Negative impacts of social isolation on behavior and neuronal functions are recovered after short-term social reintroduction in zebrafish.

Recently, social isolation measures were crucial to prevent the spread of the coronavirus pandemic. However, the lack of social interactions affected the population mental health and may have long-term consequences on behavior and brain functions. Here, we evaluated the behavioral, physiological, and molecular effects of a social isolation (SI) in adult zebrafish, and whether the animals recover such changes after their reintroduction to the social environment. Fish were submitted to 12 days of SI, and then reintroduced to social context (SR). Behavioral analyses to evaluate locomotion, anxiety-like and social-related behaviors were performed after SI protocol, and 3 and 6 days after SR. Cortisol and transcript levels from genes involved in neuronal homeostasis (c-fos, egr, bdnf), and serotonergic (5-HT) and dopaminergic (DA) neurotransmission (thp, th) were also measured. SI altered social behaviors in zebrafish such as aggression, social preference, and shoaling. Fish submitted to SI also presented changes in the transcript levels of genes related to neural activity, and 5-HT/DA signaling. Interestingly, most of the behavioral and molecular changes induced by SI were not found again 6 days after SR. Thus, we highlight that SR of zebrafish to their conspecifics played a positive role in social behaviors and in the expression of genes involved in different neuronal signaling pathways that were altered after 12 days of SI. This study brings unprecedented data on the effects of SR in the recovery from SI neurobehavioral alterations, and reinforces the role of zebrafish as a translational model for understanding the neurobiological mechanisms adjacent to SI and resocialization.

Zebrafish as a potential non-traditional model organism in translational bipolar disorder research: Genetic and behavioral insights.

Bipolar disorder (BD) is a severe and debilitating illness that affects 1-2% of the population worldwide. BD is characterized by recurrent and extreme mood swings, including mania/hypomania and depression. Animal experimental models have been used to elucidate the mechanisms underlying BD and different strategies have been proposed to assess BD-like symptoms. The zebrafish (Danio rerio) has been considered a suitable vertebrate system for modeling BD-like responses, due to the genetic tractability, molecular/physiological conservation, and well-characterized behavioral responses. In this review, we discuss how zebrafish-based models can be successfully used to understand molecular, biochemical, and behavioral alterations paralleling those found in BD. We also outline some advantages and limitations of this aquatic species to examine BD-like phenotypes in translational neurobehavioral research. Overall, we reinforce the use of zebrafish as a promising tool to investigate the neural basis associated with BD-like behaviors, which may foster the discovery of novel pharmacological therapies.

Ketamine acutely impairs memory consolidation and repeated exposure promotes stereotyped behavior without changing anxiety- and aggression-like parameters in adult zebrafish.

Ketamine is a dissociative anesthetic in human and veterinary clinic, as well as an abuse drug that acts on several neurotransmitter systems. The use of alternative animal models, such as zebrafish, is emerging to study the effects of drugs on neurobehavioral responses. Here, we evaluated the effects of ketamine on memory consolidation (acute protocol), as well as on anxiety-, aggressive-like behavior, and whole-body cortisol levels in adult zebrafish after a repeated exposure. For the acute protocol, fish were tested in the inhibitory avoidance task (training and testing with a 24-hour interval). Immediately after the training session, fish were exposed to ketamine (0, 2, 20, or 40 mg/L) for 20 min. The exploratory activity was also measured 24 h after acute exposure to exclude the influence of impaired locomotion on memory performance. For the repeated exposure, animals were exposed to the same concentrations of ketamine for 20 min (7 days). After the last exposure (24 h later), anxiety- and aggression-like behaviors were quantified in the novel tank and mirror-induced aggression tests, respectively, as well as whole-body cortisol levels measurements were performed. The highest ketamine concentration tested (40 mg/L) acutely induced a slight memory impairment in the inhibitory avoidance task without changing locomotion and anxiety-like behaviors. Although locomotion, anxiety-, aggressive-like behaviors, and whole-body cortisol levels did not change after repeated exposure, 40 mg/L ketamine increased circling behavior. Overall, our data reinforce that ketamine acutely affects multiple behavioral domains in zebrafish, in which repeated ketamine exposure elicits stereotyped behavior, without changing locomotion, aggression, and anxiety/stress-related parameters.

