A proposed correction in the weighted method to score the Ishihara test.

OBJECTIVE: Ishihara test is a color vision test, whose results consider that all plates of the test have the same weighting. Rodriguez-Carmona et al. (Aviat Space Environ Med 83:19-29, 2012) proposed an equation to quantify the Ishihara test results (severity index), which took an account the rate of hits from the different plates of the test considering the performance of trichromat or colorblind population. We proposed a correction in Rodiguez-Carmona's equation for the severity index. We evaluated 60 normal trichromats and 107 subjects with congenital color deficiency. We calculated three indexes to quantify the results of each subject: a non-weighted index, a weighted index similar to the Rodriguez-Carmona et al., and a weighted index modified which combined the hit frequency for each plate in a trichromat population and of the error reading frequency for each plate in color-blind populations. RESULTS: Compared to the non-weighted evaluation, the weighted index was reduced by 22.95%, 32.92%, and 35.38% from trichromats, protan and deutan groups, respectively. Receiver Operating Characteristics (ROC) analysis showed perfect performance of the classifier for all metrics to measure the Ishihara test results. The proposal correction changed significantly the value of the index, but the overall benefits were small.

Evidence of Asymptomatic Visual Losses after Surgical Repair of Cerebral Aneurysm.

Deficits in visual acuity, visual field, and oculomotor function are commonly detected after repair of cerebral aneurysms. However, when these deficits are absent, it does not mean that other potential visual deficits also are absent. Here, we report three cases that after complete recover from surgical repair of cerebral aneurysms presented minimal visual acuities of about 20/20 and no visual disturbances. While two of them (Cases 1 and 2) showed visual fields with no relevant central defects, two of them showed relevant impairments in spatial contrast sensitivity (Cases 2 and 3). This evidence supports that after complete recover from surgical repair of hemorrhagic cerebral aneurysms spatial contrast sensitivity can be asymptomatically impaired when visual acuity (Cases 2 and 3) and visual fields (Case 2) are not correlated with symptoms of visual disturbances. Hypothetical explanations and consequences of such evidence are discussed.

Comparative Analyses of Zebrafish Anxiety-Like Behavior Using Conflict-Based Novelty Tests.

Modeling of stress and anxiety in adult zebrafish (Danio rerio) is increasingly utilized in neuroscience research and central nervous system (CNS) drug discovery. Representing the most commonly used zebrafish anxiety models, the novel tank test (NTT) focuses on zebrafish diving in response to potentially threatening stimuli, whereas the light-dark test (LDT) is based on fish scototaxis (innate preference for dark vs. bright areas). Here, we systematically evaluate the utility of these two tests, combining meta-analyses of published literature with comparative in vivo behavioral and whole-body endocrine (cortisol) testing. Overall, the NTT and LDT behaviors demonstrate a generally good cross-test correlation in vivo, whereas meta-analyses of published literature show that both tests have similar sensitivity to zebrafish anxiety-like states. Finally, NTT evokes higher levels of cortisol, likely representing a more stressful procedure than LDT. Collectively, our study reappraises NTT and LDT for studying anxiety-like states in zebrafish, and emphasizes their developing utility for neurobehavioral research. These findings can help optimize drug screening procedures by choosing more appropriate models for testing anxiolytic or anxiogenic drugs.

Time-dependent sensitization of stress responses in zebrafish: A putative model for post-traumatic stress disorder.

Time-dependent sensitization (TDS)-the delayed increase in neurobehavioral responses to heterotypic stressors after exposure to an intense, inescapable stressor-has been proposed as an animal model for post-traumatic stress disorder (PTSD). Translationally relevant stressors used in TDS are capable of affecting more than one behavioral domain and produce interindividual variability in responsiveness. Here, conspecific alarm substance (CAS) is shown to induce TDS in zebrafish in inter- and intra-population-specific way. Exposure to CAS, an ecologically relevant stimulus which produces fear-like responses acutely, increased anxiety and arousal in zebrafish from the blue shortfin (BSF) phenotype 24h after stimulus delivery. Anxiety-like responses were differently affected immediately and 24h after stimulus delivery. Anxiety-like responses were more sensitized in zebrafish from the longfin (LOF) than in the BSF phenotype, an effect which is reminiscent of "basal" differences in anxiety-like behavior. After application of behavioral cutoff criteria, CAS was shown to produce intense TDS in ∼25% of LOF animals, while ∼20% of exposed animals showed little evidence of TDS. Overall, these results suggest that CAS induces TDS in zebrafish after a 24h "incubation" period, with inter- and intra-population variability that underlines its face and ecological validity.

