Resistant Rats Research Articles

Voluntary food restriction does not affect circulating corticosterone in obesity-prone or -resistant male and female rats

Food restriction in rodents can increase circulating corticosterone, which reflects activation of physiological stress responses. These responses affect a myriad of behaviors and physiological processes and can increase the risk of obesity. Most of these studies have used experimenter-imposed restriction. However, rats will voluntarily restrict their food intake if they are returned to chow after a period of access to sugary, fatty “junk food” (JF) diet. Here we examine the effects of voluntary food restriction in obesity-prone and -resistant rats on circulating corticosterone concentrations and determine whether corticosterone release in response to acute stress differs in groups with a history of JF consumption.

Reversal of insulin resistance to combat type 2 diabetes mellitus by newer thiazolidinedione's in fructose induced insulin resistant rats

In our pursuit of discovering new antidiabetic agents to manage type 2 diabetes mellitus (T2DM), our approach aimed to identify the bioactive feature/pharmacophore responsible for PPAR-γ expression, as it is accountable for the glucose homeostasis and lipid metabolism. This was achieved by pharmacophore model generation, screening of rationally designed newer thiazolidinedione's library, identifying synthesizing and characterizing the top ten molecules (5a-5j) for their (Invitro &invivo) antidiabetic activity. Preliminary screening of all the ligands by Invitro glucose uptake assay in L6 myotubes (skeletal muscle cell line of rats) revealed compound 5b and 5f stimulated the glucose uptake with 79.29 ± 1.02 % and 74.58 ± 1.02 % respectively compared to pioglitazone with 82.36 ± 0.98 %. This was validated by PPAR-γ TF expression assay, which highlighted a dose dependent increase in transactivation of PPAR-γ. These compounds 5b and 5f were evaluated in fructose induced insulin resistance rat model. Where the treatment with 5b and 5f markedly increased the exogenous clearance of glucose and exogenous insulin via OGTT and ITT respectively, also improved the glucose utilization by significantly increasing content of glycogen and uptake of glucose in rat hemidiaphragm and reversed insulin resistance. Likewise a significant decreased in the VLDL and triglyceride levels was seen in 5b and 5f treated groups compared to insulin resistant (IR) group. It improved glycogenesis by catabolism of glucose and maintained glycaemic control. Similarly it had marked action on enzymatic oxidative biomarkers. Compound 5b displayed better, improved T1/2 (half-life) of 4.21 h and Kel (elimination constant) of 0.381 was noticed in comparison to compound 5f indicating the pharmacokinetic profile. Insilico studies like DFT calculations refined the geometry of 5b and 5f ligands, docking and molecular simulation provided the insights in binding affinity, dynamic behaviour and stability of ligands in PPAR-γ ligand binding domain. MM/GBSA provided the energetics of 5b and 5f in binding pocket. Finally network pharmacology identified ADIPOQ (adiponectin), NR1C3 (PPAR-γ), SLC2A4 (GLUT4), and LEP (leptin) proteins associate with compound 5b and 5f and enriched in Adipocytokine pathway, and PPAR-γ signaling pathway.

Gene expression of Decompression Sickness-resistant rats through a miRnome/transcriptome crossed approach

Susceptibility to decompression sickness (DCS) is characterized by a wide inter-individual variability, the origins of which are still poorly understood. We selectively bred rats with at least a 3-fold greater resistance to DCS than standard rats after 6 generations. In order to better understand DCS mechanisms, we compared the static genome expression of these resistant rats from the 10th generation to their counterparts of the initial non-resistant Wistar strain, by a microarray transcriptomic approach coupled and crossed with a PCR plates miRnome study. Thus, we identified differentially expressed genes on selected males and females, as well as gender differences in those genes, and we crossed these transcripts with the respective targets of the differentially expressed microRNAs. Our results highlight pathways involved in inflammatory responses, circadian clock, cell signaling and motricity, phagocytosis or apoptosis, and they confirm the importance of inflammation in DCS pathophysiology.

Monoclonal Antibody chP3R99 Reduces Subendothelial Retention of Atherogenic Lipoproteins in Insulin-Resistant Rats: Acute Treatment Versus Long-Term Protection as an Idiotypic Vaccine for Atherosclerosis.

