Telencephalic distributions of doublecortin and glial fibrillary acidic protein suggest novel migratory pathways in adult lizards.

Adult neurogenesis has been reported in all major vertebrate taxa. However, neurogenic rates and the number of neurogenic foci vary greatly, and are higher in ancestral taxa. Our study aimed to evaluate the distribution of doublecortin (DCX) and glial fibrillary acidic protein (GFAP) in telencephalic areas of the adult tropical lizard Tropidurus hispidus. We describe evidence for four main neurogenic foci, which coincide anatomically with the ventricular sulci described by the literature. Based on neuronal morphology, we infer four migratory patterns/pathways. In the cortex, patterns of GFAP and DCX staining support radial migrations from ventricular zones into cortical areas and dorsoventricular ridge. Cells radiating from the sulcus septomedialis (SM) seemed to migrate to the medial cortex and dorsal cortex. From the sulcus lateralis (SL), they seemed to be bound for the lateral cortex, central amygdala and nucleus sphericus. We describe a DCX-positive stream originating in the caudal sulcus ventralis and seemingly bound for the olfactory bulb, resembling a rostral migratory stream. We provide evidence for a previously undescribed tangential dorso-septo-caudal migratory stream, with neuroblasts supported by DCX-positive fibers. Finally, we provide evidence for a commissural migration stream seemingly bound for the contralateral nucleus sphericus. Therefore, in addition to two previously known migratory streams, this study provides anatomical evidence in support for two novel migratory routes in amniotes.

Expression of aggressiveness modulates mesencephalic c-fos activation during a social interaction test in Japanese quail (Coturnix japonica).

It is well known that during a social conflict, interactions are dependent on the animal's propensity to behave aggressively as well as the behavior of the opponent. However, discriminating between these two confounding factors was difficult. Recently, a Social Interaction (SI) test using photocastrated males as non-aggressive stimuli was proposed as a useful tool to evaluate aggressiveness. The avian Intercollicular- Griseum centralis complex (comparable to mammalian periaqueductal gray) has been reported as a crucial node in the descending pathways that organize behavioral and autonomic aspects of defensive responses and aggressiveness. Herein, using the SI test, we evaluated whether mesencephalic areas are activated (expressed c-fos) when photostimulated adult males are confronted with non-responsive (non-aggressive) opponents. Furthermore, we also examined whether mesencephalic activation is related to male performance during the SI test (i.e., aggressive vs. non-aggressive males) in birds reared in enriched or in standard environments. Five mesencephalic areas at two anatomic levels (intermediate and rostral) and locomotion during SI testing were studied. Aggressive males showed increased c-fos expression in all areas studied, and moved at faster speeds in comparison to their non-aggressive and control counterparts. Non-aggressive males and the test controls showed similar c-fos labeling. In general, rearing condition did not appear to influence c-fos expression nor behavior during the SI test. Findings suggest that mesencephalic activation is involved when males are actively expressing aggressive behaviors. This overall phenomenon is shown regardless of both the environmental stimuli provided during the birds´ rearing and the potentially stressful stimuli during the SI trial.

Distribution of serotonin 5-HT1A-binding sites in the brainstem and the hypothalamus, and their roles in 5-HT-induced sleep and ingestive behaviors in rock pigeons (Columba livia).

