Proliferative Defects and Formation of a Double Cortex in Mice Lacking Mltt4 and Cdh2 in the Dorsal Telencephalon

Radial glial cells (RGCs) in the ventricular neuroepithelium of the dorsal telencephalon are the progenitor cells for neocortical projection neurons and astrocytes. Here we showthatthe adherens junction proteins afadin and CDH2 are criticalforthe control of cell proliferation in the dorsal telencephalon and for the formation of its normal laminar structure. Inactivation of afadin or CDH2 in the dorsal telenceph-alon leads to a phenotype resembling subcortical band heterotopia, also known as “double cortex,” a brain malformation in which heterotopic gray matter is interposed between zones of white matter. Adherens junctions between RGCs are disrupted in the mutants, progenitor cells are widely dispersed throughout the developing neocortex, and their proliferation is dramatically increased. Major subtypes of neocortical projection neurons are generated, but their integration into cell layers is disrupted. Our findings suggest that defects in adherens junctions components in mice massively affects progenitor cell proliferation and leads to a double cortex-like phenotype.

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.

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.

Proliferative Defects and Formation of a Double Cortex in Mice Lacking Mltt4 and Cdh2 in the Dorsal Telencephalon

Radial glial cells (RGCs) in the ventricular neuroepithelium of the dorsal telencephalon are the progenitor cells for neocortical projection neurons and astrocytes. Here we showthatthe adherens junction proteins afadin and CDH2 are criticalforthe control of cell proliferation in the dorsal telencephalon and for the formation of its normal laminar structure. Inactivation of afadin or CDH2 in the dorsal telenceph-alon leads to a phenotype resembling subcortical band heterotopia, also known as “double cortex,” a brain malformation in which heterotopic gray matter is interposed between zones of white matter. Adherens junctions between RGCs are disrupted in the mutants, progenitor cells are widely dispersed throughout the developing neocortex, and their proliferation is dramatically increased. Major subtypes of neocortical projection neurons are generated, but their integration into cell layers is disrupted. Our findings suggest that defects in adherens junctions components in mice massively affects progenitor cell proliferation and leads to a double cortex-like phenotype.

Effects of the basic fibroblast growth factor and its anti-factor in the healing and collagen maturation of infected skin wound

PURPOSE: The infection is one of the main factors that affect the physiological evolution of the surgical wounds. The aim of this work is to evaluate the effects of fibroblast growth factor (FGFâ) and anti-FGFâ in the healing, synthesis and maturation of collagen when topically used on infected skin wounds of rats. METHODS: An experimental study was perfomed in 60 male Wistar rats. All animals were divided in two groups (A and B). Each group was divided in three subgroups A1, B1; A2, B2 and A3, B3. After anesthesia with pentobarbital, two open squared wounds (1cm2), 4cm distant to each other, were done in the dorsal skin of all the rats. In group A (n=30) the wounds were contaminated with multibacterial standard solution, and in group B(n=30) the wounds were maintained sterile. These wounds were named F1 (for inflammation analysis) and F2 (for collagen study). The open wounds of A1 and B1 rats were topically treated with saline solution, A2 and B2 were treated with FGFâ and subgroups A3 and B3 were treated with FGFâ and anti-FGFâ. The rats were observed until complete epitelization of F2 wounds for determination of healing time and the expression of types I and III collagen, using Picro Sirius Red staining. Inflammatory reaction in F1 wounds was studied using hematoxilineosin staining. The three variable was measured by the Image Pro-Plus Média Cybernetics software. The statistical analysis was performed by ANOVA and Tukey test, considering p<0.05 as significant. RESULTS: It was observed that infection retarded significantly (p<0.05) the time of wound scarring and the topical application of FCFb reverted the inhibition of healing caused by bacteria. The inflammatory reaction was greater in the subgroup B2 than in B1 and A3, and the difference was significant (p<0.05). It was observed greater expression of type I collagen in all the subgroups treated with FCFb, when compared with the untreated subgroups. Type III collagen was significantly decreased in wounds of B3 rats, comparing to the other subgroups. CONCLUSIONS: The FCFb accelerated the healing of open infected wounds and contributed with maturation of collagen, enhancing the type I collagen density. The anti-FCFb antibody was able to attenuate the production of both type I and III collagen

