Alterações da default mode network provocadas pela ingestão de Ayahuasca investigadas por Ressonância Magnética Funcional

Ayahuasca is psychotropic beverage that has been used for ages by indigenous populations in South America, notably in the Amazon region, for religious and medicinal purposes. The tea is obtained by the decoction of leaves from the Psychotria viridis with the bark and stalk of a shrub, the Banisteriopsis caapi. The first is rich in N-N-dimethyltryptamine (DMT), which has an important and well-known hallucinogenic effect due to its agonistic action in serotonin receptors, specifically 5-HT2A. On the other hand, β-carbolines present in B. caapi, particularly harmine and harmaline, are potent monoamine oxidase inhibitors (MAOi). In addition, the tetrahydroharmine (THH), also present in B. caapi, acts as mild selective serotonin reuptake inhibitor and a weak MAOi. This unique composition induces a number of affective, sensitive, perceptual and cognitive changes in individuals under the effect of Ayahuasca. On the other hand, there is growing interest in the Default Mode Network (DMN), which has been consistently observed in functional neuroimaging studies. The key components of this network include structures in the brain midline, as the anterior medial frontal cortex, ventral medial frontal cortex, posterior cingulate cortex, precuneus, and some regions within the inferior parietal lobe and middle temporal gyrus. It has been argued that DMN participate in tasks involving self-judgments, autobiographical memory retrieval, mental simulations, thinking in perspective, meditative states, and others. In general, these tasks require an internal focus of attention, hence the conclusion that the DMN is associated with introspective mental activity. Therefore, this study aimed to evaluate by functional magnetic resonance imaging (fMRI) changes in DMN caused via the ingestion of Ayahuasca by 10 healthy subjects while submitted to two fMRI protocols: a verbal fluency task and a resting state acquisition. In general, it was observed that Ayahuasca causes a reduction in the fMRI signal in central nodes of DMN, such as the anterior cingulate cortex, the medial prefrontal cortex, the posterior cingulate cortex, precuneus and inferior parietal lobe. Furthermore, changes in connectivity patterns of the DMN were observed, especially a decrease in the functional connectivity of the precuneus. Together, these findings indicate an association between the altered state of consciousness experienced by individuals under the effect of Ayahuasca, and changes in the stream of spontaneous thoughts leading to an increased introspective mental activity

Efeitos da senescência no núcleo supraquiasmático do sagui (Callithrix Jacchus): plasticidade morfológica e neuroquímica

The circadian timing system (CTS) is responsible for the generation and synchronization and the suprachiasmatic nucleus (SCN) of the hypothalamus has been described as the major circadian pacemaker in many mammalian species. The internal temporal organization managed by SCN is disturbed with aging bringing many pathological disorders that range from loss of complex cognitive performance to simple physiological functions. Therefore, our aim was perform a comparative study of the morphological aspects and neurochemical composition in the SCN of marmosets (Callithrix jacchus) adults and older using immunohistochemical techniques. We found morphometric and neurochemical changes in th SCN o folder animals in comparison to adults, among these a possible decreased in retinal projection to the SCN of older animals, found through a decline in CTB immunostaining, which can occur due atrophy and/or decreasing of fibers from the retinohypothalamic tract (RHT). The Klüver-Barrera histological technique strongly suggests a decrease in those fibers from RHT. Also, by means of a morphometric study, it is found a atrophy and numerical decline of neurons in SCN of aged animals, investigated by Nissl technique, and immunostaining with NeuN and calbindin. Relative optical density (ROD) analysis were used to evaluate the expression of some neurochemical components in SCN, such as GFAP expression, which was increased in older, result that indirectly reinforces that morphological changes occurs due the aging; the vasoactive intestinal polipeptide (VIP) showed no expression alteration in SCN of older animals; the serotonin (5-HT) was descreased in the dorsomedial portion of the SCN, and neurpeptide Y (NPY) apparently also decrease due to the increase of age. Many of these modifications were seen in other animals, such as rodents, human primates and non-human primates. These data about marmoset comes to add new information of the effect of aging on structures responsibles for the circadian rhytmicity, and that some behavioral changes controlled by th SCN, and founded in aged animals, may be caused by these morphological and neurochemical changes. Although some results have been quantitatively negative, qualitatively all analysis show significant change comparing adult and older animals, perhaps due to a low sampling number. In conclusion, the marmoset presents several morphological and neurochemical changes in the SCN of aged animals compared to adults, which may result in behavioral changes that favor pathology aging related

