Isabel Marian Hartmann Quadros

Brain serotonin receptors and transporters: initiation vs. termination of escalated aggression

RationaleRecent findings have shown a complexly regulated 5-HT system as it is linked to different kinds of aggression.ObjectiveWe focus on (1) phasic and tonic changes of 5-HT and (2) state and trait of aggression, and emphasize the different receptor subtypes, their role in specific brain regions, feed-back regulation and modulation by other amines, acids and peptides.ResultsNew pharmacological tools differentiate the first three 5-HT receptor families and their modulation by GABA, glutamate and CRF. Activation of 5-HT1A, 5-HT1B and 5-HT2A/2C receptors in mesocorticolimbic areas, reduce species-typical and other aggressive behaviors. In contrast, agonists at 5-HT1A and 5-HT1B receptors in the medial prefrontal cortex or septal area can increase aggressive behavior under specific conditions. Activation of serotonin transporters reduce mainly pathological aggression. Genetic analyses of aggressive individuals have identified several molecules that affect the 5-HT system directly (e.g., Tph2, 5-HT1B, 5-HT transporter, Pet1, MAOA) or indirectly (e.g., Neuropeptide Y, αCaMKII, NOS, BDNF). Dysfunction in genes for MAOA escalates pathological aggression in rodents and humans, particularly in interaction with specific experiences.ConclusionsFeedback to autoreceptors of the 5-HT1 family and modulation via heteroreceptors are important in the expression of aggressive behavior. Tonic increase of the 5-HT2 family expression may cause escalated aggression, whereas the phasic increase of 5-HT2 receptors inhibits aggressive behaviors. Polymorphisms in the genes of 5-HT transporters or rate-limiting synthetic and metabolic enzymes of 5-HT modulate aggression, often requiring interaction with the rearing environment.

Can energy drinks reduce the depressor effect of ethanol? An experimental study in mice

Although the popularization of the combined use of alcoholic beverages and energy drinks (ED) containing caffeine, taurine and other substances has increased, there are no controlled experimental studies on the effects of ED alone or combined with ethanol. This work aimed at evaluating the effects of different doses of ED combined or not with ethanol, on the locomotor activity of Swiss mice. The administration of 3.57, 10.71 or 17.86 ml/kg of ED alone increased the locomotor activity of the animals in relation to a control group. Low doses of ethanol (0.5, 1.0 and 1.5 g/kg) alone or in combination with 10.71 ml/kg of ED did not affect their locomotor activity. However, the reduction of activity observed after 2.5 g/kg of ethanol was antagonized by 10.71 ml/kg of ED. Further studies on the mechanisms of this interaction are still needed.

Resistance to ethanol sensitization is associated with increased NMDA receptor binding in specific brain areas.

Co-administration of N-methyl-D-aspartate (NMDA) receptor antagonists is known to block the development of behavioral sensitization to ethanol and other psychostimulants. Since ethanol sensitization in mice does not occur uniformly in all treated animals, the present study examined the possibility that NMDA receptor binding would be selectively altered in mice susceptible to ethanol sensitization. Mice received 2.4 g/kg ethanol or saline i.p. daily for 21 days and were sacrificed 24 h later. No differences in [3H]dizocilpine ([3H](+)MK-801) binding were found between sensitized and vehicle-treated mice in any of the brain regions analyzed. However, ethanol-treated mice that did not develop sensitization showed significantly higher binding in the nucleus accumbens core (+32% and +40% compared to controls and ethanol-sensitized mice, respectively; P<0.04) and the prefrontal cortex (+15% and +22%; P<0.02). In a separate experiment, sensitization resistant mice challenged with 0.25 mg/kg (+)MK 801 showed significantly less motor activation than saline-treated or ethanol-sensitized mice. These results point to a clear association between elevated NMDA receptor binding in specific brain regions and resistance to ethanol sensitization.

Long maternal separation accelerates behavioural sensitization to ethanol in female, but not in male mice

Early life stress is associated with dysfunction of the hypothalamic-pituitary-adrenal (HPA) axis, and with aspects involved in drug abuse. In this study, we investigated the effects of brief (BMS) and long maternal separation (LMS) on the HPA axis response and behavioural sensitization to ethanol (EtOH) in male and female mice. From PND 2 to 14, pups were subjected to daily maternal separation for 15 min (BMS) or 180 min (LMS) or no separated, only handled during cage cleaning (animal facility rearing-AFR). As adults, animals were treated every other day with saline (SAL) or EtOH (2.2g/kg), i.p., for 10 days, and immediately after each administration, their locomotor response was evaluated for 15 min. Forty-eight hours after the 5th administration, all animals were challenged with saline, followed 48 h later, by an EtOH challenge. Corticosterone (CORT) plasma levels were determined 3 times: basal, after the 1st administration and after the EtOH challenge. LMS females showed higher CORT levels than BMS females at basal, but not in response to acute or chronic EtOH administration. The CORT response to EtOH was more robust in LMS and BMS male than AFR male mice. Repeated EtOH treatment induced behavioural sensitization in all groups of male mice. In females, LMS induced a faster sensitization, although BMS females also exhibited behavioural sensitization (4th day and 5th day of treatment, respectively). In conclusion, LMS and BMS produced gender-dependent effects. In females, LMS and BMS facilitated the development of behavioural sensitization, but in the LMS group this effect occurred faster, which may represent increased vulnerability to drug abuse. Moreover, LMS females showed higher basal CORT levels compared to BMS. In males, LMS and BMS increased the CORT response to EtOH but did not modify behavioural sensitization. Therefore, we postulate that LMS female mice exhibited a faster development of behavioural sensitization, but CORT levels were not involved with this effect.