Serotonin 1A receptors (5-HT1ARs), which are widely distributed in the mammalian brain, participate in cognitive and emotional functions. In birds, 5-HT1ARs are expressed in prosencephalic areas involved in visual and cognitive functions. Diverse evidence supports 5-HT1AR-mediated 5-HT-induced ingestive and sleep behaviors in birds. Here, we describe the distribution of 5-HT1ARs in the hypothalamus and brainstem of birds, analyze their potential roles in sleep and ingestive behaviors, and attempt to determine the involvement of auto-/hetero-5-HT1ARs in these behaviors. In 6 pigeons, the anatomical distribution of [(3)H]8-OH-DPAT binding in the rostral brainstem and hypothalamus was examined. Ingestive/sleep behaviors were recorded (1h) in 16 pigeons pretreated with MM77 (a heterosynaptic 5-HT1AR antagonist; 23 or 69 nmol) for 20 min, followed by intracerebroventricular ICV injection of 5-HT (N:8; 150 nmol), 8-OH-DPAT (DPAT, a 5-HT1A,7R agonist, 30 nmol N:8) or vehicle. 5-HT- and DPAT-induced sleep and ingestive behaviors, brainstem 5-HT neuronal density and brain 5-HT content were examined in 12 pigeons, pretreated by ICV with the 5-HT neurotoxin 5,7-dihydroxytryptamine (5,7-DHT) or vehicle (N:6/group). The distribution of brainstem and diencephalic c-Fos immunoreactivity after ICV injection of 5-HT, DPAT or vehicle (N:5/group) into birds provided with or denied access to water is also described. 5-HT1ARs are concentrated in the brainstem 5-HTergic areas and throughout the periventricular hypothalamus, preoptic nuclei and circumventricular organs. 5-HT and DPAT produced a complex c-Fos expression pattern in the 5-HT1AR-enriched preoptic hypothalamus and the circumventricular organs, which are related to drinking and sleep regulation, but modestly affected c-Fos expression in 5-HTergic neurons. The 5-HT-induced ingestivebehaviors and the 5-HT- and DPAT-induced sleep behaviors were reduced by MM77 pretreatment. 5,7-DHT increased sleep per se, decreased tryptophan hydroxylase expression in the raphe nuclei and decreased prosencephalic 5-HT release but failed to affect 5-HT- or DPAT-induced drinking or sleep behavior. 5-HT- and DPAT-induced ingestive and sleep behaviors in pigeons appear to be mediated by heterosynaptic and/or non-somatodendritic presynaptic 5-HT1ARs localized to periventricular diencephalic circuits.

Stress-induced core temperature changes in pigeons (Columba livia).

Changes in body temperature are significant physiological consequences of stressful stimuli in mammals and birds. Pigeons (Columba livia) prosper in (potentially) stressful urban environments and are common subjects in neurobehavioral studies; however, the thermal responses to stress stimuli by pigeons are poorly known. Here, we describe acute changes in the telemetrically recorded celomatic (core) temperature (Tc) in pigeons given a variety of potentially stressful stimuli, including transfer to a novel cage (ExC) leading to visual isolation from conspecifics, the presence of the experimenter (ExpR), gentle handling (H), sham intracelomatic injections (SI), and the induction of the tonic immobility (TI) response. Transfer to the ExC cage provoked short-lived hyperthermia (10-20 min) followed by a long-lasting and substantial decrease in Tc, which returned to baseline levels 2 h after the start of the test. After a 2-hour stay in the ExC, the other potentially stressful stimuli evoked only weak, marginally significant hyperthermic (ExpR, IT) or hypothermic (SI) responses. Stimuli delivered 26 h after transfer to the ExC induced definite and intense increases in Tc (ExpR, H) or hypothermic responses (SI). These Tc changes appear to be unrelated to modifications in general activity (as measured via telemetrically recorded actimetric data). Repeated testing failed to affect the hypothermic responses to the transference to the ExC, even after nine trials and at 1- or 8-day intervals, suggesting that the social (visual) isolation from conspecifics may be a strong and poorly controllable stimulus in this species. The present data indicated that stress-induced changes in Tc may be a consistent and reliable physiological parameter of stress but that they may also show stressor type-, direction- and species-specific attributes.

Spontaneous absence-like activity in Wistar rats: Behavioral and electrographic characteristics and the effects of antiepileptic drugs / Atividade epiléptica espontânea tipo ausência em Ratos Wistar: características eletrográficas e comportamentais e os efeitos de drogas antiepilépticas

Current investigation describes the behavioral and electrographic characteristics of spontaneous absence-like seizures identified in Wistar rats (referred to here as FMUSP-rats, after the Faculty of Medicine, University of São Paulo, São Paulo State, Brazil), and characterized by spike -wave discharges (SWDs) in the neocortex and the hippocampus. After consanguineous crossing directed to an increased incidence of seizures, the latter were observed in almost all F9 offspring. FMUSP-rat seizures are expressed as immobility and concomitant SWDs, oscillating between 7.5 and 12 Hz in the frontoparietal cortex and the hippocampus. Behaviorally, they are mainly associated with clonic movements of the eyes, rostrum and vibrissae, the latter ranging between 1 and 70 seconds and occur at a rate of up to 229 per hour. Systemic injections of ethosuximide (0, 25, 50, 100, 250 mg kg-1) and of diazepam (15 mg kg-1) increased the latency for the first seizure and reduced both the hourly incidence of SWD bursts and their mean duration. Carbamazepine (30 mg kg-1) injections increased both the incidence and duration of the SWDs, leaving the latency for the first seizure unchanged. Comparisons between FMUSP-rats and well-established genetic models of absence seizures data indicated that the animals described herein might contribute towards studies on the neurological condition under analysis.