Centros rombencefálicos de processamento auditivo do sagui (Callithrix jacchus): uma análise citoarquitetônica e neuroquímica

The auditory system is composed by a set of relays from the outer ear to the cerebral cortex. In mammals, the central auditory system is composed by cochlear nuclei, superior olivary complex, inferior colliculus and medial geniculate body. In this study, the auditory rombencephalic centers, the cochlear nuclear complex and the superior olivary complex were evaluated from the cytoarchitecture and neurochemical aspects, thorough Nissl staining and immunohistochemical techniques to reveal specific neuron nuclear protein (NeuN), glutamate (Glu), glutamic acid decarboxilase (GAD), enkephalin (ENK), serotonin (5-HT), choline acetyltransferase (ChAT) and calcium-binding proteins calbindin (CB), calretinin (CR), and parvalbumin (PV). The common marmoset (Callithrix jacchus), a little native primate of the Brazilian atlantic forest was used as an experimental animal. As results, it was noted that the cochlear nuclear complex is composed by anteroventral, posteroventral and dorsal nuclei, and the superior olivary complex is constituted by the lateral and medial superior olivary nuclei and the trapezoid body nucleus. Glu, GAD, ENK, ChAT, CB, CR, PV-immunoreactive cells, fibers and terminals besides besides only 5-HT terminals were found unhomogeneously in all nuclei, of both complex. The emerging data are discussed in a comparative and functional context, and represent an important contribution to knowledge of the central auditory pathways in the common marmoset, and then in primates

A Zona Incerta no sagüi (Callithrix jacchus): Análise Citoarquitetônica, Neuroquímica e Projeção Retiniana

The retinal projections in mammals usually reach, classically, three major functional systems: the primary visual system, the accessory optic system, and the circadian timing system. But the retinal projections also reach areas classically considered non-visual, one of which groups the neurons of the zona incerta (ZI), target this study. The primary visual system includes thalamic lateral geniculate complex is formed by the dorsal lateral geniculate nucleus, intergeniculate leaflet and the ventral lateral geniculate nucleus and other Components. The accessory optic system is composed of the small nuclei: nuclei terminal dorsal, lateral, medial and the interstitial nucleus of the superior posterior fasciculus. These nuclei are involved in visuo-motor activities. The circadian timing system is comprised of the suprachiasmatic nucleus of the hypothalamus, that act as master circadian pacemaker, entraining pathways and efferents pathways to the efectors, and the intergeniculate leaflet, that seems to act as a modulator of the pacemaker. The retinal projections too reach classically considered non-visual areas, including the zona incerta. This region is localized in the ventral thalamus and has been implicated in various functional properties including nociceptive and somatosensory processing, motor response, sociosexual behaviour, feeding and drinking, in symptoms of neurodegenerative diseases, arousal and attention. It also displays connection with several areas of central nervous system. The aim of this study was characterize the retinal projection in the zona incerta of Callithrix jacchus (sagüi), a primate of the New World through the anterograde axonal transport of the cholera toxin subunit b and analyze the citoarchicteture using Nissl and NeuN, and neurochemical substances such as serotonin, GABA, VIP, VP, GFAP and binding-calcium proteins. The zona incerta showed a different division of the literature in citoarquitetura, both by means of Nissl as neurochemical by NeuN, with a subdivision ventrolateral and dorsomedial. The neurochemical to the other substances corroborate with this subdivision. The GFAP was almost completely negative for the zona incerta, result non evidenced in previous studies yet. The 16 retinal projection in sagüi, unlike other primates and rodents, reached the caudal portion only. This work helps to make further studies are conducted based on this subdivision and the localization of the neurochemical substances associated with possible behaviors that the zona incerta is involved

O Núcleo genuíno lateral dorsal do tálamo do sagüi (callithrix jacchus): Pprojeção retiniana, caracterização citoarquitetônica e neuroquimica da principal estação visual primária.