Using zebrafish (Danio rerio) models to understand the critical role of social interactions in mental health and wellbeing.

Social behavior represents a beneficial interaction between conspecifics that is critical for maintaining health and wellbeing. Dysfunctional or poor social interaction are associated with increased risk of physical (e.g., vascular) and psychiatric disorders (e.g., anxiety, depression, and substance abuse). Although the impact of negative and positive social interactions is well-studied, their underlying mechanisms remain poorly understood. Zebrafish have well-characterized social behavior phenotypes, high genetic homology with humans, relative experimental simplicity and the potential for high-throughput screens. Here, we discuss the use of zebrafish as a candidate model organism for studying the fundamental mechanisms underlying social interactions, as well as potential impacts of social isolation on human health and wellbeing. Overall, the growing utility of zebrafish models may improve our understanding of how the presence and absence of social interactions can differentially modulate various molecular and physiological biomarkers, as well as a wide range of other behaviors.

Taurine modulates behavioral effects of intermittent ethanol exposure without changing brain monoamine oxidase activity in zebrafish: Attenuation of shoal- and anxiety-like responses, and abolishment of memory acquisition deficit.

Prolonged alcohol consumption has been considered as an important risk factor for various diseases. Chronic ethanol (EtOH) intake is associated with deleterious effects on brain functions culminating in robust behavioral changes. Notably, drugs available to treat the effects of EtOH have low therapeutic efficacy so far. Taurine (TAU) appears as a promising neuroprotective molecule due to its pleiotropic action in the brain. Here, we investigated whether TAU plays a beneficial role in different behavioral domains of zebrafish submitted to an intermittent EtOH exposure model, specially focusing on social behavior, anxiety-like responses, and memory. Moreover, since monoamines play a role in EtOH-mediated responses, we also evaluated the influence of both TAU and EtOH exposures on brain monoamine oxidase (Z-MAO) activity. Fish were exposed to non-chlorinated water or 1% EtOH for 8 consecutive days (20 min per day). From the 5th day until the end of the experimental period (8th day), animals were kept in the absence or presence of TAU (42, 150, or 400 mg/L) 1 h per day immediately after EtOH exposure. Behavioral measurements started 24 h after the last EtOH exposure. We observed that TAU showed modest attenuating effects on shoaling behavior and anxiety-like responses, while 42 and 150 mg/L TAU abolished the memory acquisition deficit in the inhibitory avoidance task. Biochemical analysis revealed that TAU did not modulate EtOH-induced increase on brain Z-MAO activity. Collectively, our novel data show a potential beneficial effect of TAU in an intermittent EtOH exposure model in zebrafish. Moreover, these findings foster the growing utility of this aquatic species to investigate the neurobehavioral basis of EtOH- and TAU-mediated responses in vertebrates.

Acute effects of ethanol on behavioral responses of male and female zebrafish in the open field test with the influence of a non-familiar object.

In this report, we investigate whether the acute effects of different ethanol (EtOH) concentrations are sex-dependent in zebrafish subjected to the open field test (OFT) with the influence of a non-familiar object. Male and female zebrafish were separated into four groups and exposed to EtOH (0%, 0.25%, 0.5%, or 1.0% v/v) for 1 h. Fish were tested individually in the OFT and the tank was divided into three areas: periphery, intermediate, and center area. An object (black sphere; diameter: 1 cm) was placed in the center of the tank and behaviors were recorded for 5 min. At the baseline, females had a distinct exploratory activity and interaction pattern with the object, reflecting a more anxious and shyer behavior in relation to males. Females exposed to 0.5% EtOH performed more rapid investigation to the object than males, while 1.0% EtOH reduced locomotion in both sexes and increased immobility only in males. Principal component analyses revealed that anxiety-like behaviors, exploratory activity, and locomotion were the components that most accounted for total variances. Collectively, our novel findings show the existence of a sex-dependent effect in the zebrafish models acutely exposed to EtOH tested in the OFT with a non-familiar object.

Taurine prevents MK-801-induced shoal dispersion and altered cortisol responses in zebrafish.

