Dopamine and acetylcholine correlations in the nucleus accumbens depend on behavioral task states.

Dopamine in the nucleus accumbens ramps up as animals approach desired goals. These ramps have received intense scrutiny because they seem to violate long-held hypotheses on dopamine function. Furthermore, it has been proposed that they are driven by local acetylcholine release, i.e., that they are mechanistically separate from dopamine signals related to reward prediction errors. Here, we tested this hypothesis by simultaneously recording accumbal dopamine and acetylcholine signals in rats executing a task involving motivated approach. Contrary to recent reports, we found that dopamine ramps were not coincidental with changes in acetylcholine. Instead, we found that acetylcholine could be positively, negatively, or uncorrelated with dopamine depending on whether the task phase was determined by a salient cue, reward prediction error, or active approach, respectively. Our results suggest that accumbal dopamine and acetylcholine are largely independent but may combine to engage different postsynaptic mechanisms depending on the behavioral task states.

The role of the lateral orbitofrontal cortex in creating cognitive maps.

We use mental models of the world-cognitive maps-to guide behavior. The lateral orbitofrontal cortex (lOFC) is typically thought to support behavior by deploying these maps to simulate outcomes, but recent evidence suggests that it may instead support behavior by underlying map creation. We tested between these two alternatives using outcome-specific devaluation and a high-potency chemogenetic approach. Selectively inactivating lOFC principal neurons when male rats learned distinct cue-outcome associations, but before outcome devaluation, disrupted subsequent inference, confirming a role for the lOFC in creating new maps. However, lOFC inactivation surprisingly led to generalized devaluation, a result that is inconsistent with a complete mapping failure. Using a reinforcement learning framework, we show that this effect is best explained by a circumscribed deficit in credit assignment precision during map construction, suggesting that the lOFC has a selective role in defining the specificity of associations that comprise cognitive maps.

Neuroscience: Finding the middle ground of behavioral (in)flexibility.

We often perform actions even when they incur heavy costs or no longer serve any clear purpose. Two recent studies have independently identified heightened phasic dopamine release in the dorsomedial striatum as a predisposing factor for such behavioral inflexibility.

Dopamine.

Dopamine was first described by George Barger, James Ewens, and Henry Dale in 1910 as an epinephrine-like monoamine compound. Initially believed to be a mere precursor of norepinephrine, it was mostly ignored for the next four decades (Figure 1A). However, in the 1950s Kathleen Montagu showed that dopamine occurred in the brain by itself, and a series of studies by Arvid Carlsson and collaborators demonstrated that dopamine is a bona fide neurotransmitter, a finding that would earn Carlsson the 2000 Nobel Prize in Physiology and Medicine. In a landmark experiment, he pharmacologically blocked all dopamine neurotransmission in rabbits, which rendered them completely paralyzed, and then fully recovered their behavior with an injection of the dopamine precursor L-DOPA, demonstrating that dopamine was essential for self-initiated movement (Figure 1B). A similar effect was quickly reproduced by Oleg Hornykiewicz and collaborators in human Parkinsonian patients. Within a few years, dopamine jumped from relative obscurity to being critical for life as we know it.

Neuroscience: What, where, and how wonderful?

Hippocampal place cells represent spatial locations, but it is unclear how they incorporate associations between locations and specific outcomes. A recent study illuminates this issue by showing that place cells in intermediate hippocampus remap their fields following changes in reward.

The orbitofrontal cortex is necessary for learning to ignore.

Animals learn not only what is potentially useful but also what is meaningless and should be disregarded. How this is accomplished is a key but seldom explored question in psychology and neuroscience. Learning to ignore irrelevant cues is evident in latent inhibition-the ubiquitous phenomenon where presenting a cue several times without consequences leads to retardation of subsequent conditioning to that cue.1,2 Does learning to ignore these cues, because they predict nothing, involve the same neural circuits that are critical to learning to make predictions about other "real world" impending events? If so, the orbitofrontal cortex (OFC), as a key node in such networks, should be important.3 Specifically, the OFC has been hypothesized to participate in the recognition of hidden task states, which are not directly signaled by explicit outcomes.4 Evaluating its involvement in pre-exposure learning during latent inhibition would be an acid test for this hypothesis. Here, we report that selective chemogenetic inactivation of rat orbitofrontal cortex principal neurons during stimulus pre-exposure markedly reduces latent inhibition in subsequent conditioning. Inactivation only during pre-exposure ensured that the observed effects were due to an impact on the acquisition of information prior to its use in any sort of behavior, i.e., during latent learning. Further behavioral tests confirmed this, showing that the impact of OFC inactivation during pre-exposure was limited to the latent inhibition effect. These results demonstrate that the OFC is important for latent learning and the formation of associations even in the absence of explicit outcomes.