The thalamus plays an important role in the sensorial processing information, in this particular case, the visual information. Several neuronal groups have been characterized as conductors and processors of important sensorial information to the cerebral cortex. The lateral geniculate complex is one to them, and appears as a group very studied once it is responsible, in almost all totality, for the processing of visual information. Among the nuclei that constitute the lateral geniculate complex we highlight the dorsal lateral geniculate nucleus of the thalamus (DLG), the main thalamic relay for the visual information. This nucleus is located rostral and lateral to medial geniculate nucleus and ventral to thalamic pulvinar nucleus in most of the mammals. In the primates humans and non-humans, it presents as a laminate structure, arranged in layers, when observed in coronal sections. The objective of this work was to do a mapping of the retinal projections and a citoarchictetonic and neurochemical characterization of DLG in the marmoset (Callithrix jacchus), a New World primate. The retinal projections were traced by anterograde transport of subunit b of cholera toxin (CTb), the citoarchicteture was described by Nissl method, and to neurochemical characterization immunohistochemicals technical were used to examine the main neurotransmitters and neuroatives substances present in this neural center. In DGL of marmoset thalamus, in coronal sections labeled by Nissl method, was possible to visualize the division of this nucleus in four layers divided in two portions: magnocellular and parvocellular. The retinal projections were present being visualized fibers and terminals immunorreactives to CTb (IR-CTb) in the DLG ipsilateral and contralateral. And through the immunohistochemicals techniques was observed that DLG contain cells, fibers and/or terminals immunoreactives against neuronal nuclear protein, subunits of AMPA 15 glutamate receptors (GluR1, GluR2/3, GluR4), choline acetyltransferase, serotonin, glutamic acid decarboxylase, binding calcium proteins (calbindin, parvalbumin and calretinin), vasopressin, vasoactive intestinal polypeptide, and an astrocyte protein, glial fibrillary acidic protein.

Caracterização citoarquitetônica, neuroquímica e de aferência óptica do complexo parabraquial do sagui (Callithrix jacchus)

The parabrachial complex (PB) is an area of the brainstem responsible for the processing and transmission of essential physiologic information for the survival of the organisms. This region is subdivided in approximately nine subregions, considering morphology, cytoarchitectural and functional characteristic. Its neurons have an extensive network of connections with other regions of the nervous system. The objective in this work was to map the retinal projection to the PB and make a citoarchitectonic and neurochemical characterization of this region in the common marmoset (Callithrix jacchus), a primate of the New World. The retinal projections were mapped by anterograde transport of the choleric toxin subunit b (CTb). The citoarchitecture was described through the Nissl method, and the neurochemical characterization was made through immunohistochemical technique to the some neurotransmitters and neuroactives substances present in this neural center. In marmoset PB, in the coronal sections labeled by Nissl method, we found a similar pattern to that evidenced in other animal species. The immunoreactivity against CTb was verified in the PBMv in fibers/terminal, characterizing such as retinal innervations in this area. The immunohistochemical technique reveled that the PB contain cells, fibers and/or terminals immunoreactives to the neuronal nuclear protein, Choline acetyl transferase, nitric oxide synthase, serotonin, enkephalin, substance P, Calcium-binding proteins (calbindin, calretinin e parvalbumin), and glial fibrillary acidic protein. The histochemical technique reveled cells and fibers NADPH-diaphorase reactive. Each one of those substances presented a characteristic pattern of demarcation in PB, and some serve as specific markers of subregions

Delimitação dos grupamentos serotoninérgicos/núcleos da rafe do mocó (kerodon rupestris): citoarquitetura e imunoistoquímica para serotonina