Non-mammalian models in behavioral neuroscience: consequences for biological psychiatry.

Current models in biological psychiatry focus on a handful of model species, and the majority of work relies on data generated in rodents. However, in the same sense that a comparative approach to neuroanatomy allows for the identification of patterns of brain organization, the inclusion of other species and an adoption of comparative viewpoints in behavioral neuroscience could also lead to increases in knowledge relevant to biological psychiatry. Specifically, this approach could help to identify conserved features of brain structure and behavior, as well as to understand how variation in gene expression or developmental trajectories relates to variation in brain and behavior pertinent to psychiatric disorders. To achieve this goal, the current focus on mammalian species must be expanded to include other species, including non-mammalian taxa. In this article, we review behavioral neuroscientific experiments in non-mammalian species, including traditional "model organisms" (zebrafish and Drosophila) as well as in other species which can be used as "reference." The application of these domains in biological psychiatry and their translational relevance is considered.

Putative involvement of the nitrergic system on the consolidation, but not initiation, of behavioral sensitization after conspecific alarm substance in zebrafish.

Stressful manipulations can sensitize the behavior of an organism, increasing anxiety-like behavior after a delay; this long-term stress sensitization can represent the pathophysiological basis of trauma- and stress-related disorders (TRSDs), of which the most prevalent is post-traumatic stress disorder (PTSD). A role for the glutamate-nitric oxide pathway in this sensitization is implied by behavioral, neurophysiological and genomic data on different species. Here, we report on the long-term sensitization of anxiety-like behavior in zebrafish and the possible participation of nitric oxide in this process. Zebrafish exposed to a conspecific alarm substance (AS) show increased anxiety-like behavior at least 24h after stimulus delivery. Blocking nitric oxide synthesis with l-NAME (5mg/kg) 30min, but not 90min, after AS exposure blocks the sensitization of scototaxis and risk assessment, while treatment 90min after exposure blocks the sensitization of thigmotaxis and erratic swimming; l-NAME was not effective when administered 30min before AS exposure. These data suggest a participation of nitric oxide in the consolidation, but not in the initiation, of behavioral sensitization after predator threat.

Role of nitric oxide in the behavioral and neurochemical effects of IB-MECA in zebrafish.

RATIONALE: The adenosine A3 receptor and the nitric oxide (NO) pathway regulate the function and localization of serotonin transporters (SERTs). These transporters regulate extracellular serotonin levels, which are correlated with defensive behavior. OBJECTIVE: The purpose of this study was to understand the role of the A3AR on anxiety and arousal models in zebrafish, and whether this role is mediated by the nitrergic modulation of serotonin uptake. METHODS: The effects of IB-MECA (0.01 and 0.1 mg/kg) were assessed in a series of behavioral tasks in adult zebrafish, as well as on extracellular serotonin levels in vivo and serotonin uptake in brain homogenates. Finally, the interaction between IB-MECA and drugs blocking voltage-dependent calcium channels (VDCCs), NO synthase, and SERT was analyzed. RESULTS: At the lowest dose, IB-MECA decreased bottom dwelling and scototaxis, while at the highest dose, it also decreased shoaling, startle probability, and melanophore responses. These effects were accompanied by an increase in brain extracellular serotonin levels. IB-MECA also concentration-dependently increased serotonin uptake in vitro. The effects of IB-MECA on extracellular 5-HT, scototaxis, and geotaxis were blocked by L-NAME, while only the effects on 5-HT and scototaxis were blocked by verapamil. In vitro, the increase in 5-HT uptake was dependent on VDCCs and NO. Finally, fluoxetine blocked the effect of IB-MECA on scototaxis, but not geotaxis. CONCLUSION: These results suggest that the effect of IB-MECA on scototaxis are mediated by a VDCC-NO-SERT pathway. While NO seems to mediate the effects of IB-MECA on geotaxis, neither VDCCs nor SERT seems to be involved in this process.

