting bouts, indicating disruption of the normal circadian sleep pattern. Enhanced HR in individually-housed mice does not necessarily indicate stress, but might be an important physiological indicator of discomfort. The fact that individual housing alters basic physiological parameters in laboratory mice highlights the need to control for housing-dependent variation, especially in experiments that are sensitive to changes in these parameters.     

Temperament moderates the influence of periadolescent social experience on behavior and adrenocortical activity in adult male rats

Adolescence is a period of significant behavioral and physiological maturation, particularly related to stress responses. Animal studies that have tested the influence of adolescent social experiences on stress-related behavioral and physiological development have led to complex results. We used a rodent model of neophobia to test the hypothesis that the influence of adolescent social experience on adult behavior and adrenocortical function is modulated by pre-adolescent temperament. Exploratory activity was assessed in 53 male Sprague–Dawley rats to classify temperament and then they were housed in one of the three conditions during postnatal days (PND) 28–46: (1) with familiar kin, (2) with novel social partners, or (3) individually with no social partners. Effects on adult adrenocortical function were evaluated from fecal samples collected while rats were individually-housed and exposed to a 1-hour novel social challenge during PND 110–114. Adolescent-housing with novel or no social partners led to reduced adult glucocorticoid production compared to adolescent-housing with familiar littermates. Additionally, highly-exploratory pre-weanling rats that were housed with novel social partners during adolescence exhibited increased exploratory behavior and a more rapid return to basal glucocorticoid production in adulthood compared to those housed with familiar or no social partners during adolescence and compared to low-exploratory rats exposed to novel social partners. In sum, relatively short-term adolescent social experiences can cause transient changes in temperament and potentially longer-term changes in recovery of glucocorticoid production in response to adult social challenges. Furthermore, early temperament may modulate the influence of adolescent experiences on adult behavioral and adrenocortical function.     

Alcohol:NAD oxidoreductase in brain of rats from a colony fed dilute ethanol for many generations.

—Alcohol:NAD oxidoreductase (EC 1.1.1.1) was studied in brain cortex, hypothalamus, cerebellum and midbrain of adult and immature rats, and in the whole encephalon of neonatal rats. The rats used in this study were (i) from a colony which has been given 12% (v/v) aqueous ethanol as the only fluid for 54 generations (‘E.F.’ rats); (ii) rats removed from this colony after the forty-eighth generation and thereafter fed water instead of the alcohol solution (‘E.F./H₂O’ rats); and (iii) normal rats. Enzyme activity in the 20,000 g supernatant of tissue homogenates was measured by the method of Raskin and Sokoloff. Activity was found to be highest in neonatal rat brain and to decrease as the age increased. Activity in the hypothalamus of adult E.F. rats was significantly higher than that found in the same region of adult E.F./H₂O rats. Immature rat cerebellum alcohol:NAD oxidoreductase activity was higher both in ‘E.F.’ and ‘E.F./H₂O’ suggesting a possible genetic change be involved in this CNS region. It may be concluded that, with the exception of neonatal rats, ethanol consumption induces an increase in rat CNS alcohol :NAD oxidoreductase activity.     

Hypothalamic levels of LHRF in male mice of differing social status and from individual housing

Swiss male mice were individually-housed or maintained in groups of 8 from weaning until 80 days of age. At this time, the grouped mice were allocated to new groups of 5–6 previously unfamiliar individuals. Identified dominants were then caged with two clearly submissive males for 1 week. At this time hypothalamic samples were taken from the dominant and one subordinate and from a number of long-term individually-housed males. A radio- immunoassay for LHRF was carried out. The levels of this hypothalamic releasing factor were significantly higher in both dominant and subordinate grouped mice than in ‘isolates’. Mice of differing social status showed comparable titres of this hormone. The results suggest that social conflict and/or disturbance augments this factor in mice irrespective of the social status achieved by individual animals.     

Threshold and receptor reserve in the action of 5-hydroxytryptamine on the salivary glands of Calliphora erythrocephala

The interrelationships of 5-HT receptors and the increased fluid secretion by isolated salivary glands of Calliphora have been studied using pharmacological techniques. Removal of the 5-OH group (tryptamine) or displacement to position 6 (6-HT) results in a decrease in potency but no change in intrinsic activity of the hormone whereas altering the ethyl amine side chain (5-OH tryptophan) results in a decrease in both potency and intrinsic activity. Removal of the 5-OH group and alteration of the side chain (gramine and tryptophan) results in a total loss of activity. Gramine and tryptophan behave as competitive antagonists of 5-HT.Prostaglandin E₁ (PGE₁) was found to be a non-competitive inhibitor of 5-OH tryptophan and theophylline whereas the response to 5-HT and cyclic AMP was only slightly diminished indicating a ‘receptor reserve’ for 5-HT.Saturating concentrations of gramine and tryptophan potentiate theophylline revealing a ‘threshold’ for the mediation of the response. It is concluded that 5-HT derivatives are capable of producing graded effects on adenyl cyclase both above and below the range of enzyme activity which evokes graded changes in fluid secretion.     