Schizophrenia is a chronic neuropsychiatric disorder characterized by a shortened lifespan and significant impaired social and vocational functioning. Schizophrenic patients can present hypothalamic-pituitary-adrenal (HPA) axis dysfunctions and cortisol dysregulation, which play an important role on the etiology onset, exacerbation, and relapsing of symptoms. Based on its intrinsic neuroprotective properties, taurine is considered a promising substance with beneficial role on various brain disorders, including schizophrenia. Here, we evaluated the effects of taurine on shoaling behavior and whole-body cortisol levels in zebrafish treated with dizocilpine (MK-801), which elicits schizophrenia-like phenotypes in animal models. Briefly, zebrafish shoals (4 fish per shoal) were exposed to dechlorinated water or taurine (42, 150, or 400 mg/L) for 60 min. Then, saline (PBS, pH 7.4 or 2.0 mg/kg MK-801) were intraperitoneally injected and zebrafish behavior was recorded 15 min later. In general, MK-801 disrupted shoaling behavior and reduced whole-body cortisol levels in zebrafish. All taurine pretreatments prevented MK-801-induced increase in shoal area, while 400 mg/L taurine prevented the MK-801-induced alterations in neuroendocrine responses. Moreover, all taurine-pretreated groups showed increased geotaxis, supporting a modulatory role in the overall dispersion pattern of the shoal. Collectively, our novel findings show a potential protective effect of taurine on MK-801-induced shoal dispersion and altered neuroendocrine responses, fostering the use of zebrafish models to assess schizophrenia-like phenotypes.

Stress increases susceptibility to pentylenetetrazole-induced seizures in adult zebrafish.

Stress is the body's reaction to any change that requires adaptive responses. In various organisms, stress is a seizure-related comorbidity. Despite the exposure to stressors eliciting aversive behaviors in zebrafish, there are no data showing whether stress potentiates epileptic seizures in this species. Here, we investigated whether a previous exposure to an intense acute stressor positively modulates the susceptibility to seizures in pentylenetetrazole (PTZ)-challenged zebrafish. The conspecific alarm substance (CAS) was used to elicit aversive responses (3.5 mL/L for 5 min), observed by increased bottom dwelling and erratic movements. Then, fish were immediately exposed to 7.5 mM PTZ for 10 min to induce seizure-like behaviors. Stress increased the seizure intensity, the number of clonic-like seizure behaviors (score 4), as well as facilitated the occurrence of score 4 episodes by decreasing the latency in which fish reached the score 4. Moreover, fish with heightened anxiety showed increased susceptibility to PTZ, since positive correlations between anxiety- and seizure-like behaviors were found. Overall, since CAS also increased whole-body cortisol levels in zebrafish, our novel findings show a prominent response to PTZ-induced seizures in previously stressed zebrafish. Moreover, we reinforce the growing utility of zebrafish models to assess seizure-related comorbidities aiming to elucidate how stress can affect epileptic seizures in vertebrates.

Taurine-mediated aggression is abolished via 5-HT1A antagonism and serotonin depletion in zebrafish.

Taurine is one of the most abundant amino acids in vertebrates involved in important physiological functions, including osmoregulation, membrane stability, and neuronal activity. The pleiotropic effects of taurine support the existence of different mechanisms of action (e.g., modulation of GABAA, strychnine-sensitive glycine, and NMDA receptors), which can play a role in aggressive-related responses. However, the mechanisms underlying the effects of taurine on aggression are still poorly understood. Because aggression has been associated with diverse central mechanisms, especially serotonergic activity, we aimed to investigate the involvement of this system in taurine-induced aggression in zebrafish. We treated adult zebrafish with ρ-chlorophenylalanine (ρCPA), an inhibitor of the serotonin synthesis, as well as 5-HT1A receptor antagonist and agonist (WAY100135 and buspirone, respectively). Taurine effects were tested individually at three concentrations (42, 150, and 400 mg/L) for 60 min. We further analyzed the effects on aggression and locomotion using the mirror-induced aggression test. Taurine concentration that changed behavioral responses was selected to the succeeding pharmacological experiments using ρCPA, WAY100135, and buspirone. We found that buspirone did not alter the aggression. Yet, 42 mg/L taurine increased aggression, which was abolished by ρCPA and WAY100135, indicating the involvement of 5-HT1A receptors in taurine-mediated aggression. These set of data support an indirect mechanism mediating taurine-induced aggression via serotonin release and activation of 5-HT1A receptors in zebrafish. While the exact mechanisms underlying aggression are still unclear, our novel findings reveal a key role of the serotonergic system in the effects of taurine, supporting the use of zebrafish models to understand the neural basis of aggression in vertebrates.