Serotonin or 5-hydroxytryptamine (5-HT) is a substance found in many tissues of the body, including as a neurotransmitter in the nervous system, in which may exert varied post-synaptic actions. Inside the neuro-axis, the location of 5-HT neurons is almost restricted to the raphe nuclei of the brainstem, such that 5-HT-immunoreactivity can be considered a marker of the raphe nuclei. The raphe nuclei are located in the brainstem, at or near the midline. The serotonergic groups were originally alphanumerically classified as B1 to B9 towards caudorrostral in rats and can be divided into upper and lower groups. In this study the distribution of serotonergic neurons was studied using immunohistochemistry in the brain of the rock cavy (Kerodon rupestris), a species of rodent endemic to Northeastern Brazil. The cytoarchitectonic location of serotonergic neurons was established in series of adjacent coronal and sagittal sections stained by the Nissl method and immunohistochemistry for 5-HT. Thus, we defined the raphe rostral linear, caudal linear, dorsal, median, and paramedian pontine raphe nuclei, and B9 cluster, constituting the rostral group, and the interpositus, magnus, obscure and palidus, constituting the caudal part of the group, comparable to which has been described for other mammalian species

Análise imunoistoquímica da distribuição de Serotonina, transportador de serotonina e receptores de serotonina no hipotálamo do sagui (callithrix jacchus)

The hypothalamus is a diencephalic portion located around the third ventricle below the hypothalamic sulcus, limited by the optic chiasm, and by the mammillary bodies, acting as a center that integrates behavioral and homeostatic functions. Serotonin is a neurotransmitter produced in limited sites in the midbrain and brain stem, but is distributed throughout the central nervous system and has many functions, acting through specific receptors that are also distributed throughout the nervous system. Using immunohistochemical techniques, the aim of this study was to delineate the hypothalamic nuclei of the marmoset (Callithrix jacchus) and study the distribution of serotonin transporter and serotonin receptors in the hypothalamus of this species. We used the Nissl method to determine the cytoarchitecture of the hypothalamic nuclei, and immunohistochemistry to reveal the presence of NeuN as a method to determine the contours of the hypothalamic nuclei. As a result, we found serotonin containing fibers and terminals throughout the rostrocaudal extent of the hypothalamus, more concentrated in some nuclei, and even absent in some. Like serotonin, serotonin transporter was observed between pre-optic area and tuberal region of the hypothalamus, in densities and distribution similar to serotonin. The 5-HT1A and 5-HT1B receptors were found with minor differences among itselves regarding the disposition and intensity of staining.

Análise citoarquitetônica dos componentes do sistema de temporização circadiana do sagui (Calithrix jacchus), comparada com a inervação das suas principais aferências

The suprachiasmatic nucleus (SCN) of the anterior hypothalamus, together with the intergeniculate leaflet (IGL) of the thalamus are considered the central components of the circadian timing system (CTS) of mammals. This system is responsible for the generation and regulation of circadian rhythms by establishing a temporal organization of physiological processes and behaviors. The neuronal specific nuclear protein (NeuN) has been widely used as a neuronal marker in several studies. Since glial fibrillary acidic protein (GFAP) is a component of intermediate filaments found in the cytoplasm of astrocytes and is commonly used as a specific marker for these cells. This study aims to identify, in the marmoset, the NeuN immunoreactive neurons and glial cells immunoreactive to GFAP, as well as map the major route of photic synchronization of the STC, retinohypothalamic tract (RHT), and identify the indirect pathway to the SCN and pregeniculate nucleus (PGN) - structure homologous to IGL rodents, using immunohistochemical and cytoarchitectonic techniques. Observed in SCN the presence of neurons immunoreactive to NeuN and terminals immunoreactive subunit b of cholera toxin (CTb), neuropeptide Y (NPY) and serotonin (5- HT). In the PGN noted the presence of the NeuN and NPY immunoreactive neurons and the immunoreactive terminals CTb and 5-HT. Astrocytes are present throughout the extent of the SCN and the PGN this New World primate

Padrão de distribuição e caracterização morfológica de fibras serotonérgicas nos núcleos da linha Média/ intralaminares do tálamo do mocó (Kerodon rupestris)

