Ritanserin reverses repeated methamphetamine-induced behavioral and neurochemical sensitization in mice

Chronic administration of methamphetamine (METH) elicits progressive enhancement of locomotor activity known as behavioral sensitization. We have recently shown that chronic METH enhanced METH challenge-induced increase in 5-HT levels in the prefrontal cortex and that 5-HT(1A) receptor activation attenuated this neurochemical sensitization as well as behavioral sensitization. This study examined whether the nonselective 5-HT(2) receptor antagonist, ritanserin affects METH-induced behavioral and neurochemical sensitization in mice. Ritanserin at doses of 1 and 3 mg/kg inhibited the development and expression of METH-induced behavioral sensitization in a dose-dependent manner. Furthermore, chronic administration of ritanserin for a week attenuated the maintenance of behavioral sensitization, indicating the improvement of established behavioral sensitization. Microdialysis analysis showed that chronic ritanserin inhibited the neurochemical sensitization that chronic METH enhanced METH challenge-induced increase in extracellular 5-HT levels in the prefrontal cortex. Furthermore, acute ritanserin inhibited METH challenge-induced increase in extracellular 5-HT but not DA levels in the prefrontal cortex. These results suggest that 5-HT(2) receptors are involved in METH-induced hyperactivity and behavioral sensitization in mice.     

Behavioral and neurochemical consequences of lipopolysaccharide in mice: anxiogenic-like effects

Systemic administration of lipopolysaccharide (LPS) induces sickness behaviors, as well as alterations of hypothalamic-pituitary-adrenal functioning commonly associated with stressors. In the present investigation, it was demonstrated that systemic LPS treatment induced a sickness-like behavioral profile (reduced active behaviors, soporific effects, piloerection, ptosis), which appeared to be dependent upon the novelty of the environmental context in which animals were tested. As well, LPS induced anxiogenic-like responses, including decreased time spent in the illuminated portion of a light-dark box, reduced open-arm entries in a plus-maze test, and decreased contact with a novel stimulus object in an open-field situation. The behavioral changes were accompanied by increased plasma ACTH and corticosterone levels. As well, LPS induced increased turnover of norepinephrine (NE), dopamine (DA) and serotonin (5-HT) in the paraventricular nucleus (PVN), median eminence plus arcuate nucleus, hippocampus, as well as NE turnover within the locus coeruleus and DA turnover within the nucleus accumbens. Although these neurochemical variations were reminiscent of those elicited by stressors, LPS was not particularly effective in modifying DA activity within the prefrontal cortex or NE within the amygdala, variations readily induced by stressors. Whether the LPS-induced anxiogenic-like responses were secondary to the illness engendered by the endotoxin remains to be determined. Nevertheless, it ought to be considered that bacterial endotoxin challenge, and the ensuing cytokine changes, may contribute to emotionality and perhaps even anxiety-related behavioral disturbances.     

Behavioral and neurochemical studies on brain aging in galanin overexpressing mice

To study possible involvement of galanin in brain aging quality, we have investigated behavioral, neurochemical and morphological parameters in aged mice overexpressing galanin under the platelet-derived growth factor B promoter (GalOE mice) compared to wild-type littermates (WT mice). The behavioral analysis in the forced swim test showed that old GalOE animals spent more time in immobility compared to WT. In the activity cage test, galanin overexpression counteracted the age-induced decrease in exploratory behavior. The neurochemical analysis showed a 30% decrease in noradrenaline overflow in the cerebral cortex of WT old mice that was not present in age-matched GalOE mice. Our results indicate that overexpression of galanin can influence several behavioral and neurochemical parameters in old mice.     

Altered neurotransmission in brains of autoimmune mice: pharmacological and neurochemical evidence