Neste estudo, descrevemos aspectos eletrográficos e comportamentais de atividade semelhante às crises de ausência identificadas em ratos Wistar (ratos FMUSP - Faculdade de Medicina da Universidade de São Paulo, Estado de São Paulo, Brasil), que são caracterizadas por descargas em forma de espícula-onda no neocórtex e no hipocampo em 100% dos ratos da geração F9 resultante de cruzamentos consanguíneos. As crises se manifestaram com imobilidade comportamental associada com atividade eletrográfica em forma de espícula -onda oscilando entre 7,5 e 12 Hz no córtex frontoparietal e no hipocampo. Também foram observados comportamentos associados como clonias oculares, rostrais e de vibrissa que duravam de 1 a 70 segundos podendo ocorrer a uma taxa de até 229 eventos por hora. Injeçõe sistêmicas de etosuximida (0, 25, 50, 100 e 250 mg kg-1) and of diazepam (15 mg kg-1) aumentaram a latência para a primeira crise e reduziram tanto a incidência quanto a duração das crises. Injeções de Carbamazepina (30 mg kg-1) aumentaram tanto a incidência quanto a duração das crises sem interferir na latência. Comparações feitas entre os ratos epilépticos FMUSP com os modelos genéticos de epilepsia bem estabelecidos na atualidade indicaram que o modelo aqui descrito pode contribuir para um melhor entendimento dos mecanismos relacionados a esta condição neurológica.

The behavioral satiety sequence in pigeons (Columba livia). Description and development of a method for quantitative analysis.

The postprandial event known as the specific dynamic action is an evolutionarily conserved physiological set of metabolic responses to feeding. Its behavioral counterpart, a sequence of drinking, maintenance (e.g., grooming) and sleep-like behaviors known as the behavioral satiety sequence (BSS), has been thoroughly described in rodents and has enabled the refined evaluation of potential appetite modifiers. However, the presence and attributes of a BSS have not been systematically studied in non-mammalian species. Here, we describe the BSS induced in pigeons (Columba livia) by 1) the presentation of a palatable seed mixture (SM) food to free-feeding animals (SM+FF condition) and 2) re-feeding after a 24-h fasting period (FD24h+SM), which was examined by continuous behavioral recording for 2h. We then compare these patterns to those observed in free-feeding (FF) animals. A set of graphic representations and indexes, drawn from these behaviors (latency, time-to-peak, inter-peak intervals and the first intersection between feeding curves and those of other BSS-typical behaviors) were used to describe the temporal structure and sequential relationships between the pigeon's BSS components. Cramér-von Mises-based statistical procedures and bootstrapping-based methods to compare pairs of complex behavioral curves were described and used for comparisons among the behavioral profiles during the free-feeding recordings and after fasting- and SM-induced BSS. FD24h+SM- and SM+FF-induced feeding were consistently followed by a similar sequence of increased bouts of drinking, followed by preening and then sleep, which were significantly different from that of FF birds. The sequential and temporal patterns of the pigeon's BSS were not affected by differences in food intake or by dissimilarity in motivational content of feeding stimuli. The present data indicated that a BSS pattern can be reliably evoked in the pigeon, in a chronological succession and sequence that strongly resembled that observed in rodents and primates. This pattern can be quantitatively described and compared using different suitable and coordinated behavioral measures, enabling further studies on the comparative and evolutionary aspects of the mechanisms that shape the post-consummatory behavioral flux in amniotes.

Serotonergic control of ingestive and post-ingestive behaviors in pigeons (Columba livia): the role of 5-HT1A receptor-mediated central mechanisms.

