Drug-induced Behavioral Changes Research Articles

Evaluation and comparison of pharmacokinetic profiles and safety of two extended-release buprenorphine formulations in common marmosets (Callithrix jacchus)

While sustained-release buprenorphine (BSR) is used as a long-lasting opioid analgesic in common marmosets (Callithrix jacchus), there are no published studies on pharmaceutical-grade extended-release buprenorphine options such as Ethiqa XR (EXR) for this species. However, BSR is a compounded product and has been reported to cause injection site reactions in multiple species, including marmosets. Additionally, now with the availability of EXR, a pharmaceutical-grade veterinary product, the use of BSR in laboratory animals is not compliant with the Guide for the Care and Use of Laboratory Animals (Guide) unless scientifically justified and approved by the IACUC. We compared pharmacokinetic and safety profiles of BSR (0.15 mg/kg) and EXR (0.1–0.2 mg/kg) administered subcutaneously to adult marmosets. Blood was collected by venipuncture of the saphenous vein at multiple time points (0.25–72 h) and analyzed by liquid chromatography-tandem mass spectrometry (LC–MS/MS). EXR between 0.1 and 0.2 mg/kg resulted in a dose-dependent increase in Cmax (1.43–2.51 ng/mL) and were not statistically different from BSR (1.82 ng/mL). Tmax, lambdaz, and t1/2 were not statistically different between formulations. Mean plasma buprenorphine concentrations for BSR and EXR exceeded the therapeutic threshold (0.1 ng/mL) within 0.25 h and lasted for > 72 h. Mild sedation, but neither respiratory depression nor ataxia, was observed for both formulations. BSR injection sites had significantly higher histopathological scores compared to EXR. Video recordings for monitoring drug-induced behavioral changes showed increased animal activity levels after BSR and EXR versus saline controls. Norbuprenorphine, a buprenorphine metabolite associated with respiratory depression, was detected in the plasma after BSR and EXR administration as well as by in vitro liver microsome assays. In conclusion, we recommend using EXR over BSR as a long-lasting buprenorphine analgesic in marmosets because EXR is a pharmaceutical-grade formulation that is compliant with FDA guidelines and the Guide as well as exhibits comparable PK and safety profiles as BSR.

Knockout of Dopamine D3 Receptor Gene Blocked Methamphetamine-Induced Distinct Changes of Dopaminergic and Glutamatergic Synapse in the Nucleus Accumbens Shell of Mice.

Structural plasticity changes in the brain are thought to underlie, at least partially, drug-induced persistent changes in behavior. Our previous study reported that increased synaptic density in the nucleus accumbens shell (NAcsh) correlates with and may contribute to behavioral sensitization induced by methamphetamine (METH). However, the distinct changes of dopaminergic and glutamatergic synapses and the modulating effects of dopamine D3 receptor remain unclear. In the current study, we used immunohistochemistry electron-microscopy and immunofluorescence to detect the changes of dopamine D1, D2, and glutamate NR2B-positive synapses and cells in the NAcsh of METH-sensitized wild type (WT) and knockout of dopamine D3 receptor gene (D3–/–) mice. We found that METH induced long-term behavioral sensitization in WT mice, which was accompanied by an increased number and rate of dopamine D1 receptor-positive synapses and cells, as well as glutamate NR2B-positive synapses and cells. In contrast, the number and rate of dopamine D2 receptor-positive synapses and cells were significantly decreased in the NAcsh of METH-sensitized WT mice. D3–/– mice exhibited attenuated acute locomotor responses and behavioral sensitization to METH compared with WT mice. Moreover, the knockout of dopamine D3 receptor gene inhibited METH-induced changes of dopaminergic and glutamatergic synapses in the NAcsh of METH-sensitized mice. Taken together, our results suggest that METH induced distinct changes of dopaminergic and glutamatergic synapses and cells in the NAcsh of mice, which was blocked by the knockout of dopamine D3 receptor gene, and may contribute to, at least partially, METH-induced behavior sensitization as well as the modulating effect of the dopamine D3 receptor.