Nucleus accumbens dopamine D 1 receptors regulate the expression of ethanol-induced behavioural sensitization

Repeated ethanol administration may induce behavioural sensitization, defined as a progressive potentiation of locomotor stimulant effects. This process is associated with neuroadaptations in the mesolimbic pathway and the nucleus accumbens. The aim of the present study was to analyse dopamine D₁ receptor (D₁R) participation in locomotor response to an agonist and an antagonist of the D₁R in mice with different levels of sensitization to ethanol. In three separate experiments, mice received administrations of 2.2 g/kg ethanol or saline every other day for 10 d. According to their locomotor response on the last day, ethanol-treated animals were classified into two groups: sensitized or non-sensitized. After the treatment, mice were challenged with 4 or 8 mg/kg SKF-38393 (i.p.), a D₁R agonist (expt 1); or with 0.01 or 0.1 mg/kg SCH-23390 (i.p.), a D₁R antagonist, followed by 2.2 g/kg ethanol (i.p.) administration (expt 2). In expt 3, mice were challenged with intra-accumbens (intra-NAc) SKF-38393 (1 μg/side, in 0.2 μl), and with intra-NAc SCH-23390 (3 μg/side, in 0.2 μl) followed by 2.2 g/kg ethanol (i.p.). Although the i.p. administration of SKF-38393 did not affect the locomotion of mice, the intra-NAc administration of SKF-38393 significantly increased the locomotor activity in sensitized mice, suggesting that sensitized mice present functionally hyperresponsive D₁Rs in the NAc. Both i.p. and intra-NAc administration of SCH-23390 blocked the expression of ethanol sensitization, suggesting that the activation of NAc D₁Rs seems to be essential for the expression of ethanol sensitization.

Individual vulnerability to escalated aggressive behavior by a low dose of alcohol: decreased serotonin receptor mRNA in the prefrontal cortex of male mice

Low to moderate doses of alcohol consumption induce heightened aggressive behavior in some, but not all individuals. Individual vulnerability for this nonadaptive behavior may be determined by an interaction of genetic and environmental factors with the sensitivity of alcohols effects on brain and behavior. We used a previously established protocol for alcohol oral self‐administration and characterized alcohol‐heightened aggressive (AHA) mice as compared with alcohol non‐heightened (ANA) counterparts. A week later, we quantified mRNA steady state levels of several candidate genes in the serotonin [5‐hydroxytryptamine (5‐HT)] system in different brain areas. We report a regionally selective and significant reduction of all 5‐HT receptor subtype transcripts, except for 5‐HT3, in the prefrontal cortex of AHA mice. Comparable gene expression profile was previously observed in aggressive mice induced by social isolation or by an anabolic androgenic steroid. Additional change in the 5‐HT1B receptor transcripts was seen in the amygdala and hypothalamus of AHA mice. In both these areas, 5‐HT1B mRNA was elevated when compared with ANA mice. In the hypothalamus, AHA mice also showed increased transcripts for 5‐HT2A receptor. In the midbrain, 5‐HT synthetic enzyme, 5‐HT transporter and 5‐HT receptors mRNA levels were similar between groups. Our results emphasize a role for postsynaptic over presynaptic 5‐HT receptors in mice which showed escalated aggression after the consumption of a moderate dose of alcohol. This gene expression profile of 5‐HT neurotransmission components in the brain of mice may suggest a vulnerability trait for alcohol‐heightened aggression.

