Cardioprotective Effects of Mebudipine in a Rat Model of Doxorubicin-Induced Heart Failure.

Background: Mebudipine, a dihydropyridine calcium-channel blocker (CCB), shows greater time- and voltage-dependent inhibitory effects than nifedipine. Its significant negative chronotropic effects without having considerable negative inotropic properties may make it a suitable candidate for the pharmacotherapy of heart failure (HF). This study aimed to investigate the possible beneficial action of mebudipine in a rat model of HF. Methods: The present study carried out in the Department of Pharmacology at the Iran University of Medical Sciences during the years of 2009-2011. An experimental model of HF was induced in male Wistar rats using doxorubicin (DOX). The rats were divided into five groups with seven animals in each group: normal control group, DOX-induced HF control groups, and treatment groups. The animals were administered DOX for 15 days. A consistent deterioration occurred after a four-week rest period. The animals were then treated with intraperitoneal mebudipine (0.5 mg/kg) and intraperitoneal amlodipine (0.35 mg/kg), as well as an equal volume of distilled water for 15 days. The plasma levels of big endothelin-1 (BET-1), creatine kinase-myocardial band (CK-MB), lactate dehydrogenase (LDH), aspartate aminotransferase (AST), and alanine aminotransferase (ALT), as well as the clinical status (heart rate and blood pressure), were assessed before and after treatment. Statistical analysis was performed with SPSS software using parametric and nonparametric ANOVA. Results: Mebudipine and amlodipine reversed the increased plasma BET-1 values in the treated animals when compared with the HF control group (0.103 and 0.112 vs 0.231 pg/mL, respectively). The increased plasma levels of AST, ALT, CK-MB, and LDH were also reversed in the HF animals that received mebudipine or amlodipine. Conclusion: The administration of mebudipine to HF animals, akin to amlodipine, palliated the clinical and biochemical signs of the disease in the present study. The abstract was presented in the Iranian Congress of Physiology and Pharmacology as a poster and published in the Scientific Information Database as a supplement (2015; Vol 22).

Collection of biochemical samples with brain-wide electrophysiological recordings from a freely moving rodent.

Bridging accumulating insights from microscopic and macroscopic studies in neuroscience research requires monitoring of neuronal population dynamics and quantifying specific molecules or genes from the brain of identical animals. To this end, by minimizing the size and weight of an electrode array, we developed a method that records local field potential signals of multiple brain regions from one side of the hemisphere in a freely moving rodent. At the same time, extracellular cerebrospinal fluid for biochemical assays or a small part of brain tissue samples for gene expression assays are collected from the other side of the hemisphere. This method allows ongoing stable recordings and sample collections for at least two months. The methodological concept is applicable to a wide range of biological reactions at various spatiotemporal scales, allowing us to integrate an idea of physiolomics into existing omics analyses, leading to a new combination of multi-omics approaches.

Isolation-induced ultrasonic vocalizations in pups: A comparison between Long-Evans, Sprague-Dawley, and Wistar rats.

Rat pup ultrasonic vocalizations (USV) are usually studied in outbred rats belonging to either Long-Evans, Sprague-Dawley, or Wistar stocks, but these were not compared so far. We therefore performed a stock comparison and analyzed USV of male pups (postnatal day 11) belonging to these three stocks. Pups of all three stocks showed substantial isolation-induced USV, but differed in various call features, like call numbers, peak frequency, and frequency modulation. Also, three different call types were identified by means of a quantitative approach based on peak frequency and frequency modulation, and it was found that their proportions differed between stocks. These results are discussed with respect to functional aspects of pup USV.

Sex differences in juvenile play behavior differ among rat strains.

Juvenile male rats frequently play more than female rats, but the presence of sex differences is affected by testing conditions and may also depend on the strain of rat. In this experiment, we tested play and defensive behaviors in male and female Long-Evans, Sprague-Dawley, and Wistar rats. When observed with a cage mate during the juvenile period, Long-Evans rats played more than Wistar animals, but there were no sex differences in any strain. When tested with an unfamiliar sibling (not seen since weaning), both Long-Evans and Wistar rats played more than Sprague-Dawley animals, and Long-Evans females played more than males. We did not observe any sex or strain differences in defensive behaviors. Our data indicate that there are strain differences in play behavior, and sex differences in play depend on both strain and context. Variation among strains may reflect underlying differences in anxiety, novelty seeking, and circadian rhythms.