Central injections of serotonin (5-HT) produce hyperdipsic and hypnogenic behavioral effects that are correlated to decreased Fos-immunorreactivity of 5-HT neurons in free-feeding pigeons. We herein (1) probed the role of 5-HT(1A) receptors on the 5-HT- or 8-OH-DPAT-evoked postprandial behaviors and (2) described the sleep-waking states (waking, W; drowsiness, D; slow-wave sleep, SWS; rapid-eye movement sleep, REMS) and sleep architecture of free-feeding pigeons after these treatments. Latency, frequency and duration of feeding, drinking, preening, exploratory and sleep-like behaviors (SLB) were examined after intracerebroventricular (ICV) injections of 5-HT (0, 50 or 150 nmol) or 8-OH-DPAT (DPAT, 0 or 30 nmol) in pigeons pretreated with the 5-HT(1A) antagonist WAY100635 (WAY, 0, 0.1, 0.3 or 1 nmol). Additionally, the acute (1h) waking-sleep-related electrographic activity in the hippocampus (HP) was examined after ICV injections of 5-HT (150 or 300 nmol) or DPAT (30 or 60 nmol) in pigeons pretreated with WAY (0 or 1 nmol). 5-HT and DPAT acutely increased drinking and then sleep: all doses of WAY attenuated the 5-HT (50 nmol) -induced dipsogenic effect, but left unchanged the effects of the 150 nmol 5-HT dose. The WAY 0.1 nmol dose blocked the SLB induced by the 5-HT 50 nmol dose. Given before the vehicle (VEH) injections, WAY does not affect water or food intake, but increased the SLB duration at all doses. DPAT injections increased feeding, drinking and SLB. All the WAY doses attenuated the DPAT-induced drinking and feeding responses, and the WAY 0.1 and 0.3 nmol doses reduced DPAT-induced SLB. DPAT or 5-HT injections decreased the duration of electrographically-determined waking, increased the durations of D and induced the emergence of SWS and REMS states indistinguishable from the hippocampal EEG associated with spontaneous sleep, as judged from visual and spectral analysis. WAY (1 nmol) increased SWS and D, and potentiated the 5-HT- and DPAT-induced SWS. These data suggest that 5-HT-induced drinking depends on the activation of presynaptic 5-HT(1A) receptors, while 5-HT(1A) autoreceptor activation contributes to the 5-HT-induced sleep. 5-HT-induced drinking and sleep behaviors may thus be provoked by a 5-HT(1A)-evoked, rebound-like reduction in central 5-HTergic activity. These data also indicate that an ongoing, tonic and inhibitory influence of central 5-HT circuits may participate in the control of feeding, drinking and rest behaviors in pigeons during the wake, nibbling diurnal state. These mechanisms appear to be comparable to those found in mammals, suggesting that they may represent a conserved, plesiomorphic functional trait of the amniotes brain.

NMDA preconditioning attenuates cortical and hippocampal seizures induced by intracerebroventricular quinolinic acid infusion.

Searching for new therapeutic strategies through modulation of glutamatergic transmission using effective neuroprotective agents is essential. Glutamatergic excitotoxicity is a common factor to neurodegenerative diseases and acute events such as cerebral ischemia, traumatic brain injury, and epilepsy. This study aimed to evaluate behavioral and electroencephalographic (EEG) responses of mice cerebral cortex and hippocampus to subconvulsant and convulsant application of NMDA and quinolinic acid (QA), respectively. Moreover, it aimed to evaluate if EEG responses may be related to the neuroprotective effects of NMDA. Mice were preconditioned with NMDA (75 mg/kg, i.p.) and EEG recordings were performed for 30 min. One day later, QA was injected (36.8 nmol/site) and EEG recordings were performed during 10 min. EEG analysis demonstrated NMDA preconditioning promotes spike-wave discharges (SWDs), but it does not display behavioral manifestation of seizures. Animals that were protected by NMDA preconditioning against QA-induced behavioral seizures, presented higher number of SWD after NMDA administration, in comparison to animals preconditioned with NMDA that did display behavioral seizures after QA infusion. No differences were observed in latency for the first seizure or duration of seizures. EEG recordings after QA infusion demonstrated there were no differences in the number of SWD, latency for the first seizure or duration of seizures in animals pretreated with saline or in animals preconditioned by NMDA that received QA. A negative correlation was identified between the number of NMDA-induced SWD and QA-induced seizures severity. These results suggest a higher activation during NMDA preconditioning diminishes mice probability to display behavioral seizures after QA infusion.

Feeding behaviour after injection of α-adrenergic receptor agonists into the median raphe nucleus of food-deprived rats.