Atherosclerosis is triggered by the retention of apolipoprotein B-containing lipoproteins by proteoglycans. In addition to low-density lipoprotein, remnant lipoproteins have emerged as pivotal contributors to this pathology, particularly in the context of insulin resistance and diabetes. We have previously reported antiatherogenic properties of a monoclonal antibody (chP3R99) that recognizes sulfated glycosaminoglycans on arterial proteoglycans. Solid-phase assays demonstrated that chP3R99 effectively blocked >50% lipoprotein binding to chondroitin sulfate and vascular extracellular matrix invitro. The preperfusion of chP3R99 (competitive effect) resulted in specific antibody-arterial accumulation and reduced fluorescent lipoprotein retention by ~60% in insulin resistant JCR:LA-cp rats. This competitive reduction was dose dependent (25-250 μg/mL), effectively decreasing deposition of cholesterol associated with lipoproteins. In a 5-week vaccination study in insulin resistant rats with (200 μg subcutaneously, once a week), chP3R99 reduced arterial lipoprotein retention, and was associated with the production of antichondroitin sulfate antibodies (Ab3) able to accumulate in the arteries (dot-blot). Neither the intravenous inoculation of chP3R99 (4.5 mg/kg), nor the immunization with this antibody displayed adverse effects on lipid or glucose metabolism, insulin resistance, liver function, blood cell indices, or inflammation pathways in JCR:LA-cp rats. Both acute (passive) and long-term administration (idiotypic cascade) of chP3R99 antibody reduced low-density lipoprotein and remnant lipoprotein interaction with proteoglycans in an insulin-resistant setting. These findings support the innovative approach of targeting proatherogenic lipoprotein retention by chP3R99 as a passive therapy or as an idiotypic vaccine for atherosclerosis.

Association between rat decompression sickness resistance, transthyretin single nucleotide polymorphism, and expression: A pilot study.

Decompression sickness (DCS) is a systemic syndrome that can occur after an environmental pressure reduction. Previously, we showed that the plasmatic tetrameric form of transthyretin (TTR) nearly disappeared in rats suffering DCS but not in asymptomatic ones. In this pilot study, we assessed whether the resistance to DCS could be associated with polymorphism of the gene of TTR. For this study, Sanger sequencing was performed on purified PCR products from the liver of 14-week-old male and female standard and DCS-resistant rats (n = 5 per group). Hepatic TTR mRNA expression was assessed by RT-qPCR in 18-19 week-old male and female standard and resistant rats (n = 6 per group). There is a synonymous single nucleotide polymorphism (SNP) on the third base of codon 46 (c.138 C > T). The thymine allele was present in 90% and 100% of males and females standard, respectively. However, this allele is present in only 30% of DCS-resistant males and females (p = 0.0002301). In the liver, there is a significant effect of the resistance to DCS (p = 0.043) and sex (p = 0.047) on TTR expression. Levels of TTR mRNA were lower in DCS-resistant animals. To conclude, DCS resistance might be associated with a SNP and a lower expression of TTR.

Food Cue Reactivity Meets the Reinforcer Pathology Model: Behavioral Economic Measures of Cue-Induced Changes in Food Reinforcer Efficacy

Food cue reactivity, or behavioral sensitivity to conditioned food cues, is an eating pattern observed in those with obesity and binge-eating disorder. The reinforcer pathology model, which characterizes overconsumption of a reinforcer such as food may be relevant to food cue reactivity, especially in those with obesity and binge-eating disorder. The reinforcer pathology model posits that steep delay discounting (DD) and demand elasticity are processes involved in the overconsumption of food. Two of our recent studies examine the extent to which reactivity to conditioned food cues may be involved in food reinforcer pathologies. First, food cues were conditioned with Oreo cookies with binge-eating prone (BEP) and binge-eating resistant (BER) rats. Delay discounting was compared before and after conditioning. Food cues induced steeper DD for rats, though BEP rats showed some evidence for greater sensitivity to this effect than BER rats, albeit this difference was not significant. Second, healthy-weight humans and humans with overweight/obese BMI underwent conditioning of visual cues paired with M&M candies. After acquisition, cues induced greater demand intensity and inelasticity for food compared to baseline. Participants with overweight/obese BMI, compared to controls, also showed some evidence for greater sensitivity to this change ininelasticity compared to healthy-weight participants, but this difference was also not significant. Food cues, then, may induce changes in DD and economic demand, supporting the relevance of reinforcer pathologies.