Fluoroimidazoles as antiviral agents and inhibitors of polynucleotide biosynthesis.

Some recent studies relating the effects of individual housing (isolation) and group housing to behavior, physiology and neurochemistry in laboratory rats and mice are reviewed and these accounts related to comparable information derived from experiments employing “social stresses” e.g. subjecting the animal to defeat. The data is discussed in relation to the problem of whether individual housing constitutes a “stress” (in terms of adrenocortical and adrenal medullary functioning) in these species, as it appears to do in primates. In spite of the large number of papers which ascribe the behavioral and endocrine changes obtained in isolation versus grouping comparisons to the effects of “the isolation-induced stress syndrome”, it is concluded that, in terms of adrenal function, there is little evidence that isolation $\text{per}$$\text{se}$ constitutes a stress in rats and mice, although there is some evidence that adrenocortical reactivity is increased by housing animals in this manner. It should be noted that the wild progenitor of the laboratory strains of house mouse often appears to evidence territoriality. The view is advanced that the isolated condition in male mice may result in changes characteristic of territorial dominance. This may represent a mechanism for inducing social or territorial stability in this species. It appears that experiments involving physical isolation in laboratory rodents are unlikely to provide good models for the effects of “social deprivation” in man. It is thought that more studies employing measurement of hormone titers in biological samples, obtained as a result of non-stressful procedures, will lead to a clearer understanding of the effects seen in isolation versus grouping studies. Organ weight studies often appear to be very misleading, particularly in female rodents. It is also hoped that other studies will concern themselves with effects of isolation with respect to other social cues in rodents including odors and ultrasound.     

The role of asymmetries in animal contests

This paper contains a game theoretical analysis of animal contest situations which are asymmetric in more than one aspect: two opponents may for example be imagined which differ in ‘ownership status’ as well as in ‘relative fighting ability’. The following question is analysed: which aspect may or must be used for conventional settlement in a population ‘playing’ an evolutionarily stable strategy (ESS)? The contestants are assumed to be fully informed about the asymmetric features. In particular, the assessment of relative fighting ability is supposed to be unambiguous and without cost. This assumption of perfect information allows for a decomposition of the ‘evolutionary game’ into sub-games. Therefore an easy procedure for calculating the ESS's can be presented, and simple models are analysed. It is concluded that payoff-irrelevant aspects may be used for conventional settlement of a conflict even if payoff-relevant asymmetric aspects also exist. One of the aspects may, however, be of such strong relevance that, no matter which ESS is played, animals must base their decisions on that ‘dominant’ aspect. It may also occur that two different asymmetric features are each of strong payoff relevance for either of the opponents, such that they have no escalation-suppressing effect. The particular scenario of a conflict between an ‘owner of a resource’ and an ‘intruder’ is used to derive the more general conclusions.     

Contribution of social isolation, restraint, and hindlimb unloading to changes in hemodynamic parameters and motion activity in rats

The most accepted animal model for simulation of the physiological and morphological consequences of microgravity on the cardiovascular system is one of head-down hindlimb unloading. Experimental conditions surrounding this model include not only head-down tilting of rats, but also social and restraint stresses that have their own influences on cardiovascular system function. Here, we studied levels of spontaneous locomotor activity, blood pressure, and heart rate during 14 days under the following experimental conditions: cage control, social isolation in standard rat housing, social isolation in special cages for hindlimb unloading, horizontal attachment (restraint), and head-down hindlimb unloading. General activity and hemodynamic parameters were continuously monitored in conscious rats by telemetry. Heart rate and blood pressure were both evaluated during treadmill running to reveal cardiovascular deconditioning development as a result of unloading. The main findings of our work are that: social isolation and restraint induced persistent physical inactivity, while unloading in rats resulted in initial inactivity followed by normalization and increased locomotion after one week. Moreover, 14 days of hindlimb unloading showed significant elevation of blood pressure and slight elevation of heart rate. Hemodynamic changes in isolated and restrained rats largely reproduced the trends observed during unloading. Finally, we detected no augmentation of tachycardia during moderate exercise in rats after 14 days of unloading. Thus, we concluded that both social isolation and restraint, as an integral part of the model conditions, contribute essentially to cardiovascular reactions during head-down hindlimb unloading, compared to the little changes in the hydrostatic gradient.     