Depressive-like behavior is the most profound manifestation of autoimmunity-associated behavioral syndrome in lupus-prone MRL-lpr mice. This led to the hypothesis that chronic autoimmunity and inflammation alter the activity of central serotonergic and dopaminergic systems. Three drugs with a selective mode of action were used to probe the functional status of these two systems in vivo. The behavioral effects of single and repeated intraperitoneal (i.p.) injections of sertraline, quinpirole (QNP) and risperidone were measured in the forced swim and brief sucrose preference tests. In comparison to MRL +/+ controls, autoimmune MRL-lpr mice did not show a reduction in sucrose intake after the administration of sertraline. Acute injection of quinpirole increased floating more in the MRL-lpr than in the control group, while intermittent administration induced self-injurious behavior in both groups. Acute injection of risperidone significantly increased floating in MRL-lpr mice, while repeated administration abolished the difference between the substrains in sucrose intake. These discrepancies in responsiveness implied that the central neurotransmitter activity is dissimilar in the two MRL substrains. This notion was confirmed in a cohort of untreated MRL-lpr and MRL +/+ mice by comparing their neurotransmitter/metabolite levels in several brain regions. In particular, MRL-lpr brains showed increased dopamine (DA) levels in the paraventricular nucleus (PVN) and median eminence (ME), decreased concentrations of serotonin in the PVN and enhanced levels in the hippocampus, as well as decreased norepinephrine (NE) levels in the prefrontal cortex. Behavioral deficits correlated with the changes in PVN and median eminence. These results are consistent with the hypothesis that imbalanced neurotransmitter regulation of the hypothalamus-pituitary axis plays an important role in the etiology of behavioral dysfunction induced by systemic autoimmune disease.     

Behavioral and neurochemical effects of wild-type and mutated human alpha-synuclein in transgenic mice

Human alpha-synuclein (halpha-SYN) is implicated in the Parkinson's disease phenotype (PDP) based on a variety of studies in man, animal models, and in vitro studies. The normal function of halpha-SYN and the mechanism by which it contributes to the PDP remains unclear. We created transgenic mice expressing either wild-type (hwalpha-SYN) or a doubly mutated (hm2alpha-SYN) form of halpha-SYN under control of the 9-kb rat tyrosine hydroxylase promoter. These mice expressed halpha-SYN in cell bodies, axons, and terminals of the nigrostriatal system. The expression of halpha-SYN in nigrostriatal terminals produced effects in both constructs resulting in increased density of the dopamine transporter and enhanced toxicity to the neurotoxin MPTP. Expression of hm2alpha-SYN reduced locomotor responses to repeated doses of amphetamine and blocked the development of sensitization. Adult hwalpha-SYN-5 transgenic mice had unremarkable dopaminergic axons and terminals, normal age-related measures on two motor coordination screens, and normal age-related measures of dopamine (DA) and its metabolites. Adult hm2alpha-SYN-39 transgenic mice had abnormal axons and terminals, age-related impairments in motor coordination, and age-related reductions in DA and its metabolites. Expression of hm2alpha-SYN adversely affects the integrity of dopaminergic terminals and leads to age-related declines in motor coordination and dopaminergic markers.     

GM1 gangliosides alter acute MPTP-induced behavioral and neurochemical toxicity in mice

We investigated the effect of GM1 gangliosides on a 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) animal model of Parkinson disease. Five groups of mice (saline, GM1 (30 mg/kg), MPTP, MPTP + GM1 (15 mg/kg), MPTP + GM1 (30 mg/kg] were compared. GM1 was given daily via intraperitoneal injection before and during 13 daily doses of MPTP (30 mg/kg). Mice were tested for locomotion (1) within 2 h of an MPTP dose (to measure reduced motor activity), and (2) within 24 h of an MPTP dose (after animals had recovered and exhibited hyperactivity). We found that mice given GM1 gangliosides exhibited significantly less MPTP-induced behavior. This effect was most evident with the 15 mg/kg GM1 dose. GM1 also appeared to attenuate MPTP-induced neurochemical changes. GM1 effects indicating enhancement of DA turnover and preservation of DA, DOPAC and HVA concentrations in the striatum were found after the 8th MPTP dose. These latter neurochemical changes, however, were transient and not present after the 13th MPTP dose. Our data would suggest that gangliosides may reduce acute MPTP-induced neurotoxicity in mice either through an increase in DA neuron survival and/or the augmentation of striatal DA activity.     

Behavioral and neurochemical responses in mice bearing tumors submitted to social stress