Interactions between serotonin and glutamate-nitric oxide pathways in zebrafish scototaxis.

NMDA receptors have been implicated in the acute response to stress, possibly mediated the nitric oxide pathway; serotonin has also been implicated in these responses, and has recently been shown to modulate the nitric oxide pathway via 5-HT1 and 5-HT2 receptors. In this work, we compare the effects of NMDA and a 5-HT1A receptor ligands on light/dark preference in adult zebrafish, and investigate whether nitric oxide mediates the effects of such drugs. The noncompetitive NMDA receptor antagonist MK-801 decreased dark preference (scototaxis), while NMDA increased it; the effects of NMDA were completely blocked by pretreatment with the nitric oxide synthase (NOS) antagonist L-NAME. SNP, a nitric oxide donor, produced a bell-shaped dose-response profile on scototaxis. Treatment with 5-HTP increased scototaxis, an effect which was potentiated by pre-treatment with NMDA, but not MK-801, and partially blocked by L-NAME. The 5-HT1A receptor antagonist WAY 100,635 decreased scototaxis, an effect which was completely blocked by L-NAME. These results suggest that tonic NOS inhibition is an important downstream effector of 5-HT1A receptors in the regulation of dark preference behavior in zebrafish, and that NOS is also under phasic independent control by NMDA receptors.

Interaction between 5-HT1B receptors and nitric oxide in zebrafish responses to novelty.

Nitric oxide (NO) and serotonin (5-HT) interact at the molecular and systems levels to control behavioral variables, including agression, fear, and reactions to novelty. In zebrafish, the 5-HT1B receptor has been implicated in anxiety and reactions to novelty, while the 5-HT1A receptor is associated with anxiety-like behavior; this role of the 5-HT1A receptor is mediated by NO. This work investigated whether NO also participates in the mediation of novelty responses by the 5-HT1B receptor. The 5-HT1B receptor inverse agonist SB 224,289 decreased bottom-dwelling and erratic swimming in zebrafish; the effects on bottom-dwelling, but not on erratic swimming, were blocked by pre-treatment with the nitric oxide synthase inhibitor L-NAME. These effects underline a novel mechanism by which 5-HT controls zebrafish reactivity to novel environments, with implications for the study of neotic reactions, exploratory behavior, and anxiety-like states.

Fingerprinting of psychoactive drugs in zebrafish anxiety-like behaviors.

A major hindrance for the development of psychiatric drugs is the prediction of how treatments can alter complex behaviors in assays which have good throughput and physiological complexity. Here we report the development of a medium-throughput screen for drugs which alter anxiety-like behavior in adult zebrafish. The observed phenotypes were clustered according to shared behavioral effects. This barcoding procedure revealed conserved functions of anxiolytic, anxiogenic and psychomotor stimulating drugs and predicted effects of poorly characterized compounds on anxiety. Moreover, anxiolytic drugs all decreased, while anxiogenic drugs increased, serotonin turnover. These results underscore the power of behavioral profiling in adult zebrafish as an approach which combines throughput and physiological complexity in the pharmacological dissection of complex behaviors.

Fluoxetine and WAY 100,635 dissociate increases in scototaxis and analgesia induced by conspecific alarm substance in zebrafish (Danio rerio Hamilton 1822).