This study investigated the participation of median raphe nucleus (MnR) α1-adrenergic receptors in the control of feeding behaviour. The α1-adrenergic agonist phenylephrine (PHE) and α2-adrenergic agonist clonidine (CLON) (at equimolar doses of 0, 6 and 20 nmol) were injected into the MnR of: a) rats submitted to overnight fasting (18 h); or b) rats maintained with 15 g of lab chow/day for 7 days. Immediately after the drug injections, the animals were placed in the feeding chamber and feeding and non-ingestive behaviours such as grooming, rearing, resting, sniffing and locomotion were recorded for 30 min. The results showed that both doses of PHE injected into the MnR of overnight fasted animals decreased food intake accompanied by an increase in the latency to start feeding. A reduction in feeding duration was observed only after treatment of the MnR with the 20 nmol dose of PHE. Both locomotion duration and sniffing frequency increased after injection with the highest dose PHE into the MnR. Feeding frequency and the other non-ingestive behaviours remained unchanged after PHE treatment in the MnR. Both doses of PHE injected into the MnR of food-restricted rats decreased food intake. This hypophagic response was accompanied by a decrease in feeding duration only after treatment of the MnR with the highest dose of PHE. The latency to start feeding and feeding frequency were not affected by injection of either dose of PHE into the MnR. While both doses of PHE increased sniffing duration, the highest dose of PHE increased resting duration and resting frequency. Treatment with CLON into the MnR did not affect feeding behaviour in either of the food deprivation conditions. The present results indicate the inhibitory functional role of α1-adrenergic receptors within the MnR on feeding behaviour.

Effects of GABA ligands injected into the nucleus accumbens shell on fear/anxiety-like and feeding behaviours in food-deprived rats.

In an attempt to establish a relationship between food intake and fear/anxiety-related behaviours, the goal of this study was to investigate the effect of bilateral injections of GABAA (Muscimol, MUS, doses 25 and 50ng/side) and GABAB (Baclofen, BAC, doses 32 and 64ng/side) receptor agonists in the nucleus accumbens shell (AcbSh) on the level of fear/anxiety-like and feeding behaviours in 24h food-deprived rats. The antagonists of GABAA (Bicuculline, BIC, doses 75 and 150ng/side) and GABAB (Saclofen, SAC, doses 1.5 and 3µg/side) were also tested. The results indicated that the total number of risk assessment behaviour decreased after the injection of both doses of GABAA agonist (MUS) into the AcbSh of 24h food-deprived rats exposed to elevated plus maze. Similar results were obtained after treatment with both doses of GABAB (BAC) agonist in the AcbSh. These data indicated that the activation of both GABAA and GABAB receptors within the AcbSh caused anxiolysis in 24h food-deprived rats. In addition, feeding behaviour (food intake, feeding latency and feeding duration) remained unchanged after treatment with both GABA agonists. In contrast, both food intake and feeding duration decreased after injections of both doses of BIC (GABAA antagonist), while the feeding latency remained unchanged after treatment with both GABA antagonists in the AcbSh of 24h food-deprived rats. The treatment with SAC (GABAB antagonist) did not affect feeding behaviour. Collectively, these data suggest that emotional changes evoked by pharmacological manipulation of the GABA neurotransmission in the AcbSh are not linked with changes in food intake.

ETHOWATCHER: validation of a tool for behavioral and video-tracking analysis in laboratory animals.

We present a software (ETHOWATCHER(®)) developed to support ethography, object tracking and extraction of kinematic variables from digital video files of laboratory animals. The tracking module allows controlled segmentation of the target from the background, extracting image attributes used to calculate the distance traveled, orientation, length, area and a path graph of the experimental animal. The ethography module allows recording of catalog-based behaviors from environment or from video files continuously or frame-by-frame. The output reports duration, frequency and latency of each behavior and the sequence of events in a time-segmented format, set by the user. Validation tests were conducted on kinematic measurements and on the detection of known behavioral effects of drugs. This software is freely available at www.ethowatcher.ufsc.br.

Alpha1 receptor antagonist in the median raphe nucleus evoked hyperphagia in free-feeding rats.