A possible mechanism for the increased oxidation of choline after chronic ethanol ingestion

An attempt has been made to determine the location of the site at which the metabolism of ethanol interacts with that of choline to produce an increase in the oxidation of choline. The first enzyme in the oxidation pathway for choline, choline dehydrogenase, was assayed using a newly developed spectro-photometric assay and freshly isolated intact rat liver mitochondria. No changes were observed in either the ‘apparent’ V or the ‘apparent’ K_m values of choline dehydrogenase for choline after ethanol ingestion. However, when the choline oxidase system was assayed, a 28% decrease in ‘apparent’ K_m for choline and a 53% increase in ‘apparent’ V was observed. The effects of ATP on choline oxidase were studied further, and a 29.4% decrease was observed in mitochondrial ATP levels from freshly isolated mitochondria from the ethanoltreated rats. In vitro aging of mitochondria further decreased the level of ATP, and the rate of decrease was considerably faster during the first hour in the mitochondria from the ethanol-treated animals. The decreases in ATP from both control and experimental mitochondria were accompanied by increases in choline oxidase activity. The initial decrease in ATP was correlated with an increase in mitochondrial ATPase activity which may be related to an increase in mitochondrial Mg²⁺. Because chronic ethanol ingestion has resulted in decreased oxidation rates of succinate and β-hydroxybutyrate while at the same time increasing the oxidation rates of choline, the studies reported here suggest that the effect of chronic ethanol ingestion is primarily on a step that is unique to choline and which probably exists prior to the electron transport chain.     

The ultrastructure of the uredospore of Puccinia recondita

Dry, fresh uredospores ofPuccinia recondita that have been killed by infra-red radiation showed no striking ultrastructural differences from dry, fresh, viable uredospores. Numerous spherical and elipsoidal mitochondria with distinct and deep cristae, ribosomes, endoplasmic reticulum and convoluted plasmalemma were well shown by both. When moistened and incubated, killed uredospores lost ultrastructural organization, whereas moistened, viable, fresh uredospores imbibed moisture, became more spherical and germination commenced. A more or less centrally located nucleus was seen with the double membrane invaginated at some points. The advance of the germ tube was initially by enzymic degradation and concluded by mechanical disruption of the degraded pore plug. The mitochondria and the endoplasmic reticulum increased in number and the stored lipid bodies were gradually depleted as germination progressed. Features known as the ‘foamy cytoplasm’ and ‘folded membranes’ were seen in the germinating uredospores only. It was suggested that the ‘foamy cytoplasm’ could be functionally similar to the glyoxisome because of the close association of the former to lipid bodies. The ‘folded membranes’ may be accumulated endoplasmic reticulum being transported to the site of wall formation.     

Uptake of Amino Acids by Brain Micro vessels Isolated from Rats with Experimental Chronic Renal Failure

The neurological disorders seen in patients with chronic renal failure and liver cirrhosis are analogous. Previous in vivo studies have shown that the impaired blood-brain amino acid transport seen in rats with chronic renal failure is similar to that of rats with portocaval anastomosis. To elucidate whether a comparable underlying pathogenic mechanism plays a role in both pathological conditions, blood and brain amino acid levels together with amino acid transport by isolated brain microvessels have been studied in rats with chronic renal failure and in sham-operated rats. Brain microvessels isolated from rats with experimental chronic renal failure showed that the uptake of labeled large neutral amino acid, i.e., leucine or phenylalanine, but not of lysine or alpha-methylaminoisobutyric acid, was significantly increased with respect to sham-operated rats; conversely, the uptake of glutamic acid in rats with chronic renal failure was significantly lower compared with values in controls. Kinetic analysis indicated that this was mainly due to increased exchange transport activity (Vmax) of the L-system, rather than to changes in the affinity (Km) of the carrier system for the relative substrate. These data, together with the significant rise of brain glutamine levels and an increased brain-to-plasma ratio of the sum of large neutral amino acids, are analogous to what was previously observed in rats with portocaval anastomosis.(ABSTRACT TRUNCATED AT 250 WORDS)     

Activity of the hypothalamic–pituitary–adrenal axis of prenatally stressed male rats in experimental model of depression