Sexual Dimorphism of NADPH Oxidase/H2O2 System in Rat Thyroid Cells; Effect of Exogenous 17ß-Estradiol.

It has long been observed that females are more susceptible to thyroid diseases than males. Epidemiological and experimental data show that actions of hormonal factors-especially estrogens-may explain such disparity. However, the exact cause and mechanisms of this sexual dimorphism remain so far unknown. Therefore, we aimed at evaluating the effect of 17ß-estradiol on the redox balance in thyroids of male and female rats. Expression of nicotinamide adenine dinucleotide phosphate (NADPH) oxidases, i.e., dual oxidase 1 (DUOX1), dual oxidase 2 (DUOX2) and NADPH oxidase 4 (NOX4), and hydrogen peroxide (H2O2) levels were evaluated in the primary cell cultures derived from thyroid glands of adult male or female Wistar rats. The measurement was made before and after treatment with 17ß-estradiol alone or with addition of one of its receptor antagonists. We found that under basal conditions female thyroid cells are exposed to higher concentrations of H2O2, most likely due to NOX/DUOX enzymes activity. Additionally, exogenous 17ß-estradiol stimulated NOX/DUOX expression as well as H2O2 production, and this effect was mainly mediated through ERα. In conclusion, oxidative processes may constitute mechanisms responsible for sexual dimorphism of thyroid diseases. Exogenous 17ß-estradiol may play a crucial pathogenic role in thyroid diseases via oxidative mechanisms, however without any gender differences.

Role of Nitric Oxide Pathway in Development and Progression of Chronic Kidney Disease in Rats Sensitive and Resistant to its Occurrence in an Experimental Model of 5/6 Nephrectomy.

BACKGROUND Understanding the mechanisms conditioning development of chronic kidney disease (CKD) is still a challenge. The aim of this study was to evaluate the activity of the intrarenal nitric oxide (NO) pathway in the context of sensitivity or resistance of different animal strains to the development and degree of renal failure. MATERIAL AND METHODS Two rat strains were used: Wistar (WR) and Sprague-Dawley rats (SDR) in a model of CKD - 5/6 nephrectomy. We assessed parameters of renal failure and expression of nitric oxide synthase (NOS) isoforms in renal cortex and medulla. RESULTS We did not observe renal failure in WR, and CKD developed in SDR with increase of creatinine and urea concentration as well as decrease of diuresis and glomerular filtration. In the renal cortex, baseline expression of NOS2 was higher in WR than in SDR. 5/6 nephrectomy resulted in reduction of NOS2 in both strains and NOS3 in WR. In the renal medulla, baseline NOS2 expression was higher in SDR, and nephrectomy resulted in its decrease only in SDR. Although baseline NOS3 expression was higher in SDR, the NOS3 expression after nephrectomy was higher in WR rats. CONCLUSIONS In model of CKD - 5/6 nephrectomy, SDR proved to be sensitive and WR resistant to development of CKD. The intrarenal activity of the nitric oxide pathway was the factor that differentiated both strains. This mechanism may be responsible for insensitivity of WR to development of renal failure in this model of CKD.

Comparative ventilatory strategies of acclimated rats and burrowing plateau pika (Ochotona curzoniae) in response to hypoxic-hypercapnia.

The objective of this study was to compare the different ventilatory strategies that help in coping with hypoxic-hypercapnia environment among two species: use acclimated rats and plateau pikas (Ochotona curzoniae) that live in Tibetan plateaus, and have been well adjusted to high altitude. Arterial blood samples taken at 4100 m of elevation in acclimatized rats and adapted pikas revealed inter-species differences with lower hemoglobin and hematocrit and higher blood pH in pikas. A linear and significant increase in minute ventilation was observed in pikas, which help them to cope with hypoxic-hypercapnia. Pikas also displayed a high inspiratory drive and an invariant respiratory timing regardless of the conditions. Biochemical analysis revealed that N-methyl-D-aspartate receptor (NMDA) receptor gene and nNOS gene are highly conserved between rats and pikas, however pikas have higher expression of NMDA receptors and nNOS compared to rats at the brainstem level. Taken together, these results suggest that pikas have developed a specific ventilatory pattern supported by a modification of the NMDA/NO ventilatory central pathways to survive in extreme conditions imposed on the Tibetan plateaus. These physiological adaptive strategies help in maintaining a better blood oxygenation despite high CO2 concentration in burrows at high altitude.