Through the proinflammatory cytokines secreted in response to inflammation or injury, the immune system produces physiological and behavioral alterations. This study analyzes the effects on behavior, mononuclear proliferative response and central monoamine activity in response to the inoculation of tumor cells in mice submitted to social stress. Two groups of male OF1 mice were used, one of which was inoculated with B16 melanoma cells. Both groups were subdivided into two new groups, with one being submitted to social stress through sensory contact model with a selected aggressive subject, and the other being handled without social interaction. Subjects were exposed to social stress for a 24-h period, with three 5 min intervals of direct physical interaction, where the behavior was recorded and assessed. One hour after the stress and/or handling, they were put down and samples taken for physiological assessment. Significant behavioral changes were found in subjects with implanted tumors, mainly characterized by an increase in avoidance behavior and a decrease in immobility, defense-submission and non-social exploration behavior, coupled with an increase in the spleen mononuclear cell proliferative response. Similarly, an increase was observed in the density of dopamine(2) (D(2))-receptors in the striatum (SRT) and an increase in dopaminergic (DOPAC/DA) and serotonergic (5HIAA/5HT) turnover in the hypothalamus. The increase in the density of D(2)-receptors in the SRT coincides with the decrease in some behaviors with a predominant motor component. The results indicate significant changes in the defensive strategy used to cope with situations of intense social stress in mice bearing tumors.     

Differentiation of neurochemical basis of stress-induced analgesia in mice by selective breeding

Random mated Swiss mice swam for 3 min at 20 °C. The animals with ≤10 s and 50 s postswim latencies on the hot plate (56 °C) were selected as progenitors of low (LA) and high (HA) analgesia lines, respectively. Gradual divergence of latency distributions and postswim hind paw flick latencies between the two lines was observed in successive generations. As shown by the tail flick test applied to the sixth offspring generation, postswim analgesia in the HA line was not only more pronounced but also lasted longer compared to the LA line and the unselected controls. Possible differentiation of the endorphin system activity in the course of selection is discussed.     

Behavioral and neurochemical effects of anpirtoline and citalopram in isolated and group housed mice.

Acute effects of serotonergic drugs acting via different mechanisms were investigated by a social interaction test and subsequent determination of serotonin and dopamine metabolisms in mice housed in groups or isolated for 6 weeks. A resident/intruder test was performed with anpirtoline (5-HT1B receptor agonist in rodents; 1 mg/kg), citalopram (SSRI; 0.5 mg/kg) and saline treatment before animals were decapitated and different brain regions were frozen for subsequent HPLC-analyses. Behavioral investigations indicated a strong increase of aggressive behavior after 6 weeks of isolation housing. Acute citalopram treatment did not influence behavioral parameters of isolated and group housed mice. In contrast, anpirtoline antagonized isolation induced aggressive behavioral components in a specific manner. Analysis of dopamine and serotonin metabolism revealed that citalopram treatment did not affect dopamine metabolism, but reduced serotonin metabolism in the striatum, hippocampus, cortex and midbrain independent of housing conditions. In contrast, anpirtoline treatment increased dopamine metabolism in cortex, striatum and midbrain as well as influenced serotonin metabolism in a structure- and state-specific manner. Whereas anpirtoline decreased serotonin metabolism in the cortex, the midbrain and the hippocampus independent of housing conditions, in the striatum anpirtoline abolished the isolation induced decrease of serotonin metabolism. These results indicate that anpirtoline might induce antiaggressive effects via postsynaptic receptor- and structure-specific activation of serotonergic but also dopaminergic processes, whereas structure independent increase of synaptic serotonin via citalopram was ineffective to reverse aggressivity in isolated mice.     

Effect of brain monoamine precursors on stress-induced behavioral and neurochemical changes in aged mice

Male CF-1 mice aged 22 months showed approximately the same level of motor activity and aggressive behavior as 3-month-old mice under control (no stress) conditions, or 45 min following cold swim stress. Increasing brain catecholamine activity by dietary l-tyrosine treatment had no effect on these two age groups either under control conditions or after stress. In contrast, 30-month-old mice showed lower motor activity under control conditions which was raised significantly by supplementation of the diet with l-tyrosine. However, marked reductions in activity and aggression following stress were observed in the 30-month-old animals and these deficits were not reversed by l-tyrosine treatment prior to stress. Reduction in motor activity was greatest in stressed, 30-month-old mice on l-tyrosine supplemented diets. Compared to 3-month-old mice, the 30-month-old animals had lower brain tyrosine following dietary l-tyrosine treatment, lower brain tryptophan, norepinephrine (NE), dopamine (DA) and DOPAC, but higher HVA, serotonin (5-HT) and 5-HIAA levels. Under both control (no stress) and stress conditions, l-tyrosine pretreatment decreased brain 5-HT in the young animals, but increased 5-HT in the old mice. After stress the 30-month-old animals evidenced only slight increases in levels of blood corticosterone. Brain tyrosine was reduced by stress in the young animals but increased by stress in the old animals. Stress-induced decreases in brain NE and increased in serotonin and 5-HIAA levels were observed in both age groups. These results are consistent with hypotheses concerning age-related alterations in brain monoamine functions and adrenocortical control mechanisms.     