Changes in activity of the hypothalamic–pituitary–adrenal (HPA) axis were examined in adult, prenatally stressed male rats in the experimental depression model of ‘learned helplessness’. It was shown that in males descending from intact mothers a depressive-like state was accompanied by an increase in activity of the entire HPA axis. Namely, expression of corticotropin-releasing hormone (CRH) in the hypothalamic paraventricular nucleus (PVN) increased coupled to a rise in plasma levels of ACTH and corticosterone as well as in adrenal weight. At the same time, in males born to mothers who suffered stress during the last week of pregnancy a decrease was detected in activity both of the central (hypothalamus) and peripheral (adrenal cortex) parts of this regulatory hormonal axis, analogous to that we revealed previously in the ‘stress–restress’ experimental model. It is concluded that prenatal stress modifies the sensitivity of animals to inescapable intense stress impacts, as manifested in the specific pattern of HPA axis activity after stressing.     

Rapid hyperthyroidism-induced adaptation of salmonid fish in response to environmental pollution

The streams draining volcanic landscapes are often characterized by a complex series of factors that negatively affect hydrobionts and lead to declines in their populations. However, in a number of cases, a range of rapid adaptive changes ensure the resilience of hydrobiont populations. Here, we present both field and experimental data shedding light on the physiological basis of adaptation to heavy metal contamination in populations of Dolly Varden charr (Salvelinus malma) differing in duration of isolation in volcanic streams. The study reveals that isolated populations have a physiological phenotype that distinguishes them from populations inhabiting clean waters. They are characterized by a hyperthyroid status accompanied by an increased metabolic rate, elevated activity of antioxidant enzymes, decreased ionic conductivity of tissues and reduced stored energy reserves. Our experimental data reveal that hyperthyroidism is an adaptive characteristic enhancing the resistance to heavy metal contamination and shaping the evolution of these populations. The similarity of physiological, developmental and morphological changes in isolated populations suggests a common source and mechanisms underpinning this case of ‘evolutionary rescue’. Thus, populations of S. malma trapped in volcanic streams represent a genuine case of rapid endocrine-driven adaptation to changing environmental stimuli.     

Increased illumination level during gestation and offspring emotionality in the rat

Open-field defecation, ambulation and HR were recorded from a group of rats whose mothers had been exposed to high intensity illumination during pregnancy. Compared with controls, experimental animals showed reduced defecation, increased HR, and no significant differences in ambulation. No significant sex differences in open-field measures were found. It was concluded that illumination level is a more satisfactory independent variable in prenatal maternal stress studies, than the traditional ‘conditioned anxiety’ technique. The findings were compared with those of other studies in the area, and discussed in relation to theories of autonomic functioning in emotional behaviour.     

Isolation and characterisation of DNA cytosine 5-methyltransferase from human placenta

DNA cytosine 5-methyltransferase has been extensively purified (about 2600-fold) from the soft tissue of human placenta by chromatography on DEAE-cellulose and hydroxyapatite, and by an affinity step on agarose-immobilized S-adenosylhomocysteine. The isolated enzyme has a molecular weight of 135000 and methylates DNA from various sources in native and heat-denatured forms. The synthetic copolymer poly(dG-dC)·poly(dG-dC) is methylated in B- and Z-conformation to about the same extent. DNA containing hemimethylated sites was isolated from P815 cells grown in the presence of 5-azacytidine. This P815 DNA was used to measure the ‘maintenance’ DNA methylase activity, whereas 5-methylcytosine-free procaryotic DNA served as a substrate for the ‘de novo’ DNA methylase activity in our enzyme preparation. The crude extract as well as the highly purified DNA methylase are capable of transferring methyl groups to these two types of substrate. The fact that both types of activity co-chromatograph during the isolation procedure suggests that one enzyme molecule may exercise both the ‘maintenance’ and ‘de novo’ activity.     

Further characterization of the effects of 3-methylcholanthrene administration upon hepatic ribonucleic acid polymerase activities

The administration of 3-methylcholanthrene (MC) to rats results in a marked increase in the specific activities of hepatic RNA polymerases I and II. In the present study, we were able to show that this increase was not caused by a shift in the ratio of ‘free’ to ‘template-engaged’ RNA polymerase. By means of binding studies with [³H]amatoxin, we were unable to demonstrate any increase in the number of RNA polymerase II molecules in liver after MC administration to the rats. RNA polymerase I was purified in excess of 3000-fold from hepatic nuclei isolated both from control and MC-treated rats. The stimulation in activity was demonstrated at each step in the purification scheme until glycerol sedimentation analysis. Results from cation-exchange chromatography on phosphocellulose indicated that the polycyclic hydrocarbon increased the enzyme activity of RNA polymerase I_b somewhat specifically. Subsequent to glycerol gradient centrifugation, this stimulatory advantage was no longer evident. Reconstitution experiments revealed the presence of a stimulatory component, which was demonstrated in low molecular weight fractions from both control and experimental preparations.     