Rat aversion to isoflurane versus carbon dioxide.

Some experts suggest that sedation of laboratory rodents with isoflurane before euthanasia with carbon dioxide (CO(2)) is a humane alternative to euthanasia with CO(2) alone, but little research has compared aversion with these agents. Albino rats were tested in a light-dark box where they had the choice between remaining in a dark compartment filling with isoflurane or CO(2), or escaping to a lit compartment. Experiment 1 validated the procedure by confirming that rats responded to agent and light intensity. In experiment 2, 9/16 and 0/16 rats remained in the dark compartment until recumbent when initially exposed to isoflurane and CO(2), respectively. In experiment 3, more rats remained in the dark compartment until recumbent during initial (10/16) versus re-exposure (1/16) to isoflurane. These results indicate that initial exposure to CO(2) is more aversive than isoflurane, and that re-exposure to isoflurane is more aversive than initial exposure. We conclude that sedation with isoflurane is a refinement over euthanasia with CO(2) alone for rats that have not been previously exposed to inhalant anaesthetics.

Findings in Historical Control Harlan RCCHanTM: WIST Rats from 4-, 13-, 26-Week Studies.

Vehicle control Harlan RCCHan™: WIST rats were examined to provide control data for subsequent studies. Sixty male and 60 female rats were sacrificed after 4, 13, and 26 weeks (360 animals total) of daily oral gavage dosing with reverse osmosis water. At necropsy, body weights, organ weights, and macroscopic findings were recorded, and tissues were collected for histopathology. Mean terminal body and organ weight data demonstrated expected age-related trends. Macroscopic findings occurred sporadically, generally at singular or at very low incidence, and with no observable age-related trend. The most frequent observation was discoloration of the stomach mucosa. Neoplastic microscopic findings were uncommon (one endometrial stromal polyp; one hepatocellular adenoma; one C-cell adenoma; and one sarcoma, NOS). The most common and/or notable nonneoplastic microscopic findings included basophilic tubules and mononuclear cell infiltration in the kidney, macrophage infiltration in pulmonary alveoli, and mononuclear infiltration in the liver of males and females, and myocardial degeneration/necrosis and mononuclear cell infiltration in the heart of males. Female reproductive tracts were staged to establish a representative baseline distribution. Diestrus, proestrus, estrus, and metestrus were diagnosed 45.8%, 11.9%, 30.5%, and 11.9%, respectively, at 4 weeks and 27.6%, 13.8%, 50.0%, and 8.6%, respectively, at 13 weeks.

D-Aspartic acid is a novel endogenous neurotransmitter.

D-aspartic acid (D-Asp) is present in invertebrate and vertebrate neuroendocrine tissues, where it carries out important physiological functions and is implicated in nervous system development. We show here that D-Asp is a novel endogenous neurotransmitter in two distantly related animals, a mammal (Rattus norvegicus) and a mollusk (Loligo vulgaris). Our main findings demonstrate that D-Asp is present in high concentrations in the synaptic vesicles of axon terminals; synthesis for this amino acid occurs in neurons by conversion of L-Asp to D-Asp via D-aspartate racemase; depolarization of nerve endings with K(+) ions evokes an immediate release of D-Asp in a Ca(2+) dependent manner; specific receptors for D-Asp occur at the postsynaptic membrane, as demonstrated by binding assays and by the expansion of squid skin chromatophores; D-aspartate oxidase, the specific enzyme that oxidizes D-Asp, is present in the postsynaptic membranes; and stimulation of nerve endings with D-Asp triggers signal transduction by increasing the second messenger cAMP. Taken together, these data demonstrate that D-Asp fulfills all criteria necessary to be considered a novel endogenous neurotransmitter. Given its known role in neurogenesis, learning, and neuropathologies, our results have important implications for biomedical and clinical research.

Metabolic and cardiac autonomic effects of high-intensity resistance training protocol in Wistar rats.