Alarm reactions to a substance secreted by the damaged skin of conspecifics and closely-related species are increasingly being recognized as fear-like responses in fish. The neurochemical underpinnings of these effects are so far unknown; however, given the role of the serotonergic system on defensive behavior, it is possible that the alarm reaction is mediated by this monoamine. Exposure to conspecific alarm substance (CAS) increased anxiety-like behavior in the light/dark test in zebrafish and decreased nocifensive behavior. These effects were accompanied by increases in blood glucose, hemoglobin, epinephrine and norepinephrine levels, as well as extracellular levels of serotonin in the brain. Pretreatment with fluoxetine blocked the anxiogenic effects of CAS on the light/dark test as well as all physiological parameters and the increase in extracellular brain 5-HT, but not the reduction in nocifensive behavior. Conversely, pretreatment with the 5-HT1AR antagonist WAY 100635 blocked the effects on nocifensive behavior, but not the effects on anxiety-like behavior nor on physiological parameters. These results point to an important and complex role of the serotonergic system in the mediation of fear-potentiated behavior in the light/dark test and in fear-induced analgesia in zebrafish.

Nitric oxide as a regulatory molecule in the processing of the visual stimulus.

Nitric oxide (NO) is a highly reactive gas with considerable diffusion power that is produced pre- and post synaptically in the central nervous system (CNS). In the visual system, it is involved in the processing of the visual information from the retina to superior visual centers. In this review we discuss the main mechanisms through which nitric oxide acts, in physiological levels, on the retina, lateral geniculate nucleus (LGN) and primary visual cortex. In the retina, the cGMP-dependent nitric oxide activity initially amplifies the signal, subsequently increasing the inhibitory activity, suggesting that the signal is "filtered". In the thalamus, on dLGN, neuronal activity is amplified by NO derived from brainstem cholinergic cells, in a cGMP-independent mechanism; the result is the amplification of the signal arriving from retina. Finally, on the visual cortex (V1), NO acts through changes on the cGMP levels, increasing signal detection. These observations suggest that NO works like a filter, modulating the signal along the visual pathways.

Role of serotonin in zebrafish (Danio rerio) anxiety: relationship with serotonin levels and effect of buspirone, WAY 100635, SB 224289, fluoxetine and para-chlorophenylalanine (pCPA) in two behavioral models.

Serotonin (5-HT) is a neurotransmitter that is involved in many behavioral functions, including the organization of defense, and its putative pathological correlate, anxiety and stress disorders. Recently, behavioral tests for anxiety have been proposed in zebrafish. Exposure to the novel tank test or to the light/dark test increased extracellular fluid 5-HT content in the brain; anxiety-like behavior correlated positively with 5-HT content in the novel tank test, while the correlation was negative in the light/dark test. Acute treatment with a low dose of fluoxetine was anxiolytic in the geotaxis test and anxiogenic in the scototaxis test, while treatment with a higher dose produced a hyperlocomotor effect in both tasks. Buspirone and WAY 100635 were anxiolytic in both tests, while SB 224289 was anxiolytic in the geotaxis and slightly anxiogenic in the scototaxis test. Serotonin depletion with pCPA was anxiogenic in the geotaxis and anxiolytic in scototaxis. These results underline the differential sensitivity of these tasks to assess serotonergic agents; alternatively, serotonin might regulate zebrafish behavior differently in the novel tank test and in the light/dark test.

"Limbic associative" and "autonomic" amygdala in teleosts: a review of the evidence.

The amygdaloid nuclei form an important hub of structures associated with diverse aspects of cognition and emotional behavior. Homologous structures have been determined in tetrapods, but homology of amygdala-like regions in bony fishes is presently unclear. Based on connectivity patterns, genoarchitecture, chemical neuroanatomy, and functional studies, we suggest that the dorsomedial portion of the pallium of Actinopterygii is the homolog of the basolateral/lateral amygdala ("frontotemporal amygdaloid system"), while the supracommissural and postcommissural portions of the subpallium are homologous to the extended central amygdala (central amygdaloid nucleus and bed nucleus of the stria terminalis). Nonetheless, the differentiation between these nuclei is not as clear-cut as in mammals, and there is no clear evidence for the existence of an "olfactory" medial amygdala in Actinopterygii, suggesting that the parcellation of one or two amygdaloid nuclei into many subnuclei occurred with the appearance of a true vomeronasal system.

Psychophysical evaluation of achromatic and chromatic vision of workers chronically exposed to organic solvents.