Exogenous T4 Increased Cardiac Thioredoxin Protein Expression in Insulin Resistant OLETF Rats

Thioredoxin (Trx) is a protein that regulates cellular signaling and antioxidant defense. Cellular antioxidant mechanisms are designed to protect cells from oxidative damage caused by inappropriately elevated reactive oxygen species (ROS) and other free radicals. Oxidative damage can lead to cellular damage and impaired function. Diabetic cardiomyopathy (CDM) is associated with impaired redox balance and oxidative injury. Altered fuel metabolism is a key feature of CDM characterized by an imbalance in fatty acid and glucose utilization, favoring fatty acid overutilization. Excessive fatty acid beta-oxidation in the mitochondria leads to generation of free radicals including superoxide, which can contribute to oxidative injury. Thyroid hormones regulate substrate utilization and their dysregulation may contribute to oxidative injury. Therefore, we hypothesized that exogenous thyroxine (exoT4) upregulates Trx protein expression and ameliorate oxidative injury in the hearts of insulin resistant rats. Insulin resistant, Otsuka Long Evans Tokushima Fatty (OLETF) rats were used to assess the effects of exoT4 on heart Trx, oxidative damage, and glucose metabolism. Rats were assigned to four groups: (1) lean, control Long Evans Tokushima Otsuka (LETO; n=6), (2) LETO + T4 (8 μg/100g BM/d × 5 wks; n=7), (3) untreated, insulin resistant OLETF (n=6), and (4) OLETF + T4 (n=7). OLETF group displayed a 31% decreased in TRX compared to Leto. T4 increased Trx protein expression by 88% in OLTEF compared to untreated OLETF. This data suggests that exoT4 has the potential to upregulate cardiac Trx-dependent antioxidant mechanisms in insulin resistant OLETF rats to ameliorate oxidative injury and improve cardiac glucose metabolism. This work is supported by the American Heart Association and the Undergraduate Research Opportunity Center (UROC). This is the full abstract presented at the American Physiology Summit 2024 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.

Exogenous thyroxine increases cardiac GLUT4 translocation in insulin resistant OLETF rats

During insulin resistance, the heart undergoes a metabolic shift in which fatty acids (FA) account for roughly about 99% of the ATP production. This metabolic shift is indicative of impaired glucose metabolism. A shift in FA metabolism with impaired glucose tolerance can increase reactive oxygen species (ROS), lipotoxicity, and mitochondrial dysfunction, ultimately leading to cardiomyopathy. Thyroid hormones (TH) may improve the glucose intolerance by increasing glucose reabsorption and metabolism in peripheral tissues, but little is known on its effects on cardiac tissue during insulin resistance. In the present study, insulin resistant Otsuka Long Evans Tokushima Fatty (OLETF) rats were used to assess the effects of exogenous thyroxine (T4) on glucose metabolism in cardiac tissue. Rats were assigned to four groups: (1) lean, Long Evans Tokushima Otsuka (LETO; n=6), (2) LETO + T4 (8 μg/100 g BM/d × 5 wks; n = 7), (3) untreated OLETF (n = 6), and (4) OLETF + T4 (8 μg/100 g BM/d × 5 wks; n = 7). T4 increased GLUT4 gene expression by 85% in OLETF and increased GLUT4 protein translocation to the membrane by 294%. Additionally, T4 increased p-AS160 by 285%, phosphofructokinase-1 (PFK-1) mRNA, the rate limiting step in glycolysis, by 98% and hexokinase II by 64% in OLETF. T4 decreased both CPT2 mRNA and protein expression in OLETF. The results suggest that exogenous T4 has the potential to increase glucose uptake and metabolism while simultaneously reducing fatty acid transport in the heart of insulin resistant rats. Thus, L-thyroxine may have therapeutic value to help correct the impaired substrate metabolism associated with diabetic cardiomyopathy.

