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Specific neuronal phenotypes within the rostral ventrolateral medulla following cardiovascular deconditioning in ratsZidon, Terese M. January 2008 (has links)
Thesis (M.S.)--University of Missouri-Columbia, 2008. / The entire dissertation/thesis text is included in the research.pdf file; the official abstract appears in the short.pdf file (which also appears in the research.pdf); a non-technical general description, or public abstract, appears in the public.pdf file. Includes bibliographical references.
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Serotonergic axon development in the medulla oblongata in post-natal miceTyagi, Ayushi 08 April 2016 (has links)
Sudden Infant Death Syndrome (SIDS) is the sudden death of an infant younger than one year of age that remains unexplained after a complete investigation. For these infants, many different reasons have been hypothesized as to the cause of these deaths including: inherent vulnerability and improper hypoxic arousal. Studies done in other laboratories have shown that there seems to be a reduction in the levels of the neurotransmitter serotonin (5-HT) in the neurons of the raphe, extra-raphe, and ventral populations along with projection sites of these neurons. The huge implications of 5-HT in the control of respiration, prompted animal model studies to further investigate a potential connection between 5-HT and SIDS. 5-HT deficient mice were engineered by knocking out the Pet-1 transcription factor so that knockout mice only retained 30-40% of their brainstem 5-HT neurons. By comparing these 5-HT deficient Pet-1 knockout mice to wild-type mice, it was demonstrated that 5-HT deficient mice failed to autoresuscitate themselves after repeated bouts of hypoxia. Intriguingly, these mice only experienced an autoresuscitation deficit during a specific time period during development. To further evaluate the pathological development behind this behavior issue, in the current study we utilized mice that have modified Pet-1-Flpe driver, Egr2-Cre driver, along with a knock-in RC::FPSit allele to observe 5-HT development in the brainstem in a mature adult and across the critical period (postnatal days 8 and 13- P8 and P13). The transgenic mouse model Pet1-Krox20 gives us a way of exploring a specific subset of 5-HT neurons that rise from the developmental rhombomeres r3 and r5. The use of the knock-in RC::FPSit allele allows us to view the axonal projections of these specific 5-HT neurons by utilizing the presynaptic marker synaptophysin-GFP. This model (PKSit) will allow us to target 5-HT neurons that are implicated in respiration. We chose to compare two projection targets of the PKSit 5-HT neuron subtype through the vulnerable period of development and mature adult mouse: the Locus Coeruleus (LC) and the Nucleus Tractus Solitarius (NTS). In this study we tested the amount of colabeling between 5-HT and GFP in the LC and NTS at P8, P13, as well as the mature adult. We hypothesize that the LC undergoes significant serotonergic axon development and increases colocalization with GFP labeled axon projections between the ages of P8 and P13. We sliced mouse brains and ran immunofluorescence before taking confocal images. By utilizing ImageJ software to run colocalization analysis on the images obtained, we were able to quantify the amount of 5-HT labeled axon projections that are colocalized with GFP labeled axon projections. The parameters we used to quantify the amount of colocalization include the Pearson's Coefficient (PC), Mander's Coefficient (M1/M2), Cytofluorograms, Costes' Method, and van Steensel's Cross-Correlation Coefficient (CCF). We found that the LC shows significant changes with age in the colocalization of 5-HT with GFP while the NTS does not exhibit significant changes with age. The significant changes found in the LC 5-HT/GFP expression between the ages of P8 and P13 suggest one possible cause of failure of arousal. At P8, this lack of 5-HT colabeling with GFP projections suggests that there is some development occurring, which prevents the proper function of 5-HT. At P13, there is a significant increase in the colabeling of 5-HT with GFP, which indicates that the Pet1-Krox20 lineage is actively using 5-HT. The colocalization studies demonstrate that as the mouse ages, the amount of 5-HT labeling with GFP-synaptophysin in the NTS stays the same. The lack of overlap even in mature adult mice suggests that the expression of 5-HT in GFP labeled projections is not necessary. This colocalization study shows that there is an effect of age on the development of the serotonergic system in the LC, but no effect of age in the NTS. While this demonstrates that there is a critical period of development in relation to the LC, it is only one aspect of why mice pups failed to respond to repeated bouts of hypoxia.