A nicotina atua como fator deletério na reparação da parede abdominal

Objetivo: Avaliar o efeito da nicotina na cicatrização da camada musculoaponeurótica da parede abdominal. Métodos: Estudo experimental em que foram usados 16 ratos da raça Wistar pesando em média 210±8g, separados aleatoriamente em 2 grupos de 8. Nos animais do grupo A foi implantado disco de nicotina (Nicotinel Ò) na dose de 5mg/Kg de peso/dia no subcutâneo da região dorsal, trocado a cada dois dias, a partir do 5º dia antes da operação em que foi feita laparotomia mediana de 5 cm, até o 10o dia de observação. No grupo B (controle) foram usados discos de celulose com o mesmo diâmetro. Tubo de silicone multiperfurado foi implantado no subcutâneo a 1cm da lesão da parede abdominal. A camada musculoaponeurótica e a pele foram suturadas com fio de nylon 5-0. No 10º dia pós-operatório foi colhido 1ml de líquido seroso do tubo de silicone por punção percutânea para dosagem de pO2 e os animais receberam dose letal de anestésico. Foi ressecado um segmento da camada musculoaponeurótica com 2cm de largura para tensiometria, em seguida processado e corado em HE e tricrômico de Masson para análise quantitativa dos dados histopatológicos em sistema digitalizado. A análise estatística foi feita pelo ANOVA e teste Newman-Keuls, com significância 0,05. Resultados: No grupo A a pO2 do líquido tecidual atingiu o valor 17,75±3,4 mmHg e no grupo B (controle) a pO2 = 40,75±6,4 mmHg (p<0,01). A resistência à tensão apresentou o valor de 728,5±161,75gf no grupo A e 1241,6±232gf no grupo de controle (p<0,01), coincidindo com os achados da pO2. A densidade média dos elementos histopatológicos estudados foi de 105±17,1 nos animais do grupo A e 146,2±8,8 no grupo B (p<0,01). Conclusão: após avaliação da pO2 tecidual, tensiometria e histopatologia, concluiu-se que a nicotina por via subcutânea exerce efeito deletério sobre a cicatrização de lesões da parede abdominal de ratos

Sonic hedgehog signaling regulates mode of cell division of early cerebral cortex progenitors and increases

The morphogen Sonic Hedgehog (SHH) plays a critical role in the development of different tissues. In the central nervous system, SHH is well known to contribute to the patterning of the spinal cord and separation of the brain hemispheres. In addition, it has recently been shown that SHH signaling also contributes to the patterning of the telencephalon and establishment of adult neurogenic niches. In this work, we investigated whether SHH signaling influences the behavior of neural progenitors isolated from the dorsal telencephalon, which generate excitatory neurons and macroglial cells in vitro. We observed that SHH increases proliferation of cortical progenitors and generation of astrocytes, whereas blocking SHH signaling with cyclopamine has opposite effects. In both cases, generation of neurons did not seem to be affected. However, cell survival was broadly affected by blockade of SHH signaling. SHH effects were related to three different cell phenomena: mode of cell division, cell cycle length and cell growth. Together, our data in vitro demonstrate that SHH signaling controls cell behaviors that are important for proliferation of cerebral cortex progenitors, as well as differentiation and survival of neurons and astroglial cells.

Avaliação da toxicidade aguda do inseticida metomil e o seu efeito sobre a atividade da acetilcolinesterase do peixe Danio rerio

Methomyl (Lannate®) is an insecticide from the carbamate group, frequently used in pest control in various types of crops. This compound works inhibiting the activity of the enzyme acetylcholinesterase. The use of physicochemical and ecotoxicological analysis is the most efficient strategy for the correct characterization and control of residues of metomil. The main objectives of this study were to evaluate the acute toxicity of methomyl in 96 hours of exposure and, through a sublethal assay of 5 hours, to assess its effect on the activity of acetylcholinesterase present in brain and squeletic dorsal muscle of the Danio rerio fish. The results showed that the LC50-96 found to D. rerio was 3.4 mg/L and it was found through the average of four definitive tests. In vitro assays were used to test the inhibitory action of methomyl directly over soluble AChE, extracted from the squeletic dorsal muscle, with maximum inhibition of 68.57% to the insecticide concentrations of 0.2 mg/L. In sublethal tests with D. rerio, inhibitory effect of methomyl was found over the soluble form of AChE in the squeletic dorsal muscle, both in one and five hours of fish exposure to the insecticide. In both period, the average values of inhibition were around 61%. In the same condition, no significant inhibitory effect of methomyl soluble and membrane AChE of the D. rerio was observed in the 0.42, 0.85, 1.70 and 2.50 mg/L concentrations and in both times of fish exposure