Activation of mitoKATP channels induces penile vasodilation and inhibits mitochondrial respiration and ROS production: Role of NO

ObjectiveMitochondrial ATP-sensitive K+ (mitoKATP) channels are involved in neuronal and cardiac protection from ischemia and oxidative stress. Penile erection is a neurovascular event mediated by relaxation of the erectile tissue via nitric oxide (NO) released from nerves and endothelium. In the present study, we investigated whether mitoKATP channels play a role in the control of penile vascular tone and mitochondrial dynamics, and the involvement of NO. MethodsThe effect of the selective mitoKATP activator BMS191095 was examined on vascular tone, on mitochondrial bioenergetics by real-time measurements with Agilent Seahorse and on ROS production by MitoSOX fluorescence in freshly isolated microarteries. ResultsBMS191095 and diazoxide relaxed penile arteries, BMS191095 being one order of magnitude more potent. BMS191095-induced relaxations were reduced by mechanical endothelium removal and by inhibitors of the nitric oxide synthase (NOS) and PI3K enzymes. The NO-dependent component of the relaxation to BMS191095 was impaired in penile arteries from insulin resistant obese rats. The blockers of mitoKATP channel 5-HD, sarcolemma KATP (sarcKATP) channel glibenclamide, and large conductance Ca2+-activated K+ (BKCa) channel iberiotoxin, inhibited relaxations to BMS191095 and to the NO donor SNAP. BMS191095 reduced the mitochondrial bioenergetic profile of penile arteries and attenuated mitochondrial ROS production. Blockade of endogenous NO impaired and exogenous NO mimicked, respectively, the inhibitory effects of BMS191095 on basal respiration and oxygen consumed for ATP synthesis. Exogenous NO exhibited dual inhibitory/stimulatory effects on mitochondrial respiration. ConclusionsThese results demonstrate that selective activation of mitoKATP channels causes penile vasodilation, attenuates ROS production and inhibits mitochondrial respiration in part by releasing endothelial NO. These mechanisms couple blood flow and metabolism in penile arterial wall and suggest that activation of vascular mitoKATP channels may protect erectile tissue against ischemic injury.

Distribution of anticoagulant resistance in the brown rat in Belgium

Anticoagulant resistance is known as one of the major factors interfering with rodent control. Within this context we investigated the distribution of anticoagulant resistance in Flanders, northern Belgium. From 2003 to 2005, we tested 691 rats from different locations with blood clotting response tests for their susceptibility to the anticoagulant compounds warfarin, bromadiolone and difenacoum. Of these, 119 were also screened for a mutation in the VKORC1 gene that is suspected to be responsible for anticoagulant resistance. Warfarin resistant rats were found in the western and eastern parts of Flanders. The same distribution pattern was found for bromadiolone with the exception of the south-eastern area, where this form of resistance was largely absent. We detected difenacoum resistance in only six rats and did not observe any resistant rats in the central part of Flanders. Susceptible rats were found all over Flanders. Genetic analyses showed that anticoagulant
 resistance in Belgium was related to two different mutations in VKORC1, namely Y139F and L120Q. Our results indicate that rodent control should be regionally tailored to be most effective.

Sensitivity assessment of diphacinone by pharmacokinetic analysis in invasive black rats in the Bonin (Ogasawara) Archipelago, Japan

The Bonin Archipelago is a United Nations Educational, Scientific and Cultural Organization's World Natural Heritage Site in Japan with a unique ecosystem; however, the invasive rodents preying on endemic species have been a significant concern. The anticoagulant rodenticide, diphacinone, sprayed by the Ministry of the Environment, has succeeded; however, its repeated use leads to rodenticide resistance. This study evaluated the sensitivity by in vivo pharmacokinetics/pharmacodynamics (PK/PD) analysis and physiologically-based pharmacokinetic modeling to diphacinone in black rats (Rattus rattus) captured on the Bonin Archipelago in February 2022. The Bonin rats exhibited prolonged coagulation time after diphacinone administration. They recovered earlier than susceptible black rats, indicating that Bonin rats were less susceptible, though there were no genetic mutations in Vkorc1, the target enzyme of diphacinone. After the administration of diphacinone, hepatic expression levels of Fsp1, identified as the vitamin K reductase, was decreased, however, the Bonin rats exhibited the most minor suppression. The PK analysis showed that the excretion capacity of the Bonin rats was lower than that of the resistant black rats. In the PBPK modeling, the resistant black rats showed higher clearance than the Bonin and susceptible black rats due to high hepatic metabolic capacity. The Bonin rats demonstrated slow absorption and relatively low clearance. This study highlighted the reduced rodenticide-sensitive tendency of wild black rats in the Bonin Archipelago at an in vivo phenotype level. At the same time, they do not have known rodenticide resistance mechanisms, such as hepatic metabolic enhancement or Vkorc1 mutations. It is crucial to monitor the biological levels to evaluate rodenticide sensitivity accurately.