The aim of this study was to assess the effects of metabolic and autonomic nervous control on high-intensity resistance training (HRT) as determined by pancreatic glucose sensitivity (GS), insulin sensitivity (IS), blood lactate ([La]), and heart rate variability (HRV) in rats. Thirty male, albino Wistar rats (292 ± 20 g) were divided into 3 groups: sedentary control (SC), low-resistance training (LRT), and HRT. The animals in the HRT group were submitted to a high-resistance protocol with a progressively increasing load relative to body weight until exhaustion, whereas the LRT group performed the same exercise regimen with no load progression. The program was conducted 3 times per week for 8 weeks. The [La], parameters related to the functionality of pancreatic tissue, and HRV were measured. There was a significant increase in peak [La] only in the HRT group, but there was a reduction in [La] when corrected to the maximal load in both trained groups (LRT and HRT, p < 0.05). Both trained groups exhibited an increase in IS; however, compared with SC and LRT, HRT demonstrated a significantly higher GS posttraining (p < 0.05). With respect to HRV, the low-frequency (LF) band, in milliseconds squared, reduced in both trained groups, but the high-frequency band, in milliseconds squared and nu, increased, and the LF in nu, decreased only in the HRT group (p < 0.05). The HRT protocol produced significant and beneficial metabolic and cardiac autonomic adaptations. These results provide evidence for the positive benefits of HRT in counteracting metabolic and cardiovascular dysfunction.

Plasticity of olfactory bulb inputs mediated by dendritic NMDA-spikes in rodent piriform cortex.

The piriform cortex (PCx) is essential for learning of odor information. The current view postulates that odor learning in the PCx is mainly due to plasticity in intracortical (IC) synapses, while odor information from the olfactory bulb carried via the lateral olfactory tract (LOT) is 'hardwired.' Here, we revisit this notion by studying location- and pathway-dependent plasticity rules. We find that in contrast to the prevailing view, synaptic and optogenetically activated LOT synapses undergo strong and robust long-term potentiation (LTP) mediated by only a few local NMDA-spikes delivered at theta frequency, while global spike timing-dependent plasticity (STDP) protocols failed to induce LTP in these distal synapses. In contrast, IC synapses in apical and basal dendrites undergo plasticity with both NMDA-spikes and STDP protocols but to a smaller extent compared with LOT synapses. These results are consistent with a self-potentiating mechanism of odor information via NMDA-spikes that can form branch-specific memory traces of odors that can further associate with contextual IC information via STDP mechanisms to provide cognitive and emotional value to odors.

The Effect of Noise in the Intensive Care Unit on the Oxidative Stress Response in Rats.

OBJECTIVE: The objective of this study was to determine the effect of noise in the intensive care unit (ICU) on oxidative stress in a rat model. METHOD: This study had both a descriptive and a randomized controlled experimental stage. In the descriptive stage, to create a laboratory model of noise in the ICU, the noise level was measured for 24 hr on a randomly selected day in a surgical ICU, and voice recording was performed using a sound recording device. In the experimental stage, 30 male Wistar albino rats were randomly divided into 5 groups: a control group and groups exposed to the recording of the noise from the ICU for 24, 48, 72, and 168 hr. RESULTS: The noise level in the ICU was higher than the levels recommended for hospitals. Plasma corticosterone levels of the rats in the group exposed to the ICU noise for 168 hr were significantly higher than those of the control group. Plasma total protein values were significantly reduced in the rats exposed to 48, 72, and 168 hr of ICU noise compared to those of the control group. Superoxide dismutase activity was significantly decreased and malondialdehyde levels significantly increased in serum, spleen, and brain tissues as the duration of noise exposure increased. CONCLUSION: Findings reveal that rats experienced increasing levels of stress and oxidative stress as time exposed to the ICU noise increased. These results suggest that interventions to reduce noise in the ICU may be warranted.

Inter-laboratory control data for reproductive endpoints required in the OPPTS 870.3800/OECD 416 reproduction and fertility test.

BACKGROUND: The U.S. EPA revised the Reproduction and Fertility Effects Test Guideline (OPPTS 870.3800/OECD 416) in 1998, adding numerous endpoints in an effort to incorporate new methodologies, improve the sensitivity for detecting reproductive toxicants, and more efficiently utilize study animals. Many of these new endpoints have not been used in regulatory reproductive toxicology studies prior to their inclusion in the test guidelines; thus, the Health and Environmental Sciences Institute (HESI) of the International Life Sciences Institute (ILSI) initiated the Reproductive Endpoints Project to examine the utility of these new endpoints. METHODS: This report provides a retrospective analysis of 43 multi-generation studies (16 in Wistar rats, 27 in Sprague-Dawley rats) conducted according to the latest version of the test guidelines. It focuses on vehicle (negative) control values (means and ranges) for the various endpoints to examine inter-laboratory variability. RESULTS: Based on the compiled data, the most variable endpoints across laboratories and their associated coefficients of variation (CV) for each generation were: percent abnormal sperm (166-205%), testicular spermatid concentration (126-147%), postimplantation loss (97-104%), primordial follicle counts (69%, only measured in P2 females), and epididymal sperm concentration (52-57%). Absolute and relative prostate and thymus weights, weanling uterine weights, and anogenital distance had CVs of 25-50%. Sources of variability included procedural differences between laboratories, inherent biological variability, and/or small sample sizes for some endpoints. CONCLUSIONS: These inter-laboratory control data provide a means for laboratories to review their performance on reproductive toxicity measures, and provide perspective for interpreting their own control data and data from treated animals.