The cognitive and behavioral effects of D-amphetamine and nicotine sensitization in adult zebrafish

BackgroundZebrafish are growing in use as a model for understanding drug dependence and addiction. Sensitization paradigms have been a useful tool in identifying mechanisms involved in drug-induced behavioral and neurological changes, but in zebrafish have tended to focus on locomotor, rather than cognitive, endpoints.MethodsHere, we used a novel method, the FMP Y-maze, which measures continuous performance through a series of repeated binary choices (L vs R), to establish a model for assessing parameters associated with psychostimulant-induced behavioral and cognitive sensitization in adult zebrafish.ResultsRepeat, intermittent exposure to d-amphetamine (AMPH) for 14 days increased alternations (LRLR) in the maze, suggesting improved working memory, which was enhanced further following drug challenge after a short withdrawal period, suggesting behavioral sensitization. However, this cognitive enhancement coincided with a reduction in the use of other exploration strategies, hypolocomotion, and inhibition of cognitive flexibility. Like AMPH, exposure to nicotine (NIC) increased alternations following drug challenge after chronic treatment. Repeat NIC exposure appeared to induce both cognitive and psychomotor sensitization, as evidenced by increased working memory performance (alternations) and locomotor activity, without negatively impacting other search strategies or cognitive flexibility.ConclusionChronic treatment with AMPH or NIC boosts cognitive performance in adult zebrafish. Cognitive sensitization occurred with both drugs, resulting in enhanced working memory; however, repeat AMPH exposure, following a withdrawal period, resulted in inhibited cognitive flexibility, an effect not evident with repeat NIC exposure. Cognitive and behavioral sensitization paradigms in zebrafish could serve as a useful tool for assessing cognitive states which result in cognitive enhancing or impairing effects of drugs.

A novel rodent accelerometer device for measuring locomotor activity and other movements

Locomotor activity is a simple, yet useful measure of drug-induced changes in rodent behavior. Current methods for assessing activity often fail to capture more complex behavioral topographies, such as stereotypy, require measuring movement in novel environments, and/or require invasive surgeries. Here, we describe a novel device that wirelessly transmits fine motor behaviors at high frequencies. An accelerometer (Biosense, Palo Alto, CA) encased in a plastic covering (2 cm × 1 cm × 0.5 cm) is adhered to the skin between the scapula of a rodent. The device records changes in acceleration (m/s2) and converts it to frequency (Hz) using a Fourier transform. Data is collected about 50,000 times per 10 minutes and is sent to a computer via WiFi. Once new data is uploaded to the cloud (Google Cloud), a system processes the raw accelerometer values and quantifies certain types of activity using proprietary Python code. The processed data is accessible via an online web portal. To compare this novel approach with established methods, male Sprague Dawley rats were habituated to a novel acrylic cage (14” × 14” × 8”) for 90 minutes then were treated with amphetamine (0, 0.32, 1.0, 3.2 mg/kg subcutaneous, n=8 per dose). Locomotor activity was recorded using infrared beam breaks (Columbus Instruments, Columbus, OH) and the accelerometers simultaneously in the same rats. Amphetamine dose-dependently increased locomotor activity in both measures for approximately 2.5 hours, with the magnitude and duration of the effect being closely correlated between the two measures. Further, only the accelerometers were able to identify and quantify stereotyped movements and other behaviors, such as walking, running, or wet-dog shakes. In addition to locomotor activity, the devices were able to detect pentylenetetrazole-induced convulsion related movements (56 mg/kg subcutaneous, n=6) and increased activity during the dark cycle as compared with light cycle in the home cage (n=8). This novel method for recording rodent activity will be a useful approach for capturing and quantifying complex drug-induced behaviors as well as changes in circadian behavior in known and novel environments.

Imaging Fast-Acting Drug Effects in Humans Using 1H-MRS.

Proton magnetic spectroscopy (1H-MRS) is a noninvasive imaging technique that allows for the quantification of neurometabolic compounds at millimolar concentrations in the living human brain. This technique has been most often used to assess long-term changes in human brain metabolism in psychiatric disorders, pharmacological treatment, chronic drug use, and alcohol dependence. In contrast, the capacity of 1H-MRS to evaluate the biochemical changes in the minutes to hours following drug consumption, which contribute to fast-acting drug-induced changes in perception, mood, cognition, and behavior, is largely unexplored. This Viewpoint highlights the utility of 1H-MRS imaging for revealing neural mechanisms of rapid drug action in the human brain, with implications for phasic, in vivo changes in biosynthetic and catabolic pathways after drug exposure. Drawing from examples of psychostimulant drug effects, neuromodulatory input and drug-induced mood, we present strategies to optimize 1H-MRS for noninvasively imaging fast-acting drug effects and other rapid phenomena within the living human brain. These approaches could provide powerful tools for both basic research and drug development.