Serotonergic neurons in the median raphe nucleus (MnR) are stimulated by α(1)-adrenergic agonists and inhibited by α(2)-agonists. This study investigated the effect of the blockade of the MnR α(1)-adrenergic receptors of free feeding rats as an attempt to elucidate the functional role of these receptors in the control of feeding behavior. In addition, an α(2)-receptor antagonist was also administered in the MnR in order to strengthen the previous suggestion that α(2)-adrenergic receptors participate in the control of feeding behavior, probably decreasing the facilitatory influence on MnR serotonergic neurons. The α(1)-adrenergic antagonist prazosin (PRA, 40 nmol) or vehicle was injected into the MnR 15 min before treatment with phenylephrine (PHE, 0.2 nmol). The α(2)-adrenergic antagonist yohimbine (YOH, 40 nmol) was administered 15 min before clonidine (CLO, 20 nmol) or vehicle in free-feeding rats. After the injections, the animals were placed in the feeding chamber for 30 min to evaluate the ingestive and non-ingestive behaviors. At the end of the experiment the quantity of food and water consumed were measured. While treatment with PRA in the MnR followed by PHE did not change the feeding behavior, PRA injection alone into the MnR caused hyperphagia accompanied by a reduction in the latency to start eating, an increase in feeding frequency and an increase in the feeding duration. Pretreatment with YOH in the MnR blocked the hyperphagic effect induced by CLO. The present data reinforce our previous suggestion that the MnR α(2)-adrenergic receptors participate in the control of feeding behavior, probably decreasing the facilitatory influence on MnR serotonergic neurons of free-feeding animals. Furthermore, these results indicate that this influence is tonically mediated by α(1)-adrenergic receptors upon MnR neurons, which inhibit food intake.

Behavioral profile and Fos activation of serotonergic and non-serotonergic raphe neurons after central injections of serotonin in the pigeon (Columba livia).

Central injections of serotonin (5-HT) in food-deprived/refed pigeons evoke a sequence of hypophagic, hyperdipsic and sleep-like responses that resemble the postprandial behavioral sequence. Fasting-refeeding procedures affect sleep and drinking behaviors "per se". Here, we describe the behavioral profile and long-term food/water intake following intracerebroventricular (ICV) injections of 5-HT (50, 150, 300 nmol/2 µl) in free-feeding/drinking pigeons. The patterns of Fos activity (Fos+) in serotonergic (immunoreactive to tryptophan hydroxylase, TPH+) neurons after these treatments were also examined. 5-HT ICV injections evoked vehement drinking within 15 min, followed by an intense sleep. These effects did not extend beyond the first hour after treatment. 5-HT failed to affect feeding behavior consistently. The density of double-stained (Fos+/TPH+) cells was examined in 6 brainstem areas of pigeons treated with 5-HT (5-HTW) or vehicle. Another group received 5-HT and remained without access to water during 2h after treatment (5-HTØ). In the pontine raphe, Fos+ density correlated positively to sleep, and increased in both the 5-HTW and 5-HTØ animals. In the n. linearis caudalis, Fos+ and Fos+/TPH+ labeling was negatively correlated to sleep and was reduced in 5-HTØ animals. In the A8 region, Fos+/TPH+ labeling was reduced in 5-HTW and 5-HTØ animals, was positively correlated to food intake and negatively correlated to sleep. These data indicate that hyperdipsic and hypnogenic effects of ICV 5-HT in pigeons may result from the inhibition of a tonic activity of serotonergic neurons, which is possibly relevant to the control of postprandial behaviors, and that these relationships are shared functional traits of the serotonergic circuits in amniotes.

Assessment of observers' stability and reliability - a tool for evaluation of intra- and inter-concordance in animal behavioral recordings.

Behavior studies on the neurobiological effects of environmental, pharmacological and physiological manipulations in lab animals try to correlate these procedures with specific changes in animal behavior. Parameters such as duration, latency and frequency are assessed from the visually recorded sequences of behaviors, to distinguish changes due to manipulation. Since behavioral recording procedure is intrinsically interpretative, high variability in experimental results is expected and usual, due to observer-related influences such as experience, knowledge, stress, fatigue and personal biases. Here, we present a computer program that supports the assessment of inter- and intra-observer concordance, using statistical indices (e.g., Kappa and Kendal coefficients and concordance index). The software was tested in a case study with 4 different observers, naïve to behavioral recording procedures. On paired analysis, the higher agreement index achieved was 0.76 (concordance index) and 0.47 (Kappa Coefficient, where 0 is no agreement and 1 is total agreement). Observers showed poor concordance indices (lower than 0.7), emphasizing the concern on observer recording stability and on precise morphological definition of the recorded behaviors. These indices can also be used to train observers and to refine the behavioral catalogue definitions, as they are related to different behavioral recording aspects.

Proanthocyanidin-rich fraction from Croton celtidifolius Baill confers neuroprotection in the intranasal 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine rat model of Parkinson's disease.