Testing the potential of 50 kHz rat calls as a species-specific rat attractant.

The control of mammalian pests relies heavily on the use of pesticides that are often avoided and are not species-specific. These problems are particularly acute for pesticides used to control rats (Rattus spp.). The efficacy and targeting of control could be improved by attracting animals to control measures using species-specific cues. One cue that has the potential to attract rats is the 50 kHz calls they emit in positive social situations. Here we test the potential of these rat calls as a species-specific attractant by examining the response of laboratory rats (Rattus norvegicus; n = 48) and non-target bank voles (Myodes glareolus; n = 16) to 50 kHz calls from either sex in a compartmentalised laboratory arena. Sounds of rat movement and white noise acted as control treatments, with each sound tested against a silent control in the opposite side of the arena. When sound cues were played above an empty bait box, rats were attracted to spend time close to 50 kHz rat calls, climbing on top of boxes, regardless of the sex of subject or caller. When either 50 kHz rat calls or rat movement sounds were played inside an empty bait box, rats of both sexes spent 3-4 fold more time inside boxes and visited more frequently. Rats were not attracted by intermittent white noise. Bank voles were neither attracted to, nor avoided, 50 kHz rat calls played inside empty bait boxes. Our findings show that 50 kHz rat calls are an effective attractant for rats of both sexes under laboratory conditions, while not attracting non-target bank voles. These calls are strong candidates for providing a species-specific lure that may be attractive even in the absence of food bait, but further trials will be needed to assess their efficacy under field conditions.

Relationship between motor function and ultrasonic vocalizations induced by maternal separation in rat pups.

When rat pups are isolated from their mothers, they emit ultrasonic vocalizations (USVs). Although previous studies have reported that USVs are related to anxiety, others have reported that they are related to simple, nonemotional factors, such as physiological reactions to coldness. In this study, we examined the influence of three maternal separations on rat pups. The number of USVs during 5 min of USV test under maternal separation, latency in the righting reflex as motor function, and body temperature were recorded twice (the first and second tests) before and after the pups were put in various environments for 10 min. The environments were no maternal separation (Control: CON), maternal separation with littermates (LMS), and single maternal separation with a heater (SMS). In the second test, the SMS pups had fewer USVs, a lower body temperature, and a more rapid righting reflex than the CON and LMS pups. In addition, there was no strong correlation between USVs and righting reflex. As a result, pups undergoing 10 min of SMS while being kept warm by the heater showed rapid righting reflex. Thus, by a single maternal separation, the number of USVs decreased but the decrease was unrelated to decrease in motor function.

Physiological changes of growth hormone during lactation in pup rats artificially reared.

During the lactation period, rat pups are fed by the dam, and the patterns of mother-pup interaction change during this period. Additionally, there are changes in feeding; first, mother´s milk is the only food needed for sustenance, and later, it is combined with solid food and water. GH serum concentrations depend on both maternal-pup interaction and energy metabolism. In the artificial rearing (AR) procedure, pups are deprived of mother-pup interaction, and the feeding pattern is controlled. This rearing paradigm has been used in rats to analyze the effects of maternal deprivation on social behavior. In the present study, we analyzed the variation in GH, acylated ghrelin and IGF-1 serum concentrations throughout the lactation period in AR pups. At pnd7, the maternal rearing (MR) pups responded to a 4 h fast with a drop in GH serum concentration, which is a well-known response to maternal deprivation. GH serum levels in the AR pups did not change, suggesting an adaptation phenomenon. A dopamine inhibitory effect of GH secretion was observed in pnd7 cultured somatotropes, suggesting dopamine regulation of GH secretion at this age. Acylated ghrelin serum levels in the AR pups showed an inverted pattern compared to that in the MR pups, which was related to the artificial feeding pattern. IGF-1 serum levels were lower in the AR pups than in MR pups, which was associated with hepatic GH resistance and with low Igf1 mRNA expression at pnd7. Interestingly, at pnd14, both pup groups showed high hepatic Igf1 mRNA expression but low IGF-1 serum levels, and this was inverted at pnd21. However, serum glucose levels were lower in the AR pups at pnd14 but reached the same levels as the MR pups at pnd21. Moreover, hepatomegaly and higher hepatic GH-receptor levels were observed in the AR pups at pnd21, which was in agreement with an absence of a solid food meal. During AR, the pups lost the maternal interaction-stimulated GH secretion, which correlated with lower IGF-1 serum levels during the first week of postnatal development. Later, the AR pups exhibited hepatic responses, in order to satisfy the metabolic demand for the normal weaning, with low carbohydrates levels in their meal.