Use of the Actiwatch-Mini® in dog safety studies as an early indicator for drug-induced behavioural changes

IntroductionClinical observations are routinely used to address drug-induced behavioural changes in dogs. To deal with the limitations of this methodology, we evaluated Actiwatch-Mini® as a tool to monitor continuously locomotor activity in dogs and to provide objective parameters that could be linked to behavioural changes after compound administration. MethodsEight Beagles were equipped with Actiwatch-Mini®. As a reference drug, a PDE2-inhibitor was administered daily for six days at doses known to cause minor or severe behavioural changes. ResultsWhile traditional observation showed no behavioural changes - except sedation - dogs receiving 0.5 mg/kg/day, showed significant increases in % immobile time (+15.8%) and mean length of immobile phases (+1.2 min) but no change in number of immobile phases (+2.2). At 1 mg/kg/day, light to severe changes in behaviour were present. Actiwatch-Mini® recorded an increase in mean length of immobile phases (+1.9 min) and a decrease in mean number of immobile phases (−7.4) in the first four hours after each dosing while total % immobile time was not significantly increased (+4.9%). DiscussionActiwatch-Mini® was able to detect changes in immobility parameters in dogs dosed with a PDE2-inhibitor while no or only minor behavioural changes were observed. The system can be used for continuously monitoring the activity of dogs, to provide objective scores for evaluation of activity. It provides an inexpensive and low labour-intensive method for detecting changes in activity and possible early indications of drug-induced behavioural changes.

Analysis of behavioral changes in zebrafish (Danio rerio) larvae caused by aminoglycoside-induced damage to the lateral line and muscles

Zebrafish behavior is influenced by the lateral line hair cells and muscles. Drug-induced behavioral changes can serve as indicators in the evaluation of drug toxicity. The aminoglycoside family of antibiotics comprise a number of agents, including neomycin (NM) and gentamicin (GM). We hypothesized that NM and GM exert different effects on zebrafish larvae through their action on the lateral line and muscle fibers, inducing different swimming behavioral patterns such as locomotor behavior and the startle response. In this study, 125 μM NM and 5, 10, 20 μM GM induced hair cell damage in the anterior and posterior lateral lines of zebrafish larvae. However, unlike GM, 125 μM NM also caused muscle damage. Locomotor behavior was decreased in the 125 μM NM-exposed group compared to the group exposed to GM. Furthermore, 125 μM NM exposure induced significantly different patterns of various indices of startle behavior compared with the GM exposure groups. Additionally, the larvae exhibited different startle responses depending on the concentration of GM. These results suggest that GM may be the drug-of-choice for analyzing behavioral changes in zebrafish caused by damage to the lateral line alone. Our study highlights the importance of confirming muscle damage in behavioral analyses using zebrafish.

Quantifying free behaviour in an open field using k-motif approach

Quantification and parametrisation of movement are widely used in animal behavioural paradigms. In particular, free movement in controlled conditions (e.g., open field paradigm) is used as a “proxy for indices of baseline and drug-induced behavioural changes. However, the analysis of this is often time- and labour-intensive and existing algorithms do not always classify the behaviour correctly. Here, we propose a new approach to quantify behaviour in an unconstrained environment: searching for frequent patterns (k-motifs) in the time series representing the position of the subject over time. Validation of this method was performed using subchronic quinpirole-induced changes in open field experiment behaviours in rodents. Analysis of this data was performed using k-motifs as features to better classify subjects into experimental groups on the basis of behaviour in the open field. Our classifier using k-motifs gives as high as 94% accuracy in classifying repetitive behaviour versus controls which is a substantial improvement compared to currently available methods including using standard feature definitions (depending on the choice of feature set and classification strategy, accuracy up to 88%). Furthermore, visualisation of the movement/time patterns is highly predictive of these behaviours. By using machine learning, this can be applied to behavioural analysis across experimental paradigms.