Sex-dependent alterations in behavior, drug responses and dopamine transporter expression in heterozygous DAT-Cre mice.

Heterozygous mice that express Cre-recombinase under the dopamine transporter promoter (DAT-Cre knock in mice, or KI) are widely used for targeting midbrain dopamine neurons, under the assumption that their constitutive physiology is not affected. We report here that these mice display striking sex-dependent behavioral and molecular differences in relation to wildtypes (WT). Male and female KI mice were constitutively hyperactive, and male KI mice showed attenuated hyperlocomotor responses to amphetamine. In contrast, female KIs displayed a marked reduction in locomotion ("calming" effect) in response to the same dose of amphetamine. Furthermore, male and female DAT-Cre KI mice showed opposing differences in reinforcement learning, with females showing faster conditioning and males showing slower extinction. Other behavioral variables, including working memory and novelty preference, were not changed compared to WT. These effects were paralleled by differences in striatal DAT expression that disproportionately affected female KI mice. Our findings reveal clear limitations of the DAT-Cre line that must be considered when using this model.

Dysfunctional suction dynamics in newborns with ankyloglossia / Dinâmica de sucção disfuncional em recém-nascidos com anquiloglossia

ABSTRACT Purpose Compare infant suction in babies with and without ankyloglossia using a microprocessor-controlled pressure sensor coupled to a pacifier. Methods Fifty-five infants from 0 to 2 months of age underwent clinical examination for ankyloglossia, after which they were offered a silicone pacifier connected to the pressure acquisition device and suction activity was recorded. Thus, we extracted the frequency of sucks within a burst, the average suck duration, the burst duration, the number of sucks per burst, the maximum amplitude of sucks per burst and the inter-burst interval. Results The key difference in newborns with ankyloglossia in relation to control was that they perform longer bursts of suction activity. Conclusion The longer burst durations are likely a compensatory strategy and may underlie the pain reported by mothers during breastfeeding. We therefore propose a method for objectively quantifying some parameters of infant suction capacity and demonstrate its use in assisting the evaluation of ankyloglossia.

RESUMO Objetivo Comparar a sucção infantil em bebês com e sem anquiloglossia usando um sensor de pressão controlado por microprocessador acoplado a uma chupeta. Método Cinquenta e cinco lactentes de 0 a 2 meses de idade foram submetidos ao exame clínico de anquiloglossia, em seguida foi oferecido uma chupeta de silicone conectada ao dispositivo de aquisição de pressão e a atividade de sucção foi registrada. Assim, obtivemos dados sobre a frequência de sucções dentro de um período de sucções, a duração média da sucção, a duração da rajada, o número de sucções por rajada, a amplitude máxima das sucções por rajada e o intervalo entre rajadas. O teste t não pareado foi utilizado para comparações entre os grupos. Resultados A principal diferença dos recém-nascidos com anquiloglossia em relação aos do grupo controle é que eles realizam rajadas mais longas durante a atividade de sucção. Conclusão A duração mais longa das rajadas é provavelmente uma estratégia compensatória e pode estar por trás da dor relatada pelas mães durante a amamentação. Portanto, propomos um método para quantificar objetivamente alguns parâmetros da sucção infantil e demonstramos seu uso para auxiliar na avaliação da anquiloglossia.

In vivo functional diversity of midbrain dopamine neurons within identified axonal projections.

Functional diversity of midbrain dopamine (DA) neurons ranges across multiple scales, from differences in intrinsic properties and connectivity to selective task engagement in behaving animals. Distinct in vitro biophysical features of DA neurons have been associated with different axonal projection targets. However, it is unknown how this translates to different firing patterns of projection-defined DA subpopulations in the intact brain. We combined retrograde tracing with single-unit recording and labelling in mouse brain to create an in vivo functional topography of the midbrain DA system. We identified differences in burst firing among DA neurons projecting to dorsolateral striatum. Bursting also differentiated DA neurons in the medial substantia nigra (SN) projecting either to dorsal or ventral striatum. We found differences in mean firing rates and pause durations among ventral tegmental area (VTA) DA neurons projecting to lateral or medial shell of nucleus accumbens. Our data establishes a high-resolution functional in vivo landscape of midbrain DA neurons.

Voice disorder in cystic fibrosis patients.