We have recently demonstrated that rodents treated intranasally with 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) suffered impairments in olfactory, cognitive and motor functions associated with time-dependent disruption of dopaminergic neurotransmission in different brain structures conceivably analogous to those observed during different stages of Parkinson's disease (PD). On the other hand, the proanthocyanidin-rich fraction (PRF) obtained from the bark of Croton celtidifolius Baill (Euphorbiaceae), a tree frequently found in the Atlantic forest in south Brazil, has been described to have several neurobiological activities including antioxidant and anti-inflammatory properties, which may be of interest in the treatment of PD. The present data indicated that the pretreatment with PRF (10 mg/kg, i.p.) during five consecutive days was able to prevent mitochondrial complex-I inhibition in the striatum and olfactory bulb, as well as a decrease of the enzyme tyrosine hydroxylase expression in the olfactory bulb and substantia nigra of rats infused with a single intranasal administration of MPTP (1 mg/nostril). Moreover, pretreatment with PRF was found to attenuate the short-term social memory deficits, depressive-like behavior and reduction of locomotor activity observed at different periods after intranasal MPTP administration in rats. Altogether, the present findings provide strong evidence that PRF from C. celtidifolius may represent a promising therapeutic tool in PD, thus being able to prevent both motor and non-motor early symptoms of PD, together with its neuroprotective potential.

WAY100635 blocks the hypophagia induced by 8-OH-DPAT in the hypothalamic nuclei.

This study examined the influence of pretreatment with N-[2-[4-(2-methoxyphenyl)-1-piperazinyl]ethyl]-N-(2-pyridinyl) cyclohexane carboxamide maleate (WAY100635, full 5-HT1A receptor antagonist, 37 nmol) on feeding effects evoked by local injections of 8-hydroxy-2-(di-n-propylamino)-tetralin (8-OH-DPAT, 5-HT(1A) and 5-HT(7) receptor agonist, 6 nmol) into the LH and into the ARC of female rats adapted to a wet mash diet (enriched with 10% sucrose), during diestrus or estrus. The results showed that the LH-pretreatment with WAY100635 suppressed the hypophagic effects evoked by 8-OH-DPAT during estrus as well as diestrus. The ARC pretreatment with WAY100635 blockaded the hypophagia evoked by 8-OH-DPAT in estrus rats. The previous treatment with WAY100635 in the ARC also suppressed the feeding duration decrease evoked by 8-OH-DPAT in estrus. The latency to start feeding, the drinking behavior and the durations of other non-ingestive behaviors were not affected by the different treatments, hypothalamic regions (LH or ARC), and/or estrous cycle stages (diestrus and estrus), except for the locomotion duration increase after 8-OH-DAPT in LH-pretreated rats in diestrus. The present findings confirm our previous suggestion that ARC- and the LH-5-HT(1A) receptors participate in the serotonergic control of feeding and that these feeding-related serotonergic circuits in LH are affected by ovarian hormones, since the treatment with WAY100635 evoked a hypophagia response during the diestrus phases.

Arcopallium, NMDA antagonists and ingestive behaviors in pigeons.

This study investigated the effects of local injections of 6,7-dinitroquinoxaline-2,3(1H,4H)-dione (DNQX, AMPA-kainate receptor antagonist, 0.8 and 2.7 nmol) and MK-801 (NMDA receptor antagonist, 1.8 and 6.0 nmol) into the nucleus taeniae of the amygdala (TnA) and the arcopallium intermedium (AI) on ingestive and non-ingestive behaviors in free-feeding pigeons. Injections of DNQX into the TnA or into the AI failed to consistently affect feeding behavior; DNQX vehicle (DMSO) affected drinking when injected into the TnA. MK-801 injections into the AI produced a delayed increase in food and water intake. In the TnA, MK-801 increased water intake in the first two hours after the treatment. These data indicate that glutamatergic circuits in arcopallial structures in the pigeon, comparable to the mammalian medial amygdala, are involved in the inhibitory control of ingestive behaviors, suggesting that this can represent a conserved functional attribute in the amniote prosencephalon.

Tonic serotonergic control of ingestive behaviours in the pigeon (Columba livia): the role of the arcopallium.