Drug-induced changes in mechanical behavior of electrically synchronized cardiomyocytes on surface-patterned polydimethylsiloxane diaphragm

We herein describe the fabrication and evaluation of an electrical stimulator-integrated functional polydimethylsiloxane (PDMS) diaphragm platform to examine the influence of drug toxicity on electrically synchronized cardiomyocytes. The integrated electrical stimulator allows the cardiomyocytes to contract and relax at a specific beat frequency. The growth conditions of the cardiomyocytes can be modulated by controlling the duty cycle, pulse duration, and frequency of the electrical stimulator. Electrical stimulation not only synchronized the beat cycle of the cardiomyocytes but also affected cardiomyocyte maturation. The microgroove structures formed on the PDMS surface had a positive influence on the contractile force through the alignment of the cardiomyocytes. The displacement of the PDMS diaphragm caused by the contractile characteristics of the cardiomyocytes was precisely measured at the nanoscale using a laser displacement sensor. After preliminary experiments were carried out using the platform, two typical cardiovascular drugs (Verapamil and E-4031) were selected to evaluate the performance of the proposed sensor platform. The changes in contraction and beat rate due to drug toxicity were systematically investigated in the electrically synchronized and spontaneously beating cardiomyocytes on the PDMS diaphragm. The use of electrically synchronized cardiomyocytes in drug toxicity screening may contribute to elucidating the relationship between cellular contraction and drugs.

Combined effect of gestational stress and postpartum stress on maternal care in rats

Variations in maternal care in the rat influence the development of individual differences in behavioral and endocrine responses to stress. This study aimed to examine the interaction between intragastric intubation during late gestation and postpartum stress, induced by pup separation, on maternal behavior and on dams' emotional state and HPA axis function. Rats received intragastric intubation of water on days 12–20 of gestation or remained untreated in their home cage (naïve dams). Pup separation was used as a model of postpartum stress. The procedure consisted of a daily separation of the dam from its litter for 3h from PND 3 until PND 15. Pup separation was carried out in both naïve and intubated dams. The behavioral results indicate that the association of these two stressors significantly decreased arched-back nursing (ABN) and licking and grooming (LG), behaviors considered important parameters to discriminate the high quality of maternal care. Moreover, dams that received both stressors displayed less nest building and blanket nursing behaviors; no effect on the frequency of passive and total nursing was recorded. The analysis of single effects on ABN and LG, revealed that dams that underwent gestational stress induced by intragastric intubation displayed less LG, but ABN was overall unchanged. On the contrary, pup separation stress significantly increased ABN and LG upon reunion of naïve dams with their pups. Treatments per se or the association of both induced modest changes in plasma levels of allopregnanolone and corticosterone that likely did not influence maternal care. These data show that the association of a mild stress during gestation with an unfavorable experience after parturition had a significant impact on maternal care. This effect seems independent from HPA axis activation or from changes in emotional state; further studies would be necessary to ascertain the neural changes that could contribute to altered maternal behavior in stressed mothers. Moreover, these results suggest that the use of intragastric intubation during gestation would interfere with measures of drug-induced changes in maternal behavior and likely their consequences on the offspring.

Alterations in Striatal Circuits Underlying Addiction-Like Behaviors.

Drug addiction is a severe psychiatric disorder characterized by the compulsive pursuit of drugs of abuse despite potential adverse consequences. Although several decades of studies have revealed that psychostimulant use can result in extensive alterations of neural circuits and physiology, no effective therapeutic strategies or medicines for drug addiction currently exist. Changes in neuronal connectivity and regulation occurring after repeated drug exposure contribute to addiction-like behaviors in animal models. Among the involved brain areas, including those of the reward system, the striatum is the major area of convergence for glutamate, GABA, and dopamine transmission, and this brain region potentially determines stereotyped behaviors. Although the physiological consequences of striatal neurons after drug exposure have been relatively well documented, it remains to be clarified how changes in striatal connectivity underlie and modulate the expression of addiction-like behaviors. Understanding how striatal circuits contribute to addiction-like behaviors may lead to the development of strategies that successfully attenuate drug-induced behavioral changes. In this review, we summarize the results of recent studies that have examined striatal circuitry and pathway-specific alterations leading to addiction-like behaviors to provide an updated framework for future investigations.