The purpose of this paper was to evaluate achromatic and chromatic vision of workers chronically exposed to organic solvents through psychophysical methods. Thirty-one gas station workers (31.5 ± 8.4 years old) were evaluated. Psychophysical tests were achromatic tests (Snellen chart, spatial and temporal contrast sensitivity, and visual perimetry) and chromatic tests (Ishihara's test, color discrimination ellipses, and Farnsworth-Munsell 100 hue test--FM100). Spatial contrast sensitivities of exposed workers were lower than the control at spatial frequencies of 20 and 30 cpd whilst the temporal contrast sensitivity was preserved. Visual field losses were found in 10-30 degrees of eccentricity in the solvent exposed workers. The exposed workers group had higher error values of FM100 and wider color discrimination ellipses area compared to the controls. Workers occupationally exposed to organic solvents had abnormal visual functions, mainly color vision losses and visual field constriction.

Possible role of serotoninergic system in the neurobehavioral impairment induced by acute methylmercury exposure in zebrafish (Danio rerio).

Adult zebrafish were treated acutely with methylmercury (1.0 or 5.0 µg g(-1), i.p.) and, 24h after treatment, were tested in two behavioral models of anxiety, the novel tank and the light/dark preference tests. At the smaller dose, methylmercury produced a marked anxiogenic profile in both tests, while the greater dose produced hyperlocomotion in the novel tank test. These effects were accompanied by a decrease in extracellular levels of serotonin, and an increase in extracellular levels of tryptamine-4,5-dione, a partially oxidized metabolite of serotonin. A marked increase in the formation of malondialdehyde, a marker of oxidative stress, accompanied these parameters. It is suggested that methylmercury-induced oxidative stress produced mitochondrial dysfunction and originated tryptamine-4,5-dione, which could have further inhibited tryptophan hydroxylase. These results underscore the importance of assessing acute, low-level neurobehavioral effects of methylmercury.

Métodos utilizados na avaliação psicofísica da visão de cores humana / Methods used in the psychophysics assessment of human color / Méthodes utilisées dans l'évaluation psychophysique de la vision humaine des couleurs / Métodos usados en la evaluación psicofísica de de la visión de colores humana

A cor é um atributo perceptual que nos permite identificar e localizar padrões ambientais de mesmo brilho e constitui uma dimensão adicional na identificação de objetos, além da detecção de inúmeros outros atributos dos objetos em sua relação com a cena visual, como luminância, contraste, forma, movimento, textura, profundidade. Decorre daí a sua importância fundamental nas atividades desempenhadas pelos animais e pelos seres humanos em sua interação com o ambiente. A psicofísica visual preocupa-se com o estudo quantitativo da relação entre eventos físicos de estimulação sensorial e a resposta comportamental resultante desta estimulação, fornecendo dessa maneira meios de avaliar aspectos da visão humana, como a visão de cores. Este artigo tem o objetivo de mostrar diversas técnicas eficientes na avaliação da visão cromática humana através de métodos psicofísicos adaptativos(AU)

Color is a perceptual attribute that allows organisms to identify and to locate environmental patterns of equal brightnesses and constitutes an additional dimension in object identification, in addition to the detection of several other object dimensions in relation with the visual scene. Color therefore serves an important role in animal and human interaction with the environment. By supplying ways to evaluate aspects of human vision, including color vision, visual psychophysics focusses on the quantitative study of the relation between physical events of sensory stimulation and the resulting behavioral response. The objective of this paper is to demonstrate several efficient techniques in the evaluation of the chromatic human vision through adaptive psychophysical methods(AU)

Une couleur est un attribut perceptuel qui permet aux organismes d'identifier et de localiser des normes ambiantes de même luminosité. Les couleurs servant d'identification dimensionnelle des objets, celles-ci sont très importantes dans l'activité quotidienne des animaux, et en particulier les humains. La psychophysique visuelle se se concentre sur l'étude quantitative de la relation entre les événements physiques de stimulation sensorielle et la réponse comportemental résultant de cette stimulation pour évaluer certains aspects de la vision humaine, comme la perception chromatique. L'objectif de ce texte est de démontrer les diverses techniques efficaces pour l'évaluation de la vision chromatique humaine en utilisant des méthodes psychophysiques adaptatives(AU)