Cyclosorus Terminans Extract Alleviates Neuroinflammation in Insulin Resistant Rats.

High-fat diet consumption for an extended period causes obesity, systemic metabolic disturbance, and brain insulin resistance, resulting in neuroinflammation. Although the beneficial effect of Cyclosorus terminans extract on obesity-related insulin resistance has been demonstrated, little is known about how it affects neuroinflammation and brain insulin resistance in obese rats. Male Wistar rats were given either a normal diet (ND, n = 6) or a high-fat diet (HFD, n = 24) for a total of 14 weeks. At the beginning of the week, 13 rats in the ND group were given vehicle orally for 2 weeks, while rats on HFD diets were randomized to one of four groups and given either vehicle, 100 mg/kg/day of Cyclosorus terminans extract, 200 mg/kg/day of Cyclosorus terminans extract, or 20 mg/kg/day of pioglitazone orally for 2 weeks. After the experimental period, blood and brain samples were taken to assess metabolic and brain parameters. HFD-fed rats had obesity, systemic and brain insulin resistance, brain inflammation, microglial and astrocyte hyperactivity, and brain necroptosis. Treatment with 200 mg/kg/day of Cyclosorus terminans extract and pioglitazone equally attenuated obesity, insulin resistance, brain insulin dysfunction, and neuroinflammation in insulin resistant rats. Our findings suggest that Cyclosorus terminans extract may hold promise as a therapeutic agent for insulin resistance and neuroinflammation in obese conditions.

Influence of Stress Resistance on Myocardial Expression of the Pro-Autophagic Protein Beclin-1 After Cardiac Contusion in Experimental Setting

Objective. Evaluation of myocardial expression of the pro-autophagic protein Beclin-1 after cardiac contusion in experimental animals with different stress resistance.Materials and methods. The study included 68 white mongrel male rats weighing 250–300 g. After ranking for extreme variants of stress resistance, moderately stress-resistant rats (N=36) were excluded from the study. The remaining animals were split into the control (N=16) and study (N=16) groups, each group composed of 8 high stress resistant and 8 low stress resistant rats. In the study group, 24 hours after inflicted cardiac contusion, 5×5 mm myocardial tissue specimens were sampled from the intraventricular septum, anterior walls of the left and right ventricles, histological sections were made, and a reaction with primary polyclonal Anti-Beclin-1 antibodies was performed. Beclin-1 expression was evaluated under the microscope.Results. Immunohistochemical evaluation revealed a statistically significant increase in Beclin-1 protein expression (P=0.0002) in the cytoplasm of cardiomyocytes in the study group vs the control group, regardless of animals’ baseline stress resistance. However, expression of Beclin-1 protein in the myocardium of highly stress-resistant rats (Me=4.3; LQ=4.0; HQ=4.3) was significantly higher versus low-resistant animals (Me=3.6; LQ=3.3; HQ=3.6) (P=0.0009).Conclusion. Increased expression of Beclin-1 protein in the post-traumatic period of experimental cardiac contusion indicates autophagic flux activation. Intensity of autophagy varied depending on the animal’s stress resistance.