This study examined the acute changes in feeding and drinking behaviours of free-feeding and free-drinking pigeons, in response to local injections of metergoline (MET, 5-HT(1/2) receptor antagonist; 7 and 20 nmol), GR46611 (GR, 5-HT(1B/1D) agonist; 2 and 6 nmol) or vehicle, into two components of the arcopallium: the nucleus taeniae of the amygdala (TnA) and the arcopallium intermedium (AI). In the TnA, the highest MET dose elicited a short-lived hyperphagy, without affecting drinking or non-ingestive behaviours during the first hour after injection. In contrast, all MET doses promptly increased drinking when injected in the AI, without affecting feeding; this effect was still evident 3 and 24 h after the treatment. When injected in the TnA, the highest GR dose promptly increased both food and water intake; these effects persisted 24 h after the treatments. GR injections in the AI evoked long-lasting increases in drinking, but not in feeding. Injections of these drugs into other arcopallial nuclei evoked no significant ingestive effects. These data indicate the presence of a tonic inhibitory influence of serotonergic inputs, partially mediated by 5-HT(1B/1D) receptors, on feeding- and drinking-related TnA circuits and on mechanisms controlling drinking in the AI. Compared to data from the rodent medial amygdala, our results suggest that a tonic inhibitory 5-HTergic control of feeding (but not drinking) behaviour, mediated by 5-HT(1/2) receptors and exerted in the medial amygdaloid area, may represent a broadly conserved functional attribute in the amniote brain, but probably involves many important taxa-specific neural mechanisms.

Distribution of tryptophan hydroxylase-immunoreactive neurons in the brainstem and diencephalon of the pigeon (Columba livia).

The distribution of tryptophan hydroxylase (TPH)-containing perikarya and processes in the brainstem and diencephalon of the pigeon (Columba livia) were investigated using single-labeling chromogenic and double-labeling fluorescence immunohistochemical methods for TPH and 5-HT. TPH-immunoreactive (TPH-ir) perikarya were seen extending from the caudal medulla to mid-hypothalamic levels, located in brainstem regions previously described as containing 5-HT-ir somata. Brainstem TPH-ir cell clusters (the midline raphe, and the dorsolateral and ventrolateral serotonergic cell groups) and the circumventricular cerebrospinal fluid-contacting neurons in the taenia choroidea (in the caudal brainstem), recessus infundibuli and paraventricular organ (in the hypothalamus) were shown to co-express 5-HT immunoreactivity. However, heavily labeled TPH-ir cell clusters were observed in the nucleus premamillaris (PMM), in the stratum cellulare internum (SCI), in the nucleus paraventricularis magnocellularis (PVN) and in the medial border of the nucleus dorsomedialis anterior thalami (DMA). Double-labeling experiments indicated that none of these medial hypothalamic TPH-ir cells were immunoreactive to 5-HT. These cells correspond to dopamine- and melatonin-containing neurons previously found in the avian hypothalamus, and appear to be comparable to the mammalian TPH-ir hypothalamic A11-A13 catecholaminergic somata, suggesting that they may be a conserved attribute in the amniote medial hypothalamus.

Behavioural and electroencephalographic effects of systemic injections of 8-OH-DPAT in the pigeon (Columba livia).

The effects of systemic injections of the 5HT(1A) receptor agonist 8-OH-DPAT on the spontaneous ingestive, maintenance, locomotor and sleep-like behaviours, and the sleep/waking-related hippocampal electrographic activity were investigated in pigeons. 8-OH-DPAT (0.06, 0.2, 0.6 or 2.0mg/kg) was found to dose-dependently reduce food and water intake, acutely (in the first 3h) and 24h after treatment, during both low-activity morning hours (starting at 10:00 h) and high-activity evening hours (starting at 14:00 h). Automated 24h records of food and water intake indicated that hypophagic effects can last up to 18 h after injection. Duration and incidence of sleep-like postures increased at all doses, in both morning and afternoon. These effects were associated with decreases in exploratory and preening activities. The 8-OH-DPAT-induced hypnogenic, hypophagic and hypodipsic effects tended to be more intense in the morning than in the afternoon-trials. Pretreatment with WAY 100635 (a 5-HT(1A) antagonist; 0.6 mg/kg) eliminated all of these 8-OH-DPAT-induced effects. WAY 100635 failed to affect feeding when injected alone, but decreased frequency of sleep-like responses and increased the latency to the first sleep-like episode. Hippocampal EEG tracings after 8-OH-DPAT injections (0.6 or 2.0mg/kg) indicated that the hypnogenic effects are associated with a specific increase in the frequency and duration of slow wave sleep. Power density analysis of the hippocampal EEG failed to show differences between 8-OH-DPAT-induced sleep and the sleep occurring after vehicle injections, indicating that it may be electrographically similar to diurnal sleep episodes in the pigeon. These data suggest that while 5-HT(1a) receptor-mediated mechanisms play crucial roles in ingestive and sleep/waking behaviours in mammals and birds, their action upon these states shows substantial inter-taxon variance.