Cystic fibrosis is a common autosomal recessive disorder with drastic respiratory symptoms, including shortness of breath and chronic cough. While most of cystic fibrosis treatment is dedicated to mitigating the effects of respiratory dysfunction, the potential effects of this disease on vocal parameters have not been systematically studied. We hypothesized that cystic fibrosis patients, given their characteristic respiratory disorders, would also present dysphonic symptoms. Given that voice disorders can severely impair quality of life, the identification of a potential cystic fibrosis-related dysphonia could be of great value for the clinical evaluation and treatment of this disease. We tested our hypothesis by measuring vocal parameters, using both objective physical measures and the GRBAS subjective evaluation method, in male and female cystic fibrosis patients undergoing conventional treatment and compared them to age and sex matched controls. We found that cystic fibrosis patients had a significantly lower vocal intensity and harmonic to noise ratio, as well as increased levels of jitter and shimmer. In addition, cystic fibrosis patients also showed higher scores of roughness, breathiness and asthenia, as well as a significantly altered general grade of dysphonia. When we segregated the results according to sex, we observed that, as a group, only female cystic fibrosis patients had significantly lower values of harmonic to noise ratio and an abnormal general grade of dysphonia in relation to matched controls, suggesting that cystic fibrosis exerts a more pronounced effect on vocal parameters of women in relation to men. Overall, the dysphonic characteristics of CF patients can be explained by dysfunctions in vocal fold movement and partial upper airway obstruction, potentially caused by the accumulation of mucus and chronic cough characteristic of CF symptomatology. Our results show that CF patients exhibit significant dysphonia and suggest they may potentially benefit from voice therapy as a parallel treatment strategy.

Behavioral and cardiorespiratory responses to bilateral microinjections of oxytocin into the central nucleus of amygdala of Wistar rats, an experimental model of compulsion.

INTRODUCTION: The central nucleus of amygdala plays an important role mediating fear and anxiety responses. It is known that oxytocin microinjections into the central nucleus of amygdala induce hypergrooming, an experimental model of compulsive behavior. We evaluated the behavioral and cardiorespiratory responses of conscious rats microinjected with oxytocin into the central nucleus of amygdala. METHODS: Male Wistar rats were implanted with guide cannulae into the central nucleus of amygdala and microinjected with oxytocin (0.5 µg, 1 µg) or saline. After 24 h, rats had a catheter implanted into the femoral artery for pulsatile arterial pressure measurement. The pulsatile arterial pressure was recorded at baseline conditions and data used for cardiovascular variability and baroreflex sensitivity analysis. Respiratory and behavioral parameters were assessed during this data collection session. RESULTS: Microinjections of oxytocin (0.5 µg) into the central nucleus of amygdala produced hypergrooming behavior but did not change cardiorespiratory parameters. However, hypergrooming evoked by microinjections of oxytocin (1 µg) into the central nucleus of amygdala was accompanied by increase in arterial pressure, heart rate and ventilation and augmented the power of low and high (respiratory-related) frequency bands of the systolic arterial pressure spectrum. No changes were observed in power of the low and high frequency bands of the pulse interval spectrum. Baroreflex sensitivity was found lower after oxytocin microinjections, demonstrating that the oxytocin-induced pressor response may involve an inhibition of baroreflex pathways and a consequent facilitation of sympathetic outflow to the cardiovascular system. CONCLUSIONS: The microinjection of oxytocin (1 µg) into the central nucleus of amygdala not only induces hypergrooming but also changes cardiorespiratory parameters. Moreover, specific oxytocin receptor antagonism attenuated hypergrooming but did not affect pressor, tachycardic and ventilatory responses to oxytocin, suggesting the involvement of distinct neural pathways.

A new low cost wide-field illumination method for photooxidation of intracellular fluorescent markers.

Analyzing cell morphology is crucial in the fields of cell biology and neuroscience. One of the main methods for evaluating cell morphology is by using intracellular fluorescent markers, including various commercially available dyes and genetically encoded fluorescent proteins. These markers can be used as free radical sources in photooxidation reactions, which in the presence of diaminobenzidine (DAB) forms an opaque and electron-dense precipitate that remains localized within the cellular and organelle membranes. This method confers many methodological advantages for the investigator, including absence of photo-bleaching, high visual contrast and the possibility of correlating optical imaging with electron microscopy. However, current photooxidation techniques require the continuous use of fluorescent or confocal microscopes, which wastes valuable mercury lamp lifetime and limits the conversion process to a few cells at a time. We developed a low cost optical apparatus for performing photooxidation reactions and propose a new procedure that solves these methodological restrictions. Our "photooxidizer" consists of a high power light emitting diode (LED) associated with a custom aluminum and acrylic case and a microchip-controlled current source. We demonstrate the efficacy of our method by converting intracellular DiI in samples of developing rat neocortex and post-mortem human retina. DiI crystals were inserted in the tissue and allowed to diffuse for 20 days. The samples were then processed with the new photooxidation technique and analyzed under optical microscopy. The results show that our protocols can unveil the fine morphology of neurons in detail. Cellular structures such as axons, dendrites and spine-like appendages were well defined. In addition to its low cost, simplicity and reliability, our method precludes the use of microscope lamps for photooxidation and allows the processing of many labeled cells simultaneously in relatively large tissue samples with high efficacy.