Review of Decreased IL-6 Levels in SOPK-Insulin Resistant Rats fed a Low Carb High Protein Diet

Polycystic ovary syndrome with insulin resistance (SOPK-RI) is caused among endocrine disorders with an incidence rate of 5-10% that occurs in women of reproductive age. Characteristics of polycystic ovary syndrome are found anovulation, hyperandrogens, polycystic ovarii and impaired insulin sensitivity involving proinflammatory increases in several inflammatory cytokines including interleukin-6 (IL-6). Increased inflammatory cytokines in women with SOPK-RI found health problems there is fat accumulation in adipocyte tissue such as in an obese person, metabolic syndrome and type 2 diabetes. The purpose of this study was to determine the effect of a low-carbohydrate high-protein diet on changes in IL-6 levels in SOPK-RI model mice. Laboratory experimental research, namely post test only control group design. Consisting of three groups, namely, the negative control group (K-) as a normal group did not get treatment; The positive control group (K+) of the SOPK-insulin resistance model mice were given standard feed, and the treatment group (P) of the SOPK-insulin resistance model mice were given a low-carbohydrate high-protein diet (CTR) with a composition of 40% carbohydrate and 30% protein. The study lasted for 48 days, using serum ELISA examination on IL-6 levels. The average results of IL-6 levels, namely the K- = 0.358 group, K+ = 0.387 and P = 0.442 and obtained different IL-6 levels of significance value p = 0.002 with a low-carbohydrate high-protein diet in SOPK - RI rats showed a significant difference (p < 0.05) between the control group and the treatment group. Significantly reduced IL-6 levels in SOPK-RI model mice. A low-carb, high-protein diet may be considered as a dietary modality in women with SOPK-insulin resistance. Further research is needed to better determine the other modulators involved in this mechanism in more detail.

Study of fingolimod, nitric oxide inhibitor, and P-glycoprotein inhibitor in modulating the P-glycoprotein expression via an endothelin-sphingolipid pathway in an animal model of pharmacoresistant epilepsy.

The overexpression of P-glycoprotein (P-gp) contributes to drug resistance in patients with epilepsy, and the change of P-gp expression located at the blood-brain barrier alienates the anti-seizure effects of P-gp substrates. Thus, the present study explored the effect of fingolimod (FTY720) acting through an endothelin-sphingolipid pathway on P-gp-induced pentylenetetrazol (PTZ)-kindled phenobarbital (PB)-resistant rats. PTZ kindling (30 mg/kg; i.p.) and PB (40 mg/kg; orally) were used to develop an animal model of refractory epilepsy. The effect of Fingolimod on seizure score (Racine scale), plasma and brain levels of PB (high-performance liquid chromatography), and blood-brain barrier permeability (Evans blue dye) was determined. Further, Fingolimod's neuroprotective effect was determined by measuring the levels of various inflammatory cytokines, oxidative stress parameters, and neurotrophic factors in rat brain homogenate. The Fingolimod's effect on P-gp expression was estimated by reverse transcriptase-polymerase chain reaction and immunohistochemistry in rat brain. The H and E staining was done to determine the neuronal injury. Fingolimod significantly (P < 0.001) reduced the seizure score in a dose-dependent manner and alleviated the blood-brain barrier permeability. It decreased the P-gp expression, which further increased the brain PB concentration. Fingolimod significantly (P < 0.01) reduced oxidative stress as well as inflammation. Moreover, it attenuated the raised neuronal injury score in a resistant model of epilepsy. The modulation of the P-gp expression by Fingolimod improved drug delivery to the brain in an animal model of refractory epilepsy. Therefore, S1P signaling could serve as an additional therapeutic target to overcome refractoriness.

Genetically predisposed and resilient animal models of depression reveal divergent responses to early-life adversity.