Modulation of redox and insulin signaling underlie the anti-hyperglycemic and antioxidant effects of diphenyl diselenide in zebrafish.

The organic selenium compound diphenyl diselenide (DD) has been recognized as an antioxidant and neuroprotective agent, exerting an anti-hyperglycemic effect in experimental models of diabetes. However, the precise mechanisms involved in the protection are unclear. Using the zebrafish (Danio rerio) as a model organism, here we investigated biomarkers underlying the protective effects of DD against hyperglycemia, targeting in a transcriptional approach the redox and insulin-signaling pathway. Fish were fed on a diet containing DD (3 mg/kg) for 74 days. In the last 14 days, they were exposed to a 111 mM glucose solution to induce a hyperglycemic state. DD reduced blood glucose levels as well as normalized the brain mRNA transcription of four insulin receptors-coding genes (Insra1, Insra2, Insrb1, Insrb2), which were down-regulated by glucose. DD alone caused an up-regulation of relative mRNA transcription in both Insra receptors and glucose transporter 3 genes. DD counteracted hyperglycemia-induced lipid peroxidation, protein and thiol depletion. Along with the decreased activity of antioxidant enzymes SOD and GPx, the brain of hyperglycemic fish presented a reduction in mRNA transcription of FoxO3A, FoxO3B, Nrf2, GPx3A, SOD1, and SOD2 genes. Besides normalizing the transcriptional levels, DD caused an up-regulation of relative mRNAs that encode Nrf2, FoxO1A, FOXO3A, GPx4A, PTP1B, AKT and SelP. Collectively, our findings suggest that the antioxidant and anti-hyperglycemic actions of DD in a zebrafish diabetes model are likely associated with the regulation of the oxidative stress resistance and the insulin-signaling pathway and that could be related to the modulation at mRNA level of two important transcription factors, Nrf2 and FoxO.

Neuroprotective role of taurine on MK-801-induced memory impairment and hyperlocomotion in zebrafish.

Schizophrenia is a neuropsychiatric condition that reaches around 1% of people worldwide. Because taurine exerts a neuroprotective role in the brain, this molecule is a promising candidate to reduce schizophrenia-like symptoms. Here, we investigated a possible neuroprotective role of taurine against MK-801-induced memory deficit and hyperlocomotion in zebrafish using the inhibitory avoidance task and the novel tank diving test, respectively. First, we assessed the influence of different MK-801 doses (0.1, 0.3, 0.5, 1 and 2 mg/kg, i.p.) on memory consolidation. Although all MK-801 doses tend to reduce the retention index, only 2 mg/kg MK-801 showed robust amnesic effects. Then, we evaluated whether taurine pretreatments (42, 150 and 400 mg/L for 60 min) prevent MK-801-induced cognitive impairment. Immediately after the training, animals were exposed to non-chlorinated water or taurine and subsequently challenged with 2 mg/kg MK-801, i.p. The test session was performed 24 h after training. Although taurine alone did not change memory retention when compared with control, taurine pretreatments prevented MK-801-induced memory deficit. Importantly, no locomotor changes were observed 24 h after the training session. In the novel tank diving test, MK-801 induced hyperlocomotion and disrupted vertical activity, while 400 mg/L taurine pretreatment prevented these effects. Overall, our novel findings indicate a neuroprotective role of taurine against MK-801-induced memory deficit and hyperlocomotion, reinforcing the growing utility of zebrafish models to investigate the beneficial effects of different compounds against glutamate excitotoxicity.

Understanding the neurobiological effects of drug abuse: Lessons from zebrafish models.

