Behavioral and metabolic effects of central injections of orexins/hypocretins in pigeons (Columba livia).

In the present study, the acute behavioral and ingestive effects of ICV injections of mammalian orexin-A (ORXA; vehicle, 0.2, 0.6 or 2 nmol) and of orexin-B (ORXB; vehicle, 0.2, 0.6 or 2 nmol), as well as possible long-term effects (through 24 h of continuous intake monitoring after 0.6 nmol of ORXA or ORXB) of these treatments in food/water intake and in blood levels of metabolic fuels (free fatty acids and glucose, after 0.2 or 0.6 nmol of ORXA) were examined in adult male pigeons. Both ORXA and ORXB treatments failed to produce acute (1-3 h) or long-term effects on feeding and drinking behaviors, and did not change blood free fatty acids and glucose 15 and 30 min after treatments, as compared to vehicle-treated animals. However, ORXA (but not ORXB) treatments evoked a dose-related, intense increase in exploratory behaviors, associated to reduced time spent in alert immobility and sleep-typical postures. These data substantiate the lack of orexigenic effects of ORXs in avian species, and suggest that an important role in vigilance control may represent a conserved functional attribute of orexinergic circuits in vertebrates.

Influence of enrichment on behavioral and neurogenic effects of antidepressants in Wistar rats submitted to repeated forced swim test.

Repeated forced swimming test (rFST) may detect gradual effects of antidepressants in adult rats. Antidepressants, as enrichment, affected behavior and neurogenesis in rats. However, the influence of enrichment on behavioral and neurogenic effects of antidepressants is unknown. Here, effects of antidepressants on rFST and hippocampal neurogenesis were investigated in rats under enriched conditions. Behaviors of male Wistar rats, housed from weaning in standard (SE) or enriched environment (EE), were registered during rFST. The rFST consisted of 15min of swimming (pretest) followed by 5min of swimming in the first (test), seventh (retest 1) and fourteenth (retest 2) days after pretest. One hour before the test, rats received an intraperitoneal injection of saline (1ml/kg), fluoxetine (2.5mg/kg) or imipramine (2.5 or 5mg/kg). These treatments were performed daily until the day of the retest 2. After retest 2, rats were euthanized for the identification of markers for neurogenesis in the hippocampus. Fluoxetine or imipramine decreased immobility in retests 1 and 2, as compared to saline. EE abolished these differences. In EE, fluoxetine or imipramine (5mg/kg) reduced immobility time in retest 2, as compared to the test. Independent of the housing conditions, fluoxetine and imipramine (5mg/kg) increased the ratio of immature neurons per progenitor cell in the hippocampus. In summary, antidepressants or enrichment counteracted the high immobility in rFST. Enrichment changed the effects of antidepressants in rFST depending on the type, and the dose of a substance but failed to change neurogenesis in control or antidepressant treated-rats. Effects of antidepressants and enrichment on rFST seemed neurogenesis-independent.

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.

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.

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.

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.

Repercussões morfológicas e funcionais do exercício sobre a regeneração nervosa periférica / Morphological and functional repercussion on peripheral nerve regeneration after exercises

INTRODUÇÃO: A indicação de exercícios físicos como abordagem terapêutica após lesões nervosas periféricas, apesar de resultados clínicos favoráveis, ainda não é consenso na literatura pertinente. Objetivos: Este estudo buscou avaliar a interferência do exercício em roda motorizada no processo de regeneração nervosa periférica. MATERIAIS E MÉTODOS: Foram utilizados 48 ratos Wistar divididos proporcionalmente em quatro grupos de 12 ratos cada (seis controles e seis experimentais). Após treinamento, os ratos foram submetidos à axonotmese do nervo ciático direito. Todos os grupos iniciaram o regime de exercícios diários 24 horas após a lesão, porém com durações diferenciadas (G1 por 28 dias; G2 por 21 dias, G3 por 14 dias e G4 por sete dias). O exercício teve tempo e velocidade ajustados de acordo com o tempo de tratamento para cada grupo. RESULTADOS: Por meio do cálculo do Índice Funcional do Ciático, os animais submetidos ao exercício apresentaram sinais de atraso na recuperação funcional da marcha, em comparação a animais controles. Na análise histológica dos nervos lesionados, observou-se que animais exercitados apresentaram sinais de preservação dos axônios no coto proximal e de degeneração no coto distal, e os animais exercitados por somente sete dias não apresentaram a mesma característica degenerativa no coto distal. A aplicação de exercícios ativos e sem carga em animais que sofreram axonotmese interfere negativamente no processo de recuperação funcional de alguns parâmetros da marcha. CONCLUSÃO: Apesar de o exercício ativo não ter interferido na manutenção da viabilidade neuronal nos sítios proximais à lesão, sua continuidade prejudicou a viabilidade dos cotos neurais distais, provável causa do atraso na recuperação funcional da marcha.

INTRODUCTION: This study aimed to investigate if exercise in motorized wheel can affect the process of peripheral nerve regeneration in a rat model of axonotmesis. MATERIAL AND METHODS: Forty-eight Wistar rats were divided equally into four groups containing six experimental and six control subjects. After proper training, the rats underwent right sciatic nerve axonotmesis. All groups started the exercises, in a daily basis, 24 hours after nerve injury, but at different durations (28 days for G1, 21 days for G2, 14 days for G3 and seven days for G4). Both duration and speed of the exercises were gradually adjusted during the experiments. RESULTS: The Sciatic Function Index revealed that exercised animals showed clear signs of delayed functional gait recovery. The histological analysis of injured nerves revealed that exercised animals showed signs of proximal stump preservation and degenerative signs in the distal stump. In other hand, exercised animals during seven days only showed axonal preservation in the distal stump. CONCLUSION: Based on these results, we believe that active exercises should be avoided, at least during the peripheral nerve regeneration process, i.e., during first 21 days after axonotmesis.".