A genetic-correlational study of hippocampal neurochemical variation and variation in exploratory activities of mice.

Previously, we have demonstrated that hippocampal mossy fibers, containing the opioid peptide dynorphin B, are functionally connected with the gene-dependent exploratory behavior of mice. In order to seek further evidence of causal relationships between dynorphin B action and exploration, a quantitative-genetic method was chosen. For this purpose, mice from the inbred strains C57BL/6, DBA/2, BLN, and CPB-K were used. By means of radioimmunoassay, the hippocampal level of dynorphin B was monitored in mice that had been exposed to environmental novelty, as compared to naive animals. Clear evidence was obtained that novelty induces the release of hippocampal dynorphin B. Furthermore, low tissue content was found to be causally connected with high exploratory scores.     

Tiplaxtinin impairs nutritionally induced obesity in mice

To investigate the effect of tiplaxtinin, designed as a synthetic inhibitor of plasminogen activator inhibitor-1 (PAI-1), on obesity, male C57Bl/6 mice (13-14 weeks old) were kept on a high-fat diet (20.1 kJ/g) for four weeks without or with addition of tiplaxtinin (PAI-039) at a dose of 2 mg/g food. At the time of sacrifice, body weights were significantly lower in the inhibitor-treated mice (p < 0.0005). The weights of the isolated subcutaneous and gonadal fat deposits were also significantly lower (both p < 0.0005), associated with adipocyte hypotrophy. Inhibitor-treated adipose tissues displayed similar blood vessel size, but a higher blood vessel density. Fasting glucose and insulin levels, as well as glucose-tolerance tests were not significantly affected by the inhibitor treatment, whereas plasma triglyceride levels were significantly reduced (p = 0.02) and LDL-cholesterol levels significantly enhanced (p = 0.0002). Insulin-tolerance tests revealed significantly lower glucose levels at the end of the test in the inhibitor treated mice (p = 0.03). Thus, in this model of diet-induced obesity in mice administration of tiplaxtinin resulted in impaired adipose tissue development.     

Influence of chronic interleukin-2 infusion and stressors on sickness behaviors and neurochemical change in mice

OBJECTIVES: Major depression is associated with increased circulating interleukin-2 (IL-2) levels, and IL-2 immunotherapy may provoke depressive symptoms, leading to the suggestion that this cytokine may contribute to the evolution of affective disorders. Although depression is a relatively chronic condition, and immunotherapy involves repeated cytokine administration, animal studies have typically assessed the consequences of acute cytokine treatment. The present investigation assessed several behavioral and neurochemical effects of chronic IL-2 infusion. METHODS: Behaviors reflecting anhedonia and/or anorexia, sickness behavior, plasma corticosterone and norepinephrine (NE) activity in the paraventricular nucleus (PVN) of the hypothalamus were assessed following continuous infusion of IL-2 over 7 days in CD-1 mice. RESULTS: The cytokine treatment reduced the consumption of a highly favored palatable substance (chocolate milk) and reduced locomotor activity monitored over the course of the 7-day period. Although sickness behaviors were also increased significantly by the treatment, the degree of sickness behavior was actually modest. While a chronic, variable stressor also affected consumption of the palatable food, this treatment did not enhance the effects of IL-2. Furthermore, in contrast to acute and chronic stressors that increased plasma corticosterone levels and the utilization of NE within the PVN of the hypothalamus, IL-2 did not promote such effects and did not modify the impact of the stressors. CONCLUSION: While IL-2 may induce anorexia or anhedonia, the effects of this treatment are distinguishable from those elicited by stressors and those typically elicited by proinflammatory cytokines. The data are related to findings suggesting a link between IL-2 and depressive illness.     