Drug-Induced Behavioral Changes: Using Laboratory Observations to Predict Field Observations

Behavioral assays constitute important research tools when assessing how fish respond to environmental change. However, it is unclear how behavioral modifications recorded in laboratory assays are expressed in natural ecosystems, a limitation that makes it difficult to evaluate the predictive power of laboratory-based measurements. In this study, we hypothesized that exposure to a benzodiazepine (i.e., oxazepam) increases boldness and activity in laboratory assays as well as in field assays – that is, laboratory results can be used to predict field results. Moreover, we expected the modified behavior to affect other important ecological measures such as habitat selection and home range. To test our hypothesis, we exposed European perch (Perca fluviatilis) to oxazepam and measured subsequent changes in behavioral trials both in laboratory assays and in a lake ecosystem populated with a predatory fish species, pike (Esox lucius). In the lake, the positions of both perch and pike were tracked every three minutes for a month using acoustic telemetry. In the laboratory assay, the oxazepam-exposed perch were bolder and more active than the non-exposed perch. In the lake assay, the oxazepam-exposed perch were also more bold and active, had a larger home range, and used pelagic habitats more than the non-exposed perch. We conclude that ecotoxicological behavioral assays are useful for predicting the effects of exposure in natural systems. However, although individual responses to exposure were similar in both the laboratory and field trials, effects were more obvious in the field study, mainly due to reduced variability in the behavior measures from the lake. Hence, short-term behavioral assays may fail to detect all the effects expressed in natural environments. Nevertheless, our study clearly demonstrates that behavior modifications observed in laboratory settings can be used to predict how fish perform in aquatic ecosystems.

Morphine-induced inhibition of Ca2+ -dependent d-serine release from astrocytes suppresses excitability of GABAergic neurons in the nucleus accumbens.

The nucleus accumbens (NAc) plays a critical role in addictive drug-induced behavioral changes. d-serine is present at high levels in the brain and is involved in the regulation of N-methyl-d-aspartate glutamate (NMDA)-dependent synaptic activity. In this study, we aimed to examine the involvement of d-serine in morphine addiction. Morphine decreased the NMDA receptor-mediated excitatory postsynaptic currents and excitability of GABAergic neurons in the NAc, while exogenous d-serine alleviated the effects of morphine. Morphine reduced extracellular d-serine levels in rat NAc or in primary culture of astrocytes through inhibition of intracellular Ca2+ signals and blockade of d-serine release from cell vesicles. Morphine induced robust internalization of α-amino-3-hydroxy-5-methyl-4-isoxazole-propionate acid receptor (AMPAR) in primary cultured astrocytes. Moreover, administration of exogenous d-serine to rats inhibited the development of locomotor sensitization to morphine, attenuated the morphine-induced potentiation on conditioned place preference and suppressed the morphine-enhanced expression of p-CREB and ΔFosB in the NAc. Overall, our results showed that morphine inhibited d-serine release from astrocytes through modulation of AMPAR-mediated Ca2+ influx, and led to the inhibition of postsynaptic excitability of GABAergic neurons in the NAc. This work may provide a new insight into the underlying mechanisms of morphine addiction.

`Up-regulation of histone acetylation induced by social defeat mediates the conditioned rewarding effects of cocaine

Social defeat (SD) induces a long-lasting increase in the rewarding effects of psychostimulants measured using the self-administration and conditioned place procedures (CPP). However, little is known about the epigenetic changes induced by social stress and about their role in the increased response to the rewarding effects of psychostimulants. Considering that histone acetylation regulates transcriptional activity and contributes to drug-induced behavioral changes, we addressed the hypothesis that SD induces transcriptional changes by histone modifications associated with the acquisition of place conditioning. After a fourth defeat, H3(K9) acetylation was decreased in the hippocampus, while there was an increase of HAT and a decrease of HDAC levels in the cortex. Three weeks after the last defeat, mice displayed an increase in histone H4(K12) acetylation and an upregulation of histone acetyl transferase (HAT) activity in the hippocampus. In addition, H3(K4)me3, which is closely associated with transcriptional initiation, was also augmented in the hippocampus three weeks after the last defeat. Inhibition of HAT by curcumin (100mg/kg) before each SD blocked the increase in the conditioned reinforcing effects of 1mg/kg of cocaine, while inhibition of HDAC by valproic acid (500mg/kg) before social stress potentiated cocaine-induced CPP. Preference was reinstated when animals received a priming dose of 0.5mg/kg of cocaine, an effect that was absent in untreated defeated mice. These results suggest that the experience of SD induces chromatin remodeling, alters histone acetylation and methylation, and modifies the effects of cocaine on place conditioning. They also point to epigenetic mechanisms as potential avenues leading to new treatments for the long-term effects of social stress on drug addiction.