Effects of predictability on the welfare of captive animals

Variations in the predictability of a stressor have pronounced effects on the behavioural and physiological effects of stress in rats. It is reasonable to expect that variations in the predictability of husbandry routines thought to be aversive to animals might have similar effects on stress indices. Similarly, variations in the predictability of positive events, of which feeding is an obvious example, may affect welfare. This review examines the behavioural and physiological effects of the predictability of aversive and appetitive stimuli, and the application of experimental findings to animal husbandry in practice. It is argued here that two distinct but overlapping types of predictability exist. ‘Temporal’ predictability describes whether an event occurs at fixed or variable intervals, whereas ‘signalled’ predictability relates to the reliability of a signal preceding the event. This review examines the effects of each of these types of predictability in relation to positively and negatively perceived events, and examines the link between predictability and control. Recommendations are made for relatively simple and inexpensive modifications to husbandry routines that may be easy to incorporate into the schedules of busy staff yet could have a profound impact on the welfare of animals in their care.     

Western diet given to healthy rats mimics the human phenotype of diabetic cardiomyopathy

Diabetes mellitus (DM) is a major problem worldwide. Within this patient group, cardiovascular diseases are the biggest cause of morbidity and mortality. Diabetic cardiomyopathy (DCM) is defined as diabetes-associated structural and functional changes in the myocardium, not directly attributable to other confounding factors such as coronary artery disease or hypertension. Pathophysiology of DCM remains unclear due to a lack of adequate animal models reflecting the current pandemic of diabetes, associated with a high increased sugar intake and the ‘Western’ lifestyle. The aim of this study was to develop an animal model mimicking this ‘Western’ lifestyle causing a human-like phenotype of DCM. Twenty-four Sprague–Dawley rats were randomly assigned into a normal or a ‘Western’ diet group for 18 weeks. Glucose and insulin levels were measured with an OGTT. Heart function was assessed by echocardiography and hemodynamic measurements in vivo. Cardiac fibrosis and inflammation were investigated in vitro. ‘Western’ diet given to healthy rats for 18 weeks induced hyperglycemia together with increased AGEs levels, insulin levels and hypertriglyceridemia. Heart function was altered with increased end-diastolic pressure, left ventricle hypertrophy. Changes in vivo were associated with increased collagen deposition and increased PAI-1 levels in the heart. High-sugar diet or ‘Western’ diet causes T2DM and the hallmarks of DCM in rats, reflecting the phenotype of the disease seen in patients. Using this new model of T2DM with DCM might open new insight in understanding the pathophysiology of DCM and on a long term, test targeted therapies for T2DM with DCM patients.     

Changes in short‐chain acyl‐coA dehydrogenase during rat cardiac development and stress

This study was designed to investigate the expression of short‐chain acyl‐CoA dehydrogenase (SCAD), a key enzyme of fatty acid β‐oxidation, during rat heart development and the difference of SCAD between pathological and physiological cardiac hypertrophy. The expression of SCAD was lowest in the foetal and neonatal heart, which had time‐dependent increase during normal heart development. In contrast, a significant decrease in SCAD expression was observed in different ages of spontaneously hypertensive rats (SHR). On the other hand, swim‐trained rats developed physiological cardiac hypertrophy, whereas SHR developed pathological cardiac hypertrophy. The two kinds of cardiac hypertrophy exhibited divergent SCAD changes in myocardial fatty acids utilization. In addition, the expression of SCAD was significantly decreased in pathological cardiomyocyte hypertrophy, however, increased in physiological cardiomyocyte hypertrophy. SCAD siRNA treatment triggered the pathological cardiomyocyte hypertrophy, which showed that the down‐regulation of SCAD expression may play an important role in pathological cardiac hypertrophy. The changes in peroxisome proliferator‐activated receptor α (PPARα) was accordant with that of SCAD. Moreover, the specific PPARα ligand fenofibrate treatment increased the expression of SCAD and inhibited pathological cardiac hypertrophy. Therefore, we speculate that the down‐regulated expression of SCAD in pathological cardiac hypertrophy may be responsible for ‘the recapitulation of foetal energy metabolism’. The deactivation of PPARα may result in the decrease in SCAD expression in pathological cardiac hypertrophy. Changes in SCAD are different in pathological and physiological cardiac hypertrophy, which may be used as the molecular markers of pathological and physiological cardiac hypertrophy.