Drug abuse and brain disorders related to drug comsumption are public health problems with harmful individual and social consequences. The identification of therapeutic targets and precise pharmacological treatments to these neuropsychiatric conditions associated with drug abuse are urgently needed. Understanding the link between neurobiological mechanisms and behavior is a key aspect of elucidating drug abuse-related targets. Due to various molecular, biochemical, pharmacological, and physiological features, the zebrafish (Danio rerio) has been considered a suitable vertebrate for modeling complex processes involved in drug abuse responses. In this review, we discuss how the zebrafish has been successfully used for modeling neurobehavioral phenotypes related to drug abuse and review the effects of opioids, cannabinoids, alcohol, nicotine, and psychedelic drugs on the central nervous system (CNS). Moreover, we summarize recent advances in zebrafish-based studies and outline potential advantages and limitations of the existing zebrafish models to explore the neurochemical bases of drug abuse and addiction. Finally, we discuss how the use of zebrafish models may present fruitful approaches to provide valuable clinically translatable data.

Role of the serotonergic system in ethanol-induced aggression and anxiety: A pharmacological approach using the zebrafish model.

Acute ethanol (EtOH) consumption exerts a biphasic effect on behavior and increases serotonin levels in the brain. However, the molecular mechanisms underlying alcohol-mediated behavioral responses still remain to be fully elucidated. Here, we investigate pharmacologically the involvement of the serotonergic pathway on acute EtOH-induced behavioral changes in zebrafish. We exposed zebrafish to 0.25, 0.5, 1.0% (v/v) EtOH for 1 h and analyzed the effects on aggression, anxiety-like behaviors, and locomotion. EtOH concentrations that changed behavioral responses were selected to the subsequent experiments. As a pharmacological approach, we used pCPA (inhibitor of tryptophan hydroxylase), WAY100135 (5-HT1A antagonist), buspirone (5-HT1A agonist), CGS12066A and CGS12066B (5-HT1B antagonist and agonist, respectively), ketanserin (5-HT2A antagonist) and (±)-DOI hydrochloride (5-HT2A agonist). All serotonergic receptors tested modulated aggression, with a key role of 5-HT2A in aggressive behavior following 0.25% EtOH exposure. Because CGS12066B mimicked 0.5% EtOH anxiolysis, which was antagonized by CGS12066A, we hypothesized that anxiolytic-like responses are possibly mediated by 5-HT1B receptors. Conversely, the depressant effects of EtOH are probably not related with direct changes on serotonergic pathway. Overall, our novel findings demonstrate a role of the serotonergic system in modulating the behavioral effects of EtOH in zebrafish. These data also reinforce the growing utility of zebrafish models in alcohol research and help elucidate the neurobiological mechanisms underlying alcohol abuse and associated complex behavioral phenotypes.

Concomitant taurine exposure counteracts ethanol-induced changes in locomotor and anxiety-like responses in zebrafish.

Taurine (TAU) is a ß-amino sulfonic acid with pleiotropic roles in the brain, including the neuromodulatory activity via GABAergic and glycinergic agonism. This molecule is found at high concentrations in energy drinks and is often mixed with alcohol in beverages. Although TAU has a neuroprotective role in the brain, the putative risks of mixing TAU and EtOH are not fully understood. Here, we investigated whether TAU modulates locomotor and anxiety-like behavior in adult zebrafish by using the novel tank and light-dark tests following acute EtOH exposure at anxiogenic and anxiolytic concentrations. Zebrafish were individually exposed to water (control), TAU (42, 150, and 400 mg/L), and EtOH (0.25% (v/v) and 1% (v/v)) both independently and cotreated for 1 h. EtOH 0.25% and TAU produced U-shape anxiolytic-like behavior in the light-dark test, TAU 42 and 400 positively modulated EtOH effects, and TAU 150 exerted a protective effect. All TAU concentrations counteracted EtOH 1%-induced locomotion impairment, as well as the anxiogenic-like behavior. Finally, all TAU concentrations when given independently or cotreated with EtOH 0.25% and 1% decreased the risk assessment of the lit compartment. Principal component analyses revealed that exploration and anxiety-like responses were the main behaviors that contribute to the effects of TAU and EtOH. Overall, we demonstrate that TAU differently modulates EtOH-induced anxiolytic- and anxiogenic-like behaviors depending on the concentration, suggesting a complex mechanism underlying TAU and EtOH interactions.

Neurochemical mechanisms underlying acute and chronic ethanol-mediated responses in zebrafish: The role of mitochondrial bioenergetics.