Scopolamine- and diazepam-induced amnesia are blocked by systemic and intraseptal administration of substance P and choline chloride.

Systemic (IP) and/or intraseptal (IS) administration of scopolamine (SCP) and diazepam (DZP) induce amnesia, whereas IP injection of the neuropeptide substance P (SP) and choline chloride (ChCl) produce memory facilitation. The septohippocampal cholinergic system has been pointed out as a possible site of SCP and DZP-induced amnesia as well as for the mnemonic effects induced by SP and ChCl. We performed a series of experiments in order to investigate the interactions between cholinergic and GABA/benzodiazepine (GABA/BZD) systems with the SPergic system on inhibitory avoidance retention. Male Wistar rats were trained and tested in a step-down inhibitory avoidance task (1.0 mA footshock). Animals received, pre-training, IP (1.0 mg/kg) or IS (1.0 nM/0.5 microl) injection of DZP, SCP (SCP; 1.0 mg/kg - IP or 0.5 microM/0.5 microl--IS) or vehicle (VEH). Immediately after training they received an IP or IS injections of SP 1-11 (50 microg/kg--IP or 1.0 nM/0.5 microl--IS), SP 1-7 (167 microg/kg--IP or 1.0 nM/0.5 microl--IS), ChCl (20 mg/kg--IP or 0.3 microM/0.5 microl--IS) or VEH. Rats pretreated with SCP and DZP showed amnesia. Post-trial treatments with SP 1-11, SP 1-7 or ChCl blocked the amnesic effects of SCP and DZP. These findings suggest an interaction between SPergic and cholinergic mechanisms with GABAergic systems in the modulation of inhibitory avoidance retention and that the effects of these treatments are mediated, at least in part, by interactions in the septohippocampal pathway.

Respostas comportamentais do camarão de água doce Macrobrachium amazonicum à estimulação por cloreto de amônia / Behavioural responses of the freshwater prawn Macrobrachium amazonicum to chemical with ammonium chloride

O objetivo do presente estudo foi analisar as respostas comportamentais do camarão de água doce Macrobrachium amazonicum, mediante estimulação química com cloreto de amônia a 100mM. Espécimes desse crustáceo foram posicionados em um aquário com quatro litros de água doce desclorificada, e um pulso de solução estimulante foi injetado no recipiente na extremidade oposta em relação à posição do animal. Os animais estudados não apresentaram resposta aos estímulos utilizados. Isso pode sugerir que o limiar de resposta comportamental é maior do que o limiar fisiológico dos neurônios receptivos primários, ou que o cloreto de amônia precisa ser associado com outros compostos ou determinadas variáveis físicas para poder gerar uma resposta ecológica na espécie estudada.

The objective of the present study was to analyze the behavioral responses of the freshwater prawn Macrobrachium amazonicum, elicited by stimulation with ammonium chloride at 100mM. Specimens were positioned in a four liter dechlorinated freshwater tank and a 0.5ml pulse of stimulant was applied in the opposing corner of the recipient. The studied animals demonstrated no response to the stimuli applied. This may suggest that the behavioral response threshold is higher than the physiological response threshold of the primary chemoreceptor neurons or that the ammonium chloride must be associated with other compounds or certain physical attributes in order to evoke an ethological reaction from the studied species.

Respostas comportamentais do camarão de água doce Macrobrachium amazonicum à estimulação por cloreto de amônia / Behavioural responses of the freshwater prawn Macrobrachium amazonicum to chemical with ammonium chloride

O objetivo do presente estudo foi analisar as respostas comportamentais do camarão de água doce Macrobrachium amazonicum, mediante estimulação química com cloreto de amônia a 100mM. Espécimes desse crustáceo foram posicionados em um aquário com quatro litros de água doce desclorificada, e um pulso de solução estimulante foi injetado no recipiente na extremidade oposta em relação à posição do animal. Os animais estudados não apresentaram resposta aos estímulos utilizados. Isso pode sugerir que o limiar de resposta comportamental é maior do que o limiar fisiológico dos neurônios receptivos primários, ou que o cloreto de amônia precisa ser associado com outros compostos ou determinadas variáveis físicas para poder gerar uma resposta ecológica na espécie estudada.(AU)

The objective of the present study was to analyze the behavioral responses of the freshwater prawn Macrobrachium amazonicum, elicited by stimulation with ammonium chloride at 100mM. Specimens were positioned in a four liter dechlorinated freshwater tank and a 0.5ml pulse of stimulant was applied in the opposing corner of the recipient. The studied animals demonstrated no response to the stimuli applied. This may suggest that the behavioral response threshold is higher than the physiological response threshold of the primary chemoreceptor neurons or that the ammonium chloride must be associated with other compounds or certain physical attributes in order to evoke an ethological reaction from the studied species.(AU)