Differential activation of dopaminergic systems in rat brain basal ganglia by morphine and methamphetamine

Typical abused drug-induced behavioral changes are ordinarily mediated by the mesolimbic dopaminergic system and even the phenotypes of behavior are different from each other. However, the mechanisms that underlie the behavioral changes induced by these abused drugs have not yet been elucidated. The present study was designed to investigate the mechanisms that underlie how abused drugs induce distinct behavioral changes using neurochemical as well as behavioral techniques in rats. Methamphetamine (2mg/kg) more potently increased dopamine release from the striatum more than that from the nucleus accumbens. In contrast, the administration of morphine (10mg/kg) produced a significant increase in the release of dopamine from the nucleus accumbens, but not the striatum, which is accompanied by a decrease in the release of GABA in the ventral tegmental area. These findings indicate that morphine and methamphetamine differentially regulate dopaminergic systems to produce behavioral changes, even though both drugs have abuse potential through activation of the mesolimbic dopaminergic system.

Epigenetics and drug addiction: a focus on MeCP2 and on histone acetylation

Chronic drug exposure alters gene expression in the brain, which is believed to underlie compulsive drug seeking and drug taking behavior. Recent evidence shows that drug-induced long-term neuroadaptations in the brain are mediated in part by epigenetic mechanisms. By remodeling chromatin, this type of regulation contributes to drug-induced synaptic plasticity that translates into behavioral modifications. How drug-induced alterations in DNA methylation regulate gene expression is reviewed here, with a focus on MeCP2, a protein binding methylated DNA. The importance of histone modifications, especially acetylation is also discussed, with an emphasis on the effects of inhibitors of histone deacetylases on drug-induced behavioral changes. The precise identification of the epigenetic mechanisms that are under the control of drugs of abuse may help to uncover novel targets for the treatment of drug seeking and relapse.

Effects of amphetamine exposure in adolescence or young adulthood on inhibitory control in adult male and female rats

Heightened impulsivity is a feature of some psychiatric disorders, including addiction, that also have sex-specific patterns of expression. The relationship between addiction and impulsivity may be driven by drug-induced changes in behavior caused by long term adaptations in signaling within the medial prefrontal cortex (mPFC). Here, we used a response inhibition task that is sensitive to changes in mPFC function to examine the effects of sex and exposure to amphetamine (AMPH) on impulsive action and vigilance. We also examined drug-induced alterations in glutamatergic and dopaminergic signaling through challenge injections with the NMDA receptor antagonist MK-801 (dizocilpine) and AMPH. Male and female Sprague Dawley rats were injected (i.p.) with saline or 3mg/kg AMPH every other day during adolescence (postnatal day (P) 27–45) or adulthood (P85–103). Starting on P125–135, rats were tested for their ability to lever press for a food reward during periods of signaled availability and withhold responding during a “premature response” phase. In experiment 1, rats received challenge injections (i.p.) of MK-801 and AMPH followed by tests of task performance and locomotor activity. In experiment 2, rats received intra-mPFC infusion of MK-801. We found that females had better inhibitory control and poorer vigilance than males and that AMPH exposure had both sex- and age-of-exposure dependent effects on impulsivity. Systemic drug challenges disrupted task performance, particularly in females, and increased impulsivity while intra-mPFC infusions had modest effects. AMPH exposure did not affect responses to drug challenges. Together, these results suggest that sex mediates both trait and drug-induced impulsivity.