Ethanol (EtOH) is a socially-accepted drug, whose consumption is a risk factor for non-intentional injuries, development of pathologies, and addiction. In the brain, EtOH affects redox signaling and increases reactive oxygen species (ROS) production after acute and chronic exposures. Here, using a high-resolution respirometry assay, we investigated whether changes in mitochondrial bioenergetics play a role in both acute and chronic EtOH-mediated neurochemical responses in zebrafish. For the first time, we showed that acute and chronic EtOH exposures differently affect brain mitochondrial function. Acutely, EtOH stimulated mitochondrial respiration through increased baseline state, CI-mediated OXPHOS, OXPHOS capacity, OXPHOS coupling efficiency, bioenergetic efficiency, and ROX/ETS ratio. Conversely, EtOH chronically decreased baseline respiration, complex I- and II-mediated ETS, as well as increased ROX state and ROX/ETS ratio, which are associated with ROS formation. Overall, we observed that changes in mitochondrial bioenergetics play a role, at least partially, in both acute and chronic effects of EtOH in the zebrafish brain. Moreover, our findings reinforce the face, predictive, and construct validities of zebrafish models to explore the neurochemical bases involved in alcohol abuse and alcoholism.

Modeling psychiatric comorbid symptoms of epileptic seizures in zebrafish.

Epilepsy is a debilitating neurological disorder characterized by recurrent unprovoked seizures. Anxiety, cognitive deficits, depressive-like symptoms, and social dysfunction are psychiatric comorbidities with high prevalence in epileptic patients. Due to the genetic and behavioral tractability, the zebrafish is a promising model organism to understand the neural bases involved in epilepsy-related comorbidities. Here, we aimed to characterize some behavioral phenotypes paralleling those observed in epilepsy-related comorbidities after a single pentylenetetrazole (PTZ) exposure in zebrafish. We also analyzed the influence of whole-body cortisol levels in the behavioral responses measured. Fish were exposed to 10 mM PTZ for 20 min to induce epileptic seizures. After 24 h recovery period, locomotion and anxiety-like responses (novel tank and light-dark tests), social interaction (shoaling behavior task), and memory retention (inhibitory avoidance protocol) were assessed. Basically, PTZ impaired habituation to novelty stress, evoked anxiogenic-like behaviors, disrupted shoaling, and caused memory consolidation deficits in zebrafish without changing whole-body cortisol levels. In conclusion, our novel findings further validate the use of zebrafish as a suitable tool for modeling epilepsy-related comorbidities in translational neuropsychiatric research.

Involvement of anxiety-like behaviors and brain oxidative stress in the chronic effects of alarm reaction in zebrafish populations.

Aversive conditions elicit anxiety responses that prepare the organism to an eventual threat. Nonetheless, prolonged anxiety is a pathological condition associated with various neuropsychiatric disorders. Here, we evaluated whether the conspecific alarm substance (CAS), a chemical cue that elicits aversion, influences anxiety-like behaviors and modulates brain oxidative stress-related parameters in wild-type (WT) and leopard (leo) zebrafish following a repeated exposure protocol. CAS exposure was performed for 5 min, once daily for 7 consecutive days. In the 8th day, animals were tested in the light/dark and novel tank tests and their brains were further dissected for biochemical analyses. CAS chronically induced anxiogenic-like states in WT and leo populations when their behaviors were analyzed in the light/dark and novel tank tests. CAS also increased catalase (CAT) and glutathione S-transferase (GST) activities, as well as non-protein thiol (NPSH) content in WT and leo, but only leo had increased thiobarbituric reactive substance (TBARS) levels in the brain. At baseline conditions, leo was more 'anxious' when compared to WT, displaying lower CAT activity and carbonylated protein (CP) levels. Overall, CAS chronically triggers anxiety-like behavior in zebrafish populations, which may be associated with changes in oxidative stress-related parameters. Furthermore, the use of different zebrafish populations may serve as an interesting tool in future research aiming to investigate the neurobehavioral bases of neuropsychiatric disorders in vertebrates.

Exploring Object Discrimination in Zebrafish: Behavioral Performance and Scopolamine-Induced Cognitive Deficits at Different Retention Intervals.