The midline/intralaminar nuclei form a remarkable group of nuclei of the medial and dorsal thalamus. The midline nuclei, in rats, comprises the paratenial nuclei (PT), paraventricular (PV), intermediodorsal (IMD), reuniens (Re) and rhomboid (Rh). The intralaminar nuclei comprises the central medial (CM), paracentral (PC), central lateral (CL) and parafascicular (PF). Such nuclei have dense serotonergic innervation originating from the brainstem, especially from the so-called ascending activation system. These nuclei, in turn, send projections to various cortical and subcortical areas, specifically to limbic areas, which suggests the important role of this neurotransmitter in the limbic circuitry. The aim of this study was to characterize the distribution pattern and morphology of serotonin fibers in the nuclei of the midline and intralaminar thalamic of rocky cavy (Kerodon rupestris), a tipical rodent from brazilizan northeast. To reach this aim we used four rock cavies adults. Following the transcardially perfusion with paraformaldehyde and brain microtomy steps was performed immunohistochemistry for serotonin (5-HT), Nissl technique and subsequent achievement and image analysis to characterize the cytoarchitecture of these nuclei and the serotonergic fibers visualized. An analysis was made of Relative Optical Density (ROD) to semi-quantify the concentration of serotonin fibers in the areas of interest. Thus, we observed a cytoarchitectonic arrangement of these nuclei similar to that found in rats. In case of fibers distribution, those immunoreactive to 5-HT were presented in a higher concentration according as ROD in the midline nuclei relative to intralaminar; Re being the core which has a higher pixel value followed by the PV , Rh, IMD and PT. In intralaminar CL showed higher pixels, followed by nuclei CM, PC and PF. The serotonergic fibers were classified as number of varicosities and axon diameter, therefore find three types of fibers distributed through this nuclear complex: fibers rugous, granular and semi-granular. In PV fibers predominated rugous; in PT fibers predominated granular; IMD, CL and PF fibers were represented by semi-granular and Re, Rh, PC and CM fibers showed granular and semi-granular. Morphological characterization of serotonergic fibers and differences in density between the nuclei may suggest different patterns of synaptic organization of this neurotransmitter beyond confirming his large repertoire functional

O antagonismo do receptor de serotonina do tipo 7 da substância cinzenta periaquedutal dorsal reduz a ansiedade experimental basal, mas não aquela gerada pela retirada do etanol em ratos

Ethanol-dependent individuals who reduce or discontinue its use may present Alcohol Withdrawal Syndrome, which is characterized by unpleasant signs and symptoms, such as anxiety, that may trigger relapses. Ethanol, a psychotropic drug, is able to promote behavioral and neurophysiological changes, acting on different neurotransmitter systems, including the serotonergic, which has also been directly associated with aversive states, including anxiety. This study aimed to investigate the participation of type 7 serotonin receptor (5-HT7) of the dorsal periaqueductal gray (DPAG) on basal experimental anxiety and that caused by ethanol withdrawal. For this, 75-100 days old Wistar rats were subjected to two experiments. On the first one, animals underwent stereotactic surgery for implantation of guide cannulas used for administration of the drug directly into the DPAG. After seven days, the animals received doses of 2.5; 5 and 10 nmols of type 7 receptor antagonist SB269970 (SB) or vehicle intra-DPAG and, ten minutes after, they were exposed to elevated plus maze (EPM). In the following day, the animals were submitted to the same treatment and tested in the open field (OF). In the second experiment, animals received increasing concentrations (2%, 4%, 6%) of ethanol as the only source of liquid diet or water (control group), both with free access to chow. Seventy two hours and ninety six hours after the ethanol withdrawal, animals received SB (2.5 and 5.0 nmols) intraDPAG ten minutes before the test in the LCE and OF, respectively. In experiment 1, the dose of antagonist 10 nmols was able of reversing the anxiety generated by EPM. In the experiment 2, ineffective SB doses on the LCE (2.5 and 5.0 nmol) were not able to reverse the anxiety caused by the ethanol withdrawal in the EPM, although the dose of 2.5 nmols of SB has reversed its hipolocomotor effect in this test. This result suggests that the 5-HT7 receptor is involved in the modulation of the basal experimental anxiety in rats, but not in the anxiety caused by ethanol withdrawal in the DPAG.