El color es una cualidad perceptual que nos permite identificar y localizar estándares ambientales de mismo brillo y constituye una dimensión adicional en la identificación de los objetos, además de la detección de inúmeras otras cualidades de los objetos en su relación con la escena visual, como luminancia, contraste, forma, movimiento, textura, profundidad. Ahí está su importancia básica en las actividades hechas por los animales y por los seres humanos en su interacción con el ambiente. La psicofísica visual está preocupada del estudio cuantitativo de la relación entre los acontecimientos físicos de estimulación sensorial y la respuesta de comportamiento resultante de esta estimulación, suministrando de esta forma medios de evaluar aspectos de la visión humana, como la visión de colores. Este manuscrito tiene como objetivo demostrar diversas técnicas eficientes en la evaluación de la visión cromática humana a través de métodos psicofísicos adaptativos(AU)

Métodos utilizados na avaliação psicofísica da visão de cores humana / Methods used in the psychophysics assessment of human color / Méthodes utilisées dans l'évaluation psychophysique de la vision humaine des couleurs / Métodos usados en la evaluación psicofísica de de la visión de colores humana

A cor é um atributo perceptual que nos permite identificar e localizar padrões ambientais de mesmo brilho e constitui uma dimensão adicional na identificação de objetos, além da detecção de inúmeros outros atributos dos objetos em sua relação com a cena visual, como luminância, contraste, forma, movimento, textura, profundidade. Decorre daí a sua importância fundamental nas atividades desempenhadas pelos animais e pelos seres humanos em sua interação com o ambiente. A psicofísica visual preocupa-se com o estudo quantitativo da relação entre eventos físicos de estimulação sensorial e a resposta comportamental resultante desta estimulação, fornecendo dessa maneira meios de avaliar aspectos da visão humana, como a visão de cores. Este artigo tem o objetivo de mostrar diversas técnicas eficientes na avaliação da visão cromática humana através de métodos psicofísicos adaptativos.

Color is a perceptual attribute that allows organisms to identify and to locate environmental patterns of equal brightnesses and constitutes an additional dimension in object identification, in addition to the detection of several other object dimensions in relation with the visual scene. Color therefore serves an important role in animal and human interaction with the environment. By supplying ways to evaluate aspects of human vision, including color vision, visual psychophysics focusses on the quantitative study of the relation between physical events of sensory stimulation and the resulting behavioral response. The objective of this paper is to demonstrate several efficient techniques in the evaluation of the chromatic human vision through adaptive psychophysical methods.

Une couleur est un attribut perceptuel qui permet aux organismes d'identifier et de localiser des normes ambiantes de même luminosité. Les couleurs servant d'identification dimensionnelle des objets, celles-ci sont très importantes dans l'activité quotidienne des animaux, et en particulier les humains. La psychophysique visuelle se se concentre sur l'étude quantitative de la relation entre les événements physiques de stimulation sensorielle et la réponse comportemental résultant de cette stimulation pour évaluer certains aspects de la vision humaine, comme la perception chromatique. L'objectif de ce texte est de démontrer les diverses techniques efficaces pour l'évaluation de la vision chromatique humaine en utilisant des méthodes psychophysiques adaptatives.

El color es una cualidad perceptual que nos permite identificar y localizar estándares ambientales de mismo brillo y constituye una dimensión adicional en la identificación de los objetos, además de la detección de inúmeras otras cualidades de los objetos en su relación con la escena visual, como luminancia, contraste, forma, movimiento, textura, profundidad. Ahí está su importancia básica en las actividades hechas por los animales y por los seres humanos en su interacción con el ambiente. La psicofísica visual está preocupada del estudio cuantitativo de la relación entre los acontecimientos físicos de estimulación sensorial y la respuesta de comportamiento resultante de esta estimulación, suministrando de esta forma medios de evaluar aspectos de la visión humana, como la visión de colores. Este manuscrito tiene como objetivo demostrar diversas técnicas eficientes en la evaluación de la visión cromática humana a través de métodos psicofísicos adaptativos.