Combinational effects of ketamine and amphetamine on behaviors and neurotransmitter systems of mice

The combined ingestion of ketamine (Ket) and amphetamine (Amph) by drug-users has been rampant and produced more severe behavioral abnormality. However, the interactive consequences of the two drugs are still unclear. In this study, we treated adult male mice with a single i.p. injection of saline, Amph (5mg/kg), low Ket (LK, 10mg/kg), high Ket (HK, 50mg/kg), or Amph and LK or HK (ALK or AHK) and examined their behavioral and neurochemical changes at 0.5 and 2h post-injection. Compared with saline, Amph, LK or HK treatment alone increased the levels of motor activities such as locomotion, stereotypy or ataxia of mice. Notably, at combined treatments, LK and HK differentially exacerbated Amph-induced locomotion and stereotypy, whereas Amph worsened LK or HK-produced ataxia. The higher striatal dopamine levels of A, ALK and AHK groups correlated with their greater motor activities. The prolonged increase of dopamine in the motor cortex of ALK and AHK mice may associate with the longer duration of behavioral hyperactivity and greater peak score of locomotion; the greater dopamine level in the somatosensory cortex probably contributes to the more severe ataxia. Furthermore, in the striatum of all drug-treated groups, the expression of GAD67 mRNA and GAD67-positive punctates was higher than respective saline controls, indicating the involvement of GABAergic system in the drug-induced behavioral changes. Our results demonstrate the acute interplay between Amph and Ket in both behavioral and neurochemical aspects for the first time. Dopaminergic and GABAergic systems were affected differentially by the drugs in the striatum.

Effects of morphine on immediate-early gene expression in the striatum of C57BL/6J and DBA/2J mice

BackgroundImmediate early gene (IEG) induction elicited by drugs of abuse may contribute to development of plastic changes in the brain responsible for drug-induced behavioral changes leading to addiction. The aim of the present study was to characterize the changes in IEG expression in the striatum and nucleus accumbens produced by an acute or chronic administration of morphine. MethodsIn order to search for a possible relationship between morphine-induced IEG expression and behavior, the experiment was performed on two inbred strains of mice, C57BL/6J and DBA/2J, which differ markedly in their sensitivity to the rewarding and locomotor stimulatory actions of opiates. Gene expression was assessed using RT-PCR and DNA microarrays. ResultsThe experiments demonstrated a prolonged or a delayed up-regulation of 14 IEG in the striatum at 4h after morphine administration. Among them, a cluster of 8 genes, including 6 inducible transcription factors (c-fos, fra-2, junB, zif268 (egr1), egr2, NGFI-B) and 2 effector IEG (arc and mkp1) seemed to be regulated in concert in response to morphine. This group of genes was induced to a greater degree after chronic than acute morphine administration selectively in C57BL/6J mice and the difference bore apparently no relationship to opiate-produced locomotor activation. The strain-selective regulation was also demonstrated for cyclin L2 and tPA after an acute morphine injection. ConclusionsOur data indicate that morphine up-regulates many IEG in the mouse striatum at a strikingly delayed time-point and that these changes are genotype-dependent. They also suggest inter-strain differences in the development of striatal neuroadaptations to chronic morphine treatment.

Effect of Risperidone and Fluoxetine on the Movement and Neurochemical Changes of Zebrafish

Brain developmental disorders in humans, including Autism Spectrum Disorders (ASD) and Down’s syndrome, have been linked to increased serotonin levels. This work was designed to study changes in serotonin levels in the early stages of development with two classes of antipsychotic drugs: Risperidone, a drug that blocks serotonin and dopamine receptors, and fluoxetine, a serotonin reuptake inhibitor. The use of antipsychotic drugs is a solid choice to study the decrease and increase of these neurotransmitters and their influence on development. The study of these parameters will give an idea of the effects of serotonin in early developmental stages. To this end, we examined the effects of risperidone and fluoxetine on the locomotor activity, heart rate and brain development of zebrafish larvae. Our results showed that in larvae exposed to fluoxetine alone, swimming was significantly increased at 9 dpf (days post-fertilization). Erratic and abnormal movements were observed suggesting a toxic effect of fluoxetine. No erratic swimming was observed in larvae treated with fluoxetine plus risperidone. Both drugs presented morphological changes in dopaminergic neurons and mononeurons. Exposure to fluoxetine plus risperidone indicated possible reversal effects. Studies in zebrafish allow obtaining new insights into the side effects of these drugs as well as into the brain control of locomotor activity. Testing several drug-induced changes in behavior and serotonin levels is one of the experimental approaches for screening a new therapeutically relevant compound, and thus, merits further research.