Early-life adversity is one of the strongest predictors of childhood depression, that may be exacerbated by a genetic predisposition to develop depression. We therefore investigated the bio-behavioural effects of an early-life stressor in an accepted rodent model of depression. The Flinders sensitive (FSL) and resistant (FRL) line rats were subjected to an early-life stressor, whereafter their bio-behavioural response during pubertal onset was evaluated. Male and female pups were maternally separated for 3 h per day from postnatal day 02 (PND02) to 17, when they were also weaned. Control animals were left undisturbed, until weaning on PND21. Depressive-like behaviour was analysed on PND21 and reassessed on PND36. Hippocampal monoamine levels, markers of oxidative stress and metabolic markers implicating mitochondrial function were also measured. On PND21, the non-maternal separation and early weaning (non-MSEW) FSL rats spent 10% more time mobile than their FRL controls in the tail suspension test (TST) yet displayed increased depressive-like behaviour in the forced swim test (FST) on PND36. The depressive-like behaviour coincided with increased hippocampal norepinephrine levels, serotonin turnover and a dysfunctional redox state. Maternal separation and early weaning (MSEW) appeared to initially reduce early-life (PND21) depressive-like behaviour in the TST but then induced depressive-like behaviour on PND36, and increased norepinephrine levels more profoundly in the FRL rats. These findings highlight the need to further investigate the stress response pathway in these animals and that the absence or presence of genetic susceptibility may influence the presentation of early-life adversity effects.

VKORC1-based resistance to anticoagulant rodenticides widespread in Finnish house mice but not in brown rats

Anticoagulant rodenticide (AR) resistance is widespread across Europe, but its prevalence has not been previously studied in Finland. We sampled 48 house mice (Mus musculus) and 48 brown rat (Rattus norvegicus) in farms and urban locations from southern Finland and here we report the identification of Vkorc1 polymorphisms linked to AR resistance. Two thirds of mice carried Vkorc1 polymorphisms. The most common was the type Y139C (resistance to first-generation ARs, bromadiolone and difenacoum), whereas the type L128S (resistance to FGARs and bromadiolone) was much less abundant. Only two rats tested positive, both with a rare type R33P (resistance probably to warfarin). No clear patterns based on subregion, sex and urban/rural division were seen. We suggest that the type Y139C polymorphism should be taken into account when planning control actions targeting house mice in Finland. The low number of AR resistant rats found suggests a low prevalence of resistance in rats. To fully understand the state of AR resistance in Finland, additional studies on yellow-necked mouse (Apodemus flavicollis) and bank voles (Myodes glareolus) are needed.

Rapid, automated, and experimenter-free touchscreen testing reveals reciprocal interactions between cognitive flexibility and activity-based anorexia in female rats.

Anorexia nervosa has among the highest mortality rates of any psychiatric disorder and is characterized by cognitive inflexibility that persists after weight recovery and contributes to the chronic nature of the condition. What remains unknown is whether cognitive inflexibility predisposes individuals to anorexia nervosa, a question that is difficult to address in human studies. Our previous work using the most well-established animal model of anorexia nervosa, known as activity-based anorexia (ABA) identified a neurobiological link between cognitive inflexibility and susceptibility to pathological weight loss in female rats. However, testing flexible learning prior to exposure to ABA in the same animals has been thus far impossible due to the length of training required and the necessity of daily handling, which can itself influence the development of ABA. Here, we describe experiments that validate and optimize the first fully-automated and experimenter-free touchscreen cognitive testing system for rats and use this novel system to examine the reciprocal links between reversal learning (an assay of cognitive flexibility) and weight loss in the ABA model. First, we show substantially reduced testing time and increased throughput compared to conventional touchscreen testing methods because animals engage in test sessions at their own direction and can complete multiple sessions per day without experimenter involvement. We also show that, contrary to expectations, cognitive inflexibility measured by this reversal learning task does not predispose rats to pathological weight loss in ABA. Instead, rats that were predisposed to weight loss in ABA were more quickly able to learn this reversal task prior to ABA exposure. Intriguingly, we show reciprocal links between ABA exposure and cognitive flexibility, with ABA-exposed (but weight-recovered) rats performing much worse than ABA naïve rats on the reversal learning task, an impairment that did not occur to the same extent in rats exposed to food restriction conditions alone. On the other hand, animals that had been trained on reversal learning were better able to resist weight loss upon subsequent exposure to the ABA model. We also uncovered some stable behavioral differences between ABA susceptible versus resistant rats during touchscreen test sessions using machine learning tools that highlight possible predictors of anorectic phenotypes. These findings shed new light on the relationship between cognitive inflexibility and pathological weight loss and provide targets for future studies using the ABA model to investigate potential novel pharmacotherapies for anorexia nervosa.