Behavioral and neurochemical alterations caused by diet restriction - the effect of tyrosine administration in mice

We have investigated the effect of tyrosine administration on the cognitive and neurochemical alterations caused by diet restriction (DR) in mice, as a possible model for some of the behavioral symptoms of patients with anorexia nervosa. Young female mice were fed to 100, 60, and 40% of the calculated daily nutritional requirements for a period of up to 18 days. Cognitive function was evaluated using a modified eight-arm maze with water as a reward. Animals fed to 60% of controls showed significantly improved maze performance while this was impaired in animals on DR to 40%. However, in these animals, injections of tyrosine (100 mg/kg/day) restored performance. Improved maze performance in the 60% DR and 40% DR + tyrosine animals was related to increased beta:alpha tone in the hippocampus - an area, together with the septum, responsible for spatial learning. This was associated with changes in α- and β-receptor density (B_max), without affecting affinity (K_d); and increased norepinephrine (NE) in the 40% DR + tyrosine group, and methoxyhydroxyphenylglycol (MHPG) in both groups. In the hypothalamus, the brain area responsible for energy metabolism, there was a progressive increase in alpha:beta tone with increasing DR associated with changes in B_max. Tyrosine treatment reversed these alterations. Tyrosine improves some of the neurobiological disturbances of DR without causing an increase in body weight. Such a strategy might have important implications for the possible treatment of patients with anorexia nervosa.     

Oxytocin receptor-deficient mice developed late-onset obesity

The oxytocin receptor has been suggested to be involved in energy metabolism, such as food intake and energy consumption. Here, we demonstrate that oxytocin receptor-deficient (Oxtr-/-) male mice exhibited late-onset obesity with increases in abdominal fat pads and fasting plasma triglycerides. Daily food intake and spontaneous motor activity of Oxtr-/- mice were not significantly different as compared with wild-type mice. In contrast, brown adipose tissue in Oxtr-/- mice contained large lipid droplets and cold-induced thermogenesis was impaired. This study demonstrates that oxytocin receptor plays essential roles in the regulation of energy homeostasis.     

Control of hyperphagia prevents obesity in BDNF heterozygous mice

Reduced levels of BDNF in mice cause obesity and behavioral abnormalities including increased aggression and hyperactivity. While it has been shown that the obesity is in part caused by increased food consumption it is still not clear whether defects in other mechanisms involved in the control of body weight homeostasis can also affect this phenotype. Here we report that mice with reduced levels of BDNF do not develop obesity and have normal blood glucose levels if fed over a prolonged period of time the amount of food that control mice usually consume. Thus, hyperphagia appears to be the primary cause of obesity development rather than changes in mechanisms controlling metabolism.     

Altered neurochemical parameters in the brains of mice (Frings) susceptible to audiogenic seizures

In this study, several biochemical aspects of serotonergic neurons were compared in mice susceptible to audiogenic seizures (Frings) and in mice not susceptible to audiogenic seizures (CF 1). Tryptophan hydroxylase (TPH) activity was significantly lower in brains of Frings mice compared to CF1 mice; however, brain levels of 5-HT were similar in both strains. The significantly lower TPH activity in Frings mice was due to the altered kinetic characteristics of TPH activity in this strain. TPH from Frings mice had a significantly lower apparent maximal velocity and a significantly higher affinity for the substrate, as indicated by a lower apparent Km for tryptophan. The uptake of tryptophan and accumulation of 5-HT (following pargyline) were similar in Frings and CF 1 mice. The levels of 5-HT in selected brain regions of Frings and CF 1 mice were also similar, but NE levels were higher in the cerebral cortex and DA levels were higher in the neostriatum of Frings mice. It is unclear at this point what role, if any, 5-HT plays in susceptibility to audiogenic seizures in brains of Frings mice.     

Reduced anxiety‐like behavior and central neurochemical change in germ‐free mice

Background There is increasing interest in the gut‐brain axis and the role intestinal microbiota may play in communication between these two systems. Acquisition of intestinal microbiota in the immediate postnatal period has a defining impact on the development and function of the gastrointestinal, immune, neuroendocrine and metabolic systems. For example, the presence of gut microbiota regulates the set point for hypothalamic‐pituitary‐adrenal (HPA) axis activity. Methods We investigated basal behavior of adult germ‐free (GF), Swiss Webster female mice in the elevated plus maze (EPM) and compared this to conventionally reared specific pathogen free (SPF) mice. Additionally, we measured brain mRNA expression of genes implicated in anxiety and stress‐reactivity. Key Results Germ‐free mice, compared to SPF mice, exhibited basal behavior in the EPM that can be interpreted as anxiolytic. Altered GF behavior was accompanied by a decrease in the N‐methyl‐D‐aspartate receptor subunit NR2B mRNA expression in the central amygdala, increased brain‐derived neurotrophic factor expression and decreased serotonin receptor 1A (5HT1A) expression in the dentate granule layer of the hippocampus. Conclusions & Inferences We conclude that the presence or absence of conventional intestinal microbiota influences the development of behavior, and is accompanied by neurochemical changes in the brain.