Biologia alimentar e morfohistologia do tubo digestório do mussum, synbranchus marmoratus, bloch (osteichthyes: synbranchidae) no açude Marechal Dutra Gargalheiras , localizado no semi-árido brasileiro

Food habits and morpho-histology of the digestive tract of marbled swamp eel, Synbranchus marmoratus (Block, 1917) were investigated. The fish samples were captured during August, 2007 to July, 2008 in the Marechal Dutra reservoir, Acari, Rio Grande do Norte. The rain fall data was obtained from EMPARN. The fish captured, were measured, weighed, dissected, eviscerated and individual stomach weights were registered. The stomach contents analyses were carried out based on volumetric method, points, frequency of occurrence and applying the Index of Relative Importance. The degrees of repletion of the stomachs were determined besides the Index of Repletion relating to feeding activity variations and frequency of ingestion during the rainy and dry seasons. The rainfall varied from 0 mm a 335 mm with a mean value of 71.62 mm. Highest rainfall of 335.5 mm was registered in March, 2008 and August to December was the dry period. During the dry period the study species presented high degrees of repletion of the stomachs, with a peak value in the month of September (mean = 4.54; ± SD = 0.56). The minimum mean value of = 3.99 ± SD = 0.25 was registered in the month of May during the rainy period. The stomach contents of S. marmoratus registered show that this fish prefers animals, 78.22% of crustaceans 2.85% of mollusks, 3.25% of fish, 1.4% of insects and 13.5% of semi-digested organic matter, thus characterizing the study species as a carnivore with a preference for crustaceans. The morpho-histological aspects of the digestive tract of S. marmoratus indicate that the mouth is terminal adapted to open widely, thin lips with taste buds, small villiform teeth forming a single series on maxillas, four pairs of branchial arches with short and widely spaced branchial rays. The oesophagus is short and cylindrical with a small diameter. The oesophagus wall is thick with mucas surface and internal parallel folds. The stomach is retilinical in form, presenting cardiac, caecal and pyloric portions. The caecal portion is long and is intermediary in position between the cardiac and pyloric portions. The cardiac portion of the stomach is short and cylindrical formed of simple epithelial cylindrical mucus cells. The caecal portion is long with narrow walls, a big cavity and smaller folds which give rise to gastric glands. The phyloric portion has no glands and primary or secondary mucas folds. The morphohistological aspects of the digestive tract of S. marmoratus indicate its adaptation to a carnivorous feeding habit

Análise funcional do design das órteses para rizartrose

The purpose of this paper is to identify the utilization of two models of orthotics used in the conservative treatment of rizartrose in patients/consumers that were referred by the medical clinic to the LAI (Laboratory of Integrated Accessibility). It describes this group s perception related to the functional aspects of the two models, evaluates the value of the product attributed by the patient/consumer and describes the importance attributed to each model. The research s objective consists in identifying which model this is preferred from the point of view by the patients that use either the dorsal or ventral models used in the conservative treatment of rizartrose . This work is then based on the exploratory research through the investigation of the bibliography available and in the research field. The research field used the case study methodology to investigate the phenomenon in the real context. The data were collected using a patient questionnaire during two times after the use of the dorsal orthotic and after the use of the ventral orthotic. The questionnaire was used to identify the relationship between the patients/consumers and the two different types of orthotics. A general descriptive analysis was used in order to verify the patient s abilities executing certain activities daily living using the dorsal and ventral orthotics, the product s functional aspects and its value from the patient s/consumer s point of view. The results show that the preference for the dorsal or ventral models is determined by the individual needs of each person. The results also show that other variables, such as the use of the orthotic on the dominant or non-dominant hand, play a role in the preferred model by the patient/consumer and need to be further investigated

Plasticidade induzida por treinamento locomotor na medula espinal intacta em ratos: correlatos morfológicos