Differential propensity to ethanol sensitization is not associated with altered binding to D1 receptors or dopamine transporters in mouse brain

Behavioral sensitization to ethanols stimulant effect has been proposed as a marker for individual abuse liability. In previous work we have demonstrated that mice showing an increased propensity to EtOH sensitization had higher levels of dopamine (DA) D2 receptor binding in localized brain areas compared to mice showing less sensitization. In the present study we examined whether altered binding to D1 or the DA transporter (DAT) might also be associated with differential propensity to develop EtOH sensitization. Male Swiss mice received 2.4 g/kg EtOH or saline intraperitoneally (i.p.) daily for 21 days, were tested weekly for locomotor activity, and then sacrificed. D1 and DAT binding were assessed by quantitative autoradiography using [3 H]SCH‐23390 and [3 H]WIN 35,428, respectively. EtOH‐treated mice were subdivided into sensitized and non‐sensitized subgroups according to their locomotor activity during treatment. Analyses of brain D1 (19 regions) and DAT (12 regions) binding densities revealed no significant differences among EtOH‐sensitized, ‐non‐sensitized or saline groups in any of the regions measured (all p values > 0.32 for D1 and > 0.16 for DAT). These results suggest that brain D1 and DAT binding, unlike the recently reported changes in D2 binding, do not differentiate mice that develop behavioral sensitization to ethanol from those that do not.

Genetic and Environmental Influences on Ethanol Consumption: Perspectives From Preclinical Research

BACKGROUND Alcohol use disorders (abuse and dependence, AUD) are multifactorial phenomena, depending on the interplay of environmental and genetic variables. METHOD This review describes current developments in animal research that may help (a) develop gene therapies for the treatment of alcoholism, (b) understand the permissive role of stress on ethanol intake, and (c) elucidate why exposure to ethanol early in life is associated with a greater risk of AUD. RESULTS The polymorphisms found in liver alcohol dehydrogenase (ADH) and aldehyde dehydrogenase (ALDH) affect the elimination of ethanol and the susceptibility to ethanol intake. A highly active ADH protects against alcoholism, an effect related to a presteady state burst in arterial acetaldehyde. Social stressors, such as repeated early maternal separation or social defeat, exert a permissive effect on ethanol intake, perhaps by altering the normal development of the hypothalamic-pituitary-adrenal axis. Ethanol exposure during gestation, infancy, or adolescence increases the likelihood of AUD later in life. Early perception of ethanols positive and negative (anti-anxiety) reinforcing effects may play a role in this phenomenon. CONCLUSIONS The review underscores the advantages of using preclinical animal models of AUD and highlights points of intersection between the topics to help design a more integrated approach for the study of alcohol-related problems.

Behavioral and Neurochemical Studies in Distinct Animal Models of Ethanols Motivational Effects

In the last decades, the goal of creating a unique and complete model of alcohol use and alcoholism has been replaced by a myriad of different animal models, each addressing a specific feature of problematic alcohol consumption. This mini-review highlights selected findings in the field of alcohol abuse and dependence, as found through the use of animal models. There are models (e.g., drinking in the dark, drinking after alcohol adulteration or alcohol deprivation) in which animals self-administer alcohol, that are useful to analyze determinants and consequences of binge drinking, progression from casual to problematic alcohol use and relapse or loss of control over alcohol drinking. In other models (e.g., conditioned place preference, conditioned taste aversion, ethanol-induced behavioral sensitization) alcohol dosing is precisely controlled by the experimenter. These models are useful to study motivational (i.e, appetitive, aversive and negative reinforcing) effects of alcohol and neuroadaptive changes that occur after repeated alcohol exposure. The study of age-related differences in reactivity to alcohol provides yet another avenue for analyzing alcohols acute and chronic consequences. Ethanol interacts with several neurotransmitter (dopaminergic, glutamatergic, opioidergic and cannabinoid) and neuromodulators and these interactions are involved in the development and maintenance of alcohol self-administration. The findings described in the review, however, indicate a key role of the endogenous opioid system, notably in the mediation of alcohols postitive rewarding effects. The Review also highlights the need to further assess the inter-relationship between different indices of ethanols motivational effects as well as their association with alcohol intake and preference.

Postsynaptic GABAB Receptor Activity Regulates Excitatory Neuronal Architecture and Spatial Memory

Cognitive dysfunction is a common symptom in many neuropsychiatric disorders and directly correlates with poor patient outcomes. The majority of prolonged inhibitory signaling in the brain is mediated via GABAB receptors (GABABRs), but the molecular function of these receptors in cognition is ill defined. To explore the significance of GABABRs in neuronal activity and cognition, we created mice with enhanced postsynaptic GABABR signaling by mutating the serine 783 in receptor R2 subunit (S783A), which decreased GABABR degradation. Enhanced GABABR activity reduced the expression of immediate-early gene-encoded protein Arc/Arg3.1, effectors that are critical for long-lasting memory. Intriguingly, S783A mice exhibited increased numbers of excitatory synapses and surface AMPA receptors, effects that are consistent with decreased Arc/Arg3.1 expression. These deficits in Arc/Arg3.1 and neuronal morphology lead to a deficit in spatial memory consolidation. Collectively our results suggest a novel and unappreciated role for GABABR activity in determining excitatory neuronal architecture and spatial memory via their ability to regulate Arc/Arg3.1.