The object discrimination test allows the testing of different memory retention periods. However, few behavioral endpoints have been measured in fish species such that retention is often assessed using a single parameter (time spent in object area). Here, we aimed to explore the object discrimination test in zebrafish by assessing their behavioral performance after 1 or 24 h retention interval periods. To characterize putative interaction-like behaviors, fish were tested in the absence or presence of scopolamine (1 h before test session). Zebrafish were habituated for 3 consecutive days in the experimental tank, and training session was performed for 10 min using two identical nonpreferred objects (black cube or sphere). After the retention intervals, a familiar object was replaced by a novel object (test session, 10 min). Fish were also exposed to the novel tank diving test to assess locomotion and anxiety-like behaviors. At 1 h retention interval, animals performed more circular-like investigation near the familiar object, whereas 24 h after training session, a prominent rapid investigation was observed when animals explore the nonfamiliar object. Because scopolamine abolished these phenotypes, as well as the increased time spent in the novel object area during the test without changing locomotion and anxiety-related parameters, the behavioral responses described here may predictively reflect interaction-like behaviors involved in object discrimination memory in zebrafish models.

Nicotine prevents anxiety-like behavioral responses in zebrafish.

Anxiety-related disorders are severe psychiatric conditions that involve complex physiological and behavioral maladaptive responses. The use of conspecific alarm substance (CAS) for inducing anxiety-like behaviors in fish species provides important translational insights of how aversive conditions modulate neurobehavioral functions. Because nicotine may elicit anxiolytic-like responses, here we investigated whether acute nicotine exposure prevents CAS-induced anxiogenic-like behaviors in zebrafish. We used both novel tank and light-dark tests as two well-established paradigms for measuring anxiety-like phenotypes. Fish were individually exposed to 1 mg/L nicotine or non-chlorinated water for 3 min and then transferred to other tanks in the absence or presence of 3.5 mL/L CAS for 5 min. Later, the behavior of fish was tested in the novel tank test or in the light-dark preference test. As expected, CAS triggered aversive behaviors by increasing bottom-dwelling, freezing, erratic movements, scototaxis, and risk assessment episodes. Nicotine alone elicited anxiolytic-like behaviors since it increased the time spent in the top, as well as the average duration of entry in the lit compartment. Moreover, nicotine pretreatment prevented CAS-induced aversive responses without changing locomotion, suggesting that anxiolysis could play a role, at least in part, to the behavioral effects of nicotine observed here. Overall, these novel findings show the beneficial effects of nicotine on anxiogenic responses in zebrafish. We also reinforce the practical advantages of this aquatic species to explore the relieving properties of nicotine, as well as to understand the neurobiological bases involved in anxiety-related disorders and associated therapeutic targets.

Taurine prevents memory consolidation deficits in a novel alcohol-induced blackout model in zebrafish.

Ethanol is one of the most consumed substance worldwide that impairs learning and memory processes, resulting in amnesia or blackout. Due to the genetic conservation, rich behavioral repertoire, and high pharmacological tractability, the zebrafish (Danio rerio) has emerged as a powerful model organism for assessing preventive strategies against the noxious effects of ethanol in vertebrates. Here, we used an inhibitory avoidance apparatus to investigate the potential preventive effects of taurine in a novel ethanol-induced amnesia model in zebrafish. The experimental tank consisted of two compartments of the same size, one dark and another white, which were separated by a guillotine-type door. Three parallel metal bars coupled to an electrical stimulator were connected on each lateral wall of the dark compartment as electrical stimulus source. Differences on the latency to enter the dark compartment were used as retention indexes. A mild electric shock (125 mA, 3 ±â€¯0.2 V) at 10 and 1000 Hz did not promote significant learning, while 100 Hz facilitated memory retention. Posttraining administration of MK-801 blocked this response, reinforcing the predictive validity of the test. Treatments were performed immediately after the training session using the 100 Hz frequency. Animals were exposed to water (control), taurine (42, 150, 400 mg/L), ethanol (0.25%, 1.0% v/v) or taurine plus ethanol to assess the effects on memory consolidation. Test session was performed 24 h following training. Ethanol at 0.25% did not affect memory consolidation, but 1.0% impaired memory without changing locomotion. Although taurine alone did not modulate learning, all concentrations tested exerted prevented ethanol-induced memory impairment. Overall, we describe a novel ethanol-induced blackout model, where a high ethanol concentration acutely impairs memory consolidation in zebrafish. Moreover, since taurine showed a protective role, we reinforce the growing utility of zebrafish models for assessing the deleterious effects of ethanol and potential therapeutic strategies.