Avaliação dos efeitos da retirada do etanol em curto e longo prazo sobre respostas comportamentais relacionadas à ansiedade e sobre células imunorreativas para a serotonina no núcleo dorsal da Rafe em ratas

Ethanol withdrawn individuals present a wealth of signs and symptoms, some of them related with anxiety. To better understand brain areas involved in anxiety caused by ethanol abstinence, preclinical studies have been employing models of ethanol consumption followed by withdrawal in rodents submitted to behavioral tests of anxiety, such as the elevated plus-maze. The aim of this study was to investigate if short- or long-term ethanol withdrawal could alter both anxiety-related behaviors in the elevated plus-maze (EPM) and open field tests and the number of serotonin immunorreactive cels in the dorsal raphe nucleus, a midbrain area associated with anxiety. Female Wistar rats (90 days old) were submitted to increasing concentrations of ethanol (2% for 3 days, 4% for 3 days and 6% for 15 days) as the only source of liquid diet and the control group received water ad libitum. Both groups received food ad libitum. In the behavioral experiments, on 21st day of consumption, ethanol was substituted by water (withdrawal) and 72 h or 21 days after withdrawal animals were submitted to the EPM, where it was evaluated the percentage of time and entries in the open arms and the entries in the enclosed arms during 5 minutes. Twenty and four hours after testing in the EPM, animals were submitted to the open field test for 15 minutes, where the distance traveled by the animals was observed along this period. During the first 5 minutes, the distance traveled, entries and time spent in the center of the test were analyzed. In the immunohistochemistry study, animals were submitted to 21 days of consumption of ethanol followed or not by 72 hours and 21 days of withdrawal previously perfusion, brain tissue preparation and quantification of serotonin dyed cells in the dorsal and caudal portions in the dorsal raphe nucleus. Behavioral data showed that both short- and long-term ethanol withdrawals reduced the open arms exploration in the EPM. In the open field test there were no locomotor activity changes during the total 15 minutes; however, longterm ethanol withdrawal reduced the exploration in the center of the open field during the first 5 minutes. In the immunohistochemistry step, there were no differences, when short- and long-term withdrawn groups were compared with control group; nevertheless, the chronic consumption of ethanol decreased the number of serotonergic immunorreactive cells in the dorsal part of dorsal raphe nucleus. Taken together, results here obtained suggest that both short- and long-term ethanol withdrawals promoted an anxiogenic-like effect that was not related with changes in the serotonin immunorreactivity in the dorsal and caudal parts of the dorsal raphe nucleus.

Organização nuclear do sistema serotonérgico no encéfalo de morcegos Artibeus planirostris

Serotonin or 5-hydroxytryptamine (5-HT) is a substance found in many tissues of the body, including the nervous system acting as a neurotransmitter. Within the neuro-axis, the location of the majority of the 5-HT neurons is superimposed with raphe nuclei of the brain stem, in the median line or its vicinity, so that neuronal 5-HT can be considered a marker of the raphe nuclei. Serotonergic neurons are projected to almost all areas of the brain. Studies show the participation of serotonin in regulating the temperature, feeding behavior, sexual behavior, biological rhythms, sleep, locomotor function, learning, among others. The anatomy of these groups has been revised in many species, including mouse, rabbit, cat and primates, but never before in a bat species from South America. This study aimed to characterize the serotonergic clusters in the brain of the bat Artibeus planirostris through immunohistochemistry for serotonin. Seven adult bat males of Artibeus planirostris species (Microchiroptera, Mammalia) were used in this study. The animals were anesthetized, transcardially perfused and their brains were removed. Coronal sections of the frozen brain of bats were obtained in sliding microtome and subjected to immunohistochemistry for 5-HT. Delimit the caudal linear (CLi), dorsal (DR), median (MnR), paramedian (PMnR), pontine (PNR), magnus (MgR), pallidus (RPA) and obscurus (ROb) raphe nucleus, in addition to the groups B9 and rostral and caudal ventrolateral (RVL/CVL). The serotonergic groups of this kind of cheiroptera present morphology and cytoarchitecture relatively similar to that described in rodents and primates, confirming the phylogenetic stability of these cell clusters.