Effects of rotating electric fields on liver biopolymers: an experimental study

BACKGROUND: According to the classic works of H. Selye, a variety of factors can impact humans and trigger a complex bodily response known as a stress reaction. This can lead to an imbalance in the body's regulatory physiological systems.&#x0D; AIM: To investigate the effects of a technogenic rotating electric field (REF) on the levels of carbohydrate-containing liver biopolymers in experimental animals.&#x0D; MATERIAL AND METHODS: A total of 54 rats were used in the experiment. The levels of sialic acids, mucoproteins, fucose, and -L-fucosidase were measured in the liver homogenate before the study, on the 10th and 20th day of the experiment. To ensure accurate results, the rats were first diagnosed using the open field method to determine their stress resistance levels. Based on the results, the rats were then divided into three groups: stress-resistant, stress-unstable, and ambivalent.&#x0D; RESULTS: By the 10th day of REF exposure, an increase in all the studied parameters in the liver homogenate in rats was observed indicating catabolic processes. Sialic acids concentration in stress-resistant, unstable and ambivalent rats increased by 14% (p=0.024), 29% (p=0.020) and 26% (p=0.021), respectively. Corresponding elevations of fucose concentration were 24% (p=0.019), 27% (p=0.019), 31% (p=0.019) while the activity of -L-fucosidase increased by 55% (p=0.024), in 63% (p=0.024) and 55% (p=0.011) in the abovementioned categories of rates. Mucoproteins concentrations increased by 58% (p=0.011) in stress-resistant, 76% (p=0.011) in stress-unstable and 65% (p=0.021) in stress-ambivalent rats. By the 20th day of the experiment, decomposition of carbohydrate-containing biopolymers slowed in all groups. When compared with the 10th day 10, sialic acids concentration decreased in stress resistant, unstable and ambivalent rats by 12% (p=0.041), 17% (p=0.021) and 20% (p=0.011), respectively. Corresponding decrease in of mucoproteins was 26% (p=0.011), 33% (p=0.024), and 32% (p=0.024). Fucose concentration increased by 34% (p=0.024) in stress-resistant, by 22% (p=0.024) in stress-unstable and by 28% (p=0.010) in stress-ambivalent rats. Correspondingly, -L-fucosidase activity increased by 15% (p=0.021), 46% (p=0.02) and 31% (p=0.011).&#x0D; CONCLUSION: The study's findings indicate that technogenic REF can alter the levels of carbohydrate-containing biopolymers in animal livers, leading to the activation of catabolic processes. The group of stress-unstable individuals exhibited the most significant catabolic processes. Our results may have implications for occupations exposures to REF.

Sex and Estrous Cycle Are Not Mediators of S-Ketamine's Rapid-Antidepressant Behavioral Effects in a Genetic Rat Model of Depression.

Recent preclinical and clinical studies have shed light on the possible impact of sex and estrous/menstrual cycle on ketamine's antidepressant action but with incongruous results. The preclinical studies that have shown the effects of ovarian sex hormones have not done so in animal models of depression. Thus, the aim of the present study is to scrutinize the acute behavioral responses to a subanesthetic dose of S-ketamine in males vs females and in different estrous phases in free-cycling females in a well-powered translational approach. We evaluated the behavioral sensitivity to 20 mg/kg S-ketamine (i.p.) in male and female Flinders Sensitive Line rats (FSLs) and their counterpart Flinders Resistant Line rats (FRLs) subjected to the open field and forced swim tests. Female rats were disaggregated into different estrous phases, and the behavioral outcomes were compared. Acute administration of S-ketamine had robust antidepressant-like effects in FSLs. Within our study power, we could not detect sex- or estrous cycle-specific different antidepressant-like responses to S-ketamine in FSLs. Fluctuations in the levels of ovarian sex hormones across different estrous cycles did not behaviorally affect S-ketamine's rapid-acting antidepressant mode of action. No sex-related or estrous cycle-related impact on behavioral despair was observed even among FRLs and saline-treated FSLs. We conclude that physiological oscillations of estrogen and progesterone levels neither amplify nor diminish the behavioral antidepressant-like effect of S-ketamine. In addition, fluctuations of ovarian sex hormones do not predispose female animals to exhibit enhanced or reduced depressive-like and anxiety-like behaviors.