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O bloqueio purinérgico no núcleo retrotrapezóide (RTN) atenua as respostas respiratórias promovidas pela ativação dos quimiorreflexos central e periférico em ratos. / Purinergic receptors blockade in the retrotrapezoid nucleus (RTN) attenuates the central and peripheral chemoreflexes in rats.Barbara Falquetto Barna 19 November 2015 (has links)
O ATP mediando a sinalização purinérgica no bulbo ventrolateral rostral contribui para o controle do quimiorreflexo central e periférico regulando a pressão arterial e a respiração, mediante o envolvimento dos neurônios do núcleo retrotrapezóide (RTN). No entanto, as potenciais contribuições da sinalização purinérgica, no RTN, na função cardiorrespiratória em animais não anestesiados ainda não foram testadas. Mostramos que a injeção de ATP no RTN promoveu aumento cardiorrespiratório por um mecanismo dependente de receptores P2. Mostramos também que o bloqueio de receptor P2 não específico (PPADS), mas não de receptores específicos P2Y (MRS2179), reduziu a resposta ventilatória à hipercapnia (7% CO2) e hipóxia (8 % O2) em ratos não anestesiados. Além disso, a adenosina (ADO) no RTN atenuou o aumento da ventilação induzido por hipercapnia in vivo e o disparo dos neurônios in vitro. Estes resultados demonstram que a sinalização mediada por ATP contribui para o controle respiratório do quimiorreflexo central e periférico em ratos acordados e uma vez que o ATP se metaboliza rapidamente em ADO, esta teria ação no balanço da resposta quimiorreceptora no RTN. / ATP-mediated purinergic signaling at the level of the rostral ventrolateral medulla (RVLM) contributes to both central and peripheral chemoreceptor control of breathing and blood pressure within the retrotrapezoid nucleus (RTN). However, potential contributions of purinergic signaling in the RTN to cardiorespiratory function in conscious animals has not been tested. We show that in the absence of functional C1 cells, ATP into the RTN increased cardiorespiratory output by a P2-recepor dependent mechanism. We also show that a non-specific P2 receptor blocker (PPADS) reduced the ventilatory response to hypercapnia (7% CO2) and hypoxia (8% O2) in unanesthetized awake rats. Conversely, a specific P2Y1-receptor blocker (MRS2179) into the RTN had no measurable effect on respiratory responses elicited by hypercapnia or hypoxia. Moreover, adenosine (ADO) into the RTN could attenuate the hypercapnia-induced increase in ventilation in vivo and firing rate in RTN neurons in vitro. These results demonstrate that ATP-mediated purinergic signaling contributes to central and peripheral chemoreflex control of breathing in awake rats and ADO could provide a balance between ATP stimulation and its inhibition in RTN during hipercapnia.
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Somatosensory processing by rat medial pontomedullary reticular formation neurones : responses to innocuous and noxious thermal and mechanical stimuliFarham, Craig Jeffrey January 1991 (has links)
This work examines somatosensory processing in "giant" neurones of the medial pontomedullary reticular formation (PMRF) in the rat, with particular emphasis on the response to cutaneous thermal stimuli. Thermal test stimuli were employed as these were deemed to be more precisely quantifiable than other forms of cutaneous stimulation. Activity was recorded from 235 PMRF neurones in 94 female Long Evans rats (270 to 320 g) anaesthetised with urethane (1,25g/kg, i.p.). Rectal temperature was closely controlled at 38 ± 0,5°C. Standard stereotactic and extracellular recording techniques were employed. PMRF giant neurones were identified by their stereotactic location, large, stable spike amplitudes of long duration, responses to cutaneous mechanical stimuli and receptive field properties, and spontaneous discharge characteristics. Ramp, step and sine wave cutaneous thermal stimuli (35-48 °C) were applied to the glabrous skin on the hindpaw by means of a computer-controlled Peltier device. The location of the units was confirmed by subsequent histology. One hundred and eleven neurones were located in nucleus reticularis pontis caudalis (NPC), and 124 in nucleus reticularis gigantocellularis (NGC). Mechanical stimulation excited 188 of 235 (80%) PMRF neurones (ON-m cells), and inhibited 40 (17%, OFF-m cells). Seven cells (3%) had mosaic receptive fields of excitation and inhibition (complex responses, CX-m). Twenty-eight percent of neurones were responsive to both weak and intense stimuli (mixed neurones). The remainder (72%) responded only to intense mechanical stimulation of the skin (high threshold neurones). The (excitatory or inhibitory) response of the mixed neurones to intense stimuli was generally greater than to mild stimuli, Receptive fields ranged in size from restricted (hindlimbs only) to very extensive (covering the entire body surface). Neurones with small receptive fields were almost exclusively of the high threshold type, and tended to be located in NGC, while mixed neurones tended to have larger receptive fields, and were located predominantly in NPC. Some portion of the hind limbs were represented in the receptive fields of all but one of the neurones studied, while the tail and/ or trunk were represented in 77%, and the forelimbs and face in 28% of receptive fields. Most of the cells responding to cutaneous mechanical stimulation had bilateral (usually symmetric) receptive fields. Spontaneous (background) activity occurred in the absence of any deliberate sensory stimulation in 72% of PMRF neurones. The frequency of spontaneous discharge rates ranged from O to 47 spikes/ s. The coefficient of variation of the spontaneous discharge rate of a given neurone was generally less than 20% (range O to 85%). Of the 235 identified mechanosensitive PMRF neurones, 203 (86%) also responded to cutaneous thermal stimulation (43-48 °C) of the ipsilateral hind paw. Eighty percent of these responded with increased discharge rates (ON-t cells), and 20% were inhibited (OFF-t cells). The polarities of response of individual PMRF neurones to mechanical and thermal stimuli, and to repeated ipsilateral and contralateral thermal stimuli, