Antioxidant status in the liver of hypertensive and metallothionein-deficient mice

Because oxidative stress is involved in arterial hypertension, impairment of hepatic antioxidant defences could develop in the course of this disease. Metallothionein (MT), an antioxidant protein, is present in high rates in the liver. The aim of this study was to investigate the effect of a mineralocorticoid-salt treatment on blood pressure, hepatic antioxidant enzyme activities, and cardiac MT levels in transgenic MT null mice compared with control mice to further clarify the role of MT during the experimental development of arterial hypertension. Control and transgenic MT -/- mice were submitted to an 8-week mineralocorticoid-salt treatment. Hepatic glutathione peroxidase, glutathione reductase, superoxide dismutase, and catalase activities and cardiac MT and mineral levels were measured. Mineralocorticoid-salt treatment induced an increase in blood pressure in both transgenic MT -/- and control mice that was associated with an impairment of liver antioxidant status. MT deficiency was associated with modifications of hepatic antioxidant enzyme activities and with a decrease in cardiac iron levels. Adaptive processes of antioxidant systems may explain the absence of an effect of metallothionein deficiency on the development of mineralocorticoid-salt hypertension. The interactions that occur between the in vivo antioxidant systems probably produce a complex regulation of the oxidative balance and consequently prevent antioxidant deficiency.     

Hepatic microvascular responses to ischemia-reperfusion in low-density lipoprotein receptor knockout mice

The overall objective of this study was to determine whether genetically induced hypercholesterolemia alters the inflammatory and microvascular responses of mouse liver to ischemia-reperfusion (I/R). The accumulation of rhodamine 6G-labeled leukocytes and the number of nonperfused sinusoids (NPS) were monitored (by intravital microscopy) in the liver of wild-type (WT) and low-density lipoprotein receptor-deficient (LDLr(-/-)) mice for 1 h after a 30-min period of normothermic ischemia. Plasma alanine transaminase (ALT) levels were used to monitor hepatocellular injury. Microvascular leukostasis as well as increases in NPS and plasma ALT were observed at 60 min after hepatic I/R in both WT and in LDLr(-/-) mice; however, these responses were greatly exaggerated in LDLr(-/-) mice. Pretreatment of LDLr(-/-) mice with gadolinium chloride, which reduces Kupffer cell function, attenuated the hepatic leukostasis, NPS, and hepatocellular injury elicited by I/R. Similar protection against I/R was observed in LDLr(-/-) mice pretreated with antibodies directed against tumor necrosis factor-alpha, intercellular adhesion molecule-1 (ICAM-1), or P-selectin. These findings indicate that chronic hypercholesterolemia predisposes the hepatic microvasculature to the deleterious effects of I/R. Kupffer cell activation and the leukocyte adhesion receptors ICAM-1 and P-selectin appear to contribute to the exaggerated inflammatory responses observed in the postischemic liver of LDLr(-/-) mice.     

Anxiety stress and nociceptive responses in mice

Nociception in laboratory animals appears to be influenced by physical or emotional stressors. Nevertheless, the reported data are not univocal. Discrepancies seem to be caused by some kind of stress model and/or by the timing of stressor application. The aim of the present work is to study the influence of chronic application of a well-controlled and defined anxiety stress paradigm (rotational stress) on the behavioral formalin pain responses in mice maintained in a low-stress environment. The results indicate that emotional chronic stress increases specific pain responses in the late inflammatory phase and, correspondingly, decreases self-grooming. Locomotor activity appears influenced by pain presence only. The hormonal and neural mechanisms that could be involved in the observed nonspecific and specific nociceptive responses to stress are discussed.     

Hepatic effects of phenobarbitone in female mice fed sublethal levels of dietary cyanide

Female albino mice were fed sublethal doses of KCN (approx. 10 micrograms/mouse/day) for 7 days, injected intraperitoneally with phenobarbitone (50 mg/kg body wt/day) in the subsequent 3 days, and sacrificed 24 hr after the last injection. Phenobarbitone sleeping time was increasingly shortened (16-27%) daily in cyanide-fed mice in comparison with cyanide-free controls. Both compounds administered singly or simultaneously increased the liver weight/body weight ratios by not more than 10%. Aniline hydroxylase, glucose-6-phosphatase, NADPH- and NADH-cytochrome c reductase activities were similarly increased. Aniline hydroxylase activity was most markedly increased (by a factor of 4). The toxicological implications of these results are discussed.     

Hepatic HMG-CoA reductase expression and resistance to dietary cholesterol

The premise that the intrinsic level of expression of hepatic 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase determines the relative sensitivity to the serum cholesterol raising action of dietary cholesterol was examined in 9 strains of rat. For further comparison purposes, hamsters were also examined. The basal expression of hepatic HMG-CoA reductase, extent of feedback regulation by cholesterol, and changes in serum cholesterol levels and the hepatic low-density lipoprotein (LDL) receptor in response to cholesterol challenge were determined in these animals. The Sprague-Dawley, Wistar-Furth, Spontaneously Hypertensive, Lewis, and Wistar-Kyoto rats were all very resistant to dietary cholesterol and exhibited hepatic HMG-CoA reductase activities above 150 pmol / min(-1) / mg(-1). The Buffalo, Brown Norway, and Copenhagen 2331 rats had hepatic HMG-CoA reductase activities below 90 pmol / min(-1) / mg(-1) and had increases in serum cholesterol levels ranging from 12 to 33 mg/dl when given a 4-day, 1% cholesterol challenge. The extent of feedback regulation was reduced to only 3-fold in the Fisher 344 and Brown Norway rats that exhibited significant increases in serum cholesterol levels when given a cholesterol challenge. The Golden Syrian hamsters exhibited the largest increase (197 mg/dl) in serum cholesterol levels in response to dietary cholesterol and the lowest basal expression of hepatic HMG-CoA reductase (3.3 pmol / min(-1) / mg(-1)). Hepatic LDL receptor levels were not significantly decreased by dietary cholesterol in any of the animals. The data from these inbred rats and the hamsters strongly support the conclusion that the animals expressing the highest levels of hepatic HMG-CoA reductase are the most resistant to the serum cholesterol raising action of dietary cholesterol.     

Estrogen, insulin, and dietary signals cooperatively regulate longevity signals to enhance resistance to oxidative stress in mice

To investigate the biological significance of a longevity mutation found in daf-2 of Caenorhabditis elegans, we generated a homologous murine model by replacing Pro-1195 of insulin receptors with Leu using a targeted knock-in strategy. Homozygous mice died in the neonatal stage from diabetic ketoacidosis, whereas heterozygous mice showed the suppressed kinase activity of the insulin receptor but grew normally without spontaneously developing diabetes during adulthood. We examined heterozygous insulin receptor mutant mice for longevity phenotypes. Under 80% oxygen, mutant female mice survived 33.3% longer than wild-type female mice, whereas mutant male mice survived 18.2% longer than wild-type male mice. These results suggested that mutant mice acquired more resistance to oxidative stress, but the benefit of the longevity mutation was more pronounced in females than males. Manganese superoxide dismutase activity in mutant mice was significantly upregulated, suggesting that the suppressed insulin signaling leads to an enhanced antioxidant defense. To analyze the molecular basis of the gender difference, we administered estrogen to mutant mice. It was found that the survival of mice under 80% oxygen was extended when they were administered estradiol. In contrast, mutant and wild-type female mice showed shortened survivals when their ovaries were removed. The influence of estrogen is remarkable in mutant mice compared with wild-type mice, suggesting that estrogen modulates insulin signaling in mutant mice. Furthermore, we showed additional extension of survival under oxidative conditions when their diet was restricted. Collectively, we show that three distinct signals; insulin, estrogen, and dietary signals work in independent and cooperative ways to enhance the resistance to oxidative stress in mice.     

Autonomic responses to stress

Forty-eight male subjects with no previous meditative experience engaged in either progressive relaxation (PR), a meditative treatment designed to induce the relaxation response (RR), or a no-treatment control experience (C) during four sessions on consecutive days. Negative expectations regarding the effectiveness of each technique for reducing physiological responses to stress were induced for half of the subjects in each treatment condition, and positive expectations were induced for the other half. Subjects viewed a stressful film following practice of their technique during the first and fourth sessions. Heart rate and electrodermal responding were recorded continuously during practice of the techniques and during the stressful film throughout the first and fourth sessions. Results indicated lowered heart rate levels prior to the film for subjects in the PR-positive expectancy condition and during the film for subjects in the RR-positive expectancy condition. It is suggested that subjects' expectancies concerning meditation may affect cardiovascular responding during stress, although meditative treatments in general do not appear to reduce stress responding as effectively as previously suggested.This research is based upon a master's thesis submitted by the first author to the graduate school at Northern Illinois University. The research was supported in part by a grant to the second author from the graduate school at Northern Illinois University. Portions of the article were presented at the convention of the Midwestern Psychological Association in Chicago, 1978.     

Endorphins do not affect behavioral stress responses in mice.

Effects of endorphins on behavioral stress responses were investigated in mice. For this purpose, we used environment-induced conditioned suppression of motility and forced swimming-induced immobility. The cerebral ventricular administration of alpha-endorphin (2.5-10 nmol), beta-endorphin (0.38-1.5 nmol), or gamma-endorphin (2.5-10 nmol) failed to affect either the environment-induced conditioned suppression of motility or the forced swimming-induced immobility. We have indicated previously that enkephalins attenuate both stress responses and, in contrast, dynorphin potentiates them. These findings indicate that the endorphinergic systems are not responsible for behavioral stress responses and that the role played by endorphins in the present stressful situations may be different from that of enkephalin and dynorphin.     

Side effects of control treatment can conceal experimental data when studying stress responses to injection and psychological stress in mice

Routine laboratory procedures, such as handling or transporting animals or carrying out injections on animals, are stressful for animals but are necessary in many pre-clinical studies. Here, the authors show that multiple injections of the non-toxic vehicle cyclodextrin moderately increased plasma corticosterone concentrations in female BALB/c mice. Additionally, male BALB/c mice that had received a single intraperitoneal injection of harmless saline had an increased glucocorticoid response to a second saline injection. The authors found that female mice that had been exposed to an acute psychological stress session had a decreased glucocorticoid response to a second homotypic stressor. In contrast, multiple psychological stress sessions led to increased glucocorticoid release in female mice. Acute injection(s) of saline in male mice and of cyclodextrin in female mice led to transient lymphocytopenia. Further analysis showed that repeated stress-induced lymphocytopenia is glucocorticoid-dependent. The authors conclude that laboratory stress can affect physiological parameters in mice, potentially altering study results.     

Ethylene and plant responses to stress

When plants are subject to a variety of stresses they often exhibit symptoms of exposure to ethylene. Although this relationship usually results from induction of ACC synthase thus raising the concentration of the precursor of ethylene, it is now apparent that there are numerous other ways that stresses produce ethylene-like symptoms. This complex relationship between stress and ethylene-like symptoms is here termed the stress ethylene syndrome. ACC synthase exists as a multi-gene family whose individual members are differentially regulated, many by various stresses. In addition, ACC oxidase. AdoMet synthetase, enzymes in the Yang methionine cycle, and enzymes that conjugate ACC are regulated by stress. In more unusual cases, ethylene production is not increased by stress or may be reduced. There is evidence for stress effects on perception of ethylene and the potential exists that some steps of the ethylene signal transduction pathway may be influenced by stress. Because of the variability possible in the stress ethylene syndrome, it continues to be studied for a number of stresses and species. In particular, attention is being given to wounding, mechanical stress, drought, heat and water deficit stress, chilling, air pollution, chemical and salt stress, and low O₂stress. It is becoming more apparent that a number of stress responses involve interactions with other hormones.     

Paternal experience and stress responses in California mice (Peromyscus californicus)

Paternal behavior greatly affects the survival, social development, and cognitive development of infants. Nevertheless, little research has been done to assess how paternal experience modifies the behavioral characteristics of fathers, including fear and stress responses to a novel environment. We investigated long-term behavioral and physiologic effects of parental experience in mice (Peromyscus californicus) and how this response activates the hypothalamic-pituitary-adrenal axis (as measured by corticosterone and dehydroepiandrosterone [DHEA] levels) and interacts with anxiety-related behaviors. Three groups of adult males were tested--fathers exposed to pups, virgins exposed to pups, and virgins never exposed to pups--in 2 environments designed to elicit anxiety response: an open field with a novel object placed in the center and a closed cage containing a sample of a component of fox feces. Behavioral responses were measured by using traditional methods (duration and frequency) and behavioral-chain sequences. Results indicated that paternal experience significantly modifies a male mouse's behavioral and physiologic responses to stress-provoking stimuli. Compared with inexperienced male mice, experienced male mice had a significant decrease in the occurrence of incomplete behavioral chains during the exposure to the novel object, an index of reduced stress. Further, even moderate pup exposure induced behavioral modifications in virgin male mice. These behavioral responses were correlated with changes in corticosterone and DHEA levels. Together, these data provide evidence that interactions between male mice and offspring may have mutually beneficial long-term behavioral and physiologic effects.     

Rapid responses to stress in Eurytemora affinis

E. affinis can adjust to temperature stress in a matter of hours. Adaptation is greater in a varying temperature than in a constant temperature, consistent with the estuarine habitat of this calanoid. The species has the capacity to adjust both in the short-term as individuals and also genetically over a number of generations. The adjustments have been examined at several levels of organization. In whole copepods the time an individual becomes comatose when exposed to a 32 °C temperature and increasing by 1/2 °C at 5 min. intervals, has been used as a repeatable assay and gives a good prediction of survival at 30 °C, the ecological limit of the species in Chesapeake bay, USA. At the molecular and cellular levels, two adaptive mechanisms which have been observed in temperature stressed copepods are the synthesis of novel proteins and phase changes in plasma membrane lipids. Both of these mechanisms have potential for further understanding the adaptation of Eurytemora to variable temperatures. They may also have application as indicators of sublethal stress.