Efficient derivation of knock-out and knock-in rats using embryos obtained by in vitro fertilization.

Rats are effective model animals and have contributed to the development of human medicine and basic research. However, the application of reproductive engineering techniques to rats is not as advanced compared with mice, and genome editing in rats has not been achieved using embryos obtained by in vitro fertilization (IVF). In this study, we conducted superovulation, IVF, and knock out and knock in using IVF rat embryos. We found that superovulation effectively occurred in the synchronized oestrus cycle and with anti-inhibin antiserum treatment in immature rats, including the Brown Norway rat, which is a very difficult rat strain to superovulate. Next, we collected superovulated oocytes under anaesthesia, and offspring derived from IVF embryos were obtained from all of the rat strains that we examined. When the tyrosinase gene was targeted by electroporation in these embryos, both alleles were disrupted with 100% efficiency. Furthermore, we conducted long DNA fragment knock in using adeno-associated virus and found that the knock-in litter was obtained with high efficiency (33.3-47.4%). Thus, in this study, we developed methods to allow the simple and efficient production of model rats.

Comparison of visual function in pigmented and albino rats by electroretinography and visual evoked potentials.

PURPOSE: To check differences in visual function between Wistar (albino) and Long-Evans (pigmented) rats. METHODS: The animals were born in our facilities and reared under identical light conditions avoiding bright light. Visual electrophysiology was performed at the ages of 1.5, 4, 7 and 10 months (electroretinography, ERG) and at 1.5 and 7 months (visual evoked potentials, VEP). RESULTS: ERG measurements showed that: 1) The amplitudes of both scotopic and photopic b-waves were markedly larger in Long-Evans rats than in Wistar rats, and also the amplitudes of scotopic oscillatory potentials and photopic 30 Hz Flicker amplitudes, 2) scotopic a-wave amplitudes were larger in Wistar rats at low light intensities, whereas they were smaller in bright light, 3) both a-wave and b-wave latencies were shorter in Wistar rats, 4) the maximum response Rm(P3) was larger in Long-Evans rats, 5) the sensitivity parameter S was larger in Wistar rats, and 6) the post-receptoral response of cones was smaller in Wistar rats. In the VEP measurements, amplitudes of both photopic and scotopic visual evoked potentials of Long-Evans rats were only slightly larger than those of Wistar rats. CONCLUSIONS: ERG b-wave amplitudes are markedly decreased in Wistar rats, which requires further investigation. As the b/a and OP/a ratios were also decreased in Wistar rats, it can be suggested that post-receptoral processing, in particular, is impaired in albino animals.

Strain differences in toxic effects of long-lasting isoflurane anaesthesia between Wistar rats and Sprague Dawley rats.

We investigated if long-lasting (5 h) anaesthesia with isoflurane has different pharmacological effects in two different rat strains: Wistar and Sprague Dawley. The mean blood pressure was 34% higher in Sprague Dawley rats as compared to the Wistar rats (p = 0.04). In Wistar rats, the pH value decreased to 7.1, lactate increased by 53%, creatinine increased 2.7-fold, alanine amino transferase and aspartate amino transferase increased more than 4-fold and lactate dehydrogenase increased 9-fold (p < 0.05). There were no changes in laboratory parameters in Sprague Dawley rats. This indicates that the Wistar rats were more sensitive to a 5 h anaesthesia with isoflurane after a premedication with ketamin/xylazine in the described study design.