The locomotion is one of the most important capabilities developed by the animals, whose improvement is dependent on several neural centers, including the spinal cord. This activity promotes a lot of spinal modifications that enable it to adapt and improve their connections. This study aimed to observe the morphological changes occurring in the spinal cord after locomotor training in intact rats. For that we used male Wistar rats, which were submitted to locomotor training in wheel activity in protocols 1, 3 and 7 days (30min/day), and the results were compared to a control group not subjected to exercise. Coronal sections of 40 μm of the lumbosacral spinal cord were subjected to immunohistochemical techniques anti-Egr1, anti-NMDA and anti-SP, to characterize the spinal plasticity related to these substances. Egr1-immunoreactive cells were increased in all laminas, essentially in those more intensely activated by locomotion, laminas IV-X levels L4-S3. All observed sections expressed NMDA-immunoreactivity. Analysis of SP in the spinal dorsal horn resulted no significant variations of this neuropeptide related to locomotion. The results suggest that locomotor training provides synaptic plasticity similar to LTP in all laminas of the lumbosacral spinal cord, in different intensities. However, the SP appears do not participate of this process in the spinal dorsal horn. This work will contribute for consolidating and characterization of synaptic plasticity in the spinal cord

Caracterização dos acoplamentos fase-amplitude na região CA1 do hopocampo

Brain oscillation are not completely independent, but able to interact with each other through cross-frequency coupling (CFC) in at least four different ways: power-to-power, phase-to-phase, phase-to-frequency and phase-to-power. Recent evidence suggests that not only the rhythms per se, but also their interactions are involved in the execution of cognitive tasks, mainly those requiring selective attention, information flow and memory consolidation. It was recently proposed that fast gamma oscillations (60 150 Hz) convey spatial information from the medial entorhinal cortex to the CA1 region of the hippocampus by means of theta (4-12 Hz) phase coupling. Despite these findings, however, little is known about general characteristics of CFCs in several brain regions. In this work we recorded local field potentials using multielectrode arrays aimed at the CA1 region of the dorsal hippocampus for chronic recording. Cross-frequency coupling was evaluated by using comodulogram analysis, a CFC tool recently developted (Tort et al. 2008, Tort et al. 2010). All data analyses were performed using MATLAB (MathWorks Inc). Here we describe two functionally distinct oscillations within the fast gamma frequency range, both coupled to the theta rhythm during active exploration and REM sleep: an oscillation with peak activity at ~80 Hz, and a faster oscillation centered at ~140 Hz. The two oscillations are differentially modulated by the phase of theta depending on the CA1 layer; theta-80 Hz coupling is strongest at stratum lacunosum-moleculare, while theta-140 Hz coupling is strongest at stratum oriens-alveus. This laminar profile suggests that the ~80 Hz oscillation originates from entorhinal cortex inputs to deeper CA1 layers, while the ~140 Hz oscillation reflects CA1 activity in superficial layers. We further show that the ~140 Hz oscillation differs from sharp-wave associated ripple oscillations in several key characteristics. Our results demonstrate the existence of novel theta-associated high-frequency oscillations, and suggest a redefinition of fast gamma oscillations

Interferindo com oscilações de alta frequência no hipocampo epiléptico: consequências para as crises espontâneas

Conselho Nacional de Desenvolvimento Científico e Tecnológico

Análise quantitativa das células CD4 e sua importância no diagnóstico de lesões orais em pacientes HIV positivos

The oral manifestations due to HIV infection are, a lot of times, the first clinical signs of the disease. These injuries may also function as beepers and sentries of the curse and progression of the HIV infection and AIDS. The objective of this work was to evaluate the prevalence of the oral injuries in HIV positive patients, relating them with the CD4+ cells counting and the viral load in patients from the Hospital of Infected contagious Gizelda Trigueiro in Natal-RN. One hundred and one patients were evaluated, where after the clinical exam of the oral cavity, these ones were conducted to the peripheral blood collection for the counting of CD4+ lymphocytes. We observed a prevalence of 25,6%, that is, 31 cases. The Oral Candidiasis was the most commum injure, followed by Oral Hairy Leukoplakia, linear gingival erytema, lips herpes, gingivitis and periodontitis - HIV. The average counting of cells CD4+ of the injury carrying patients was of 250 cells/mm3. We did not observe relation between the presence of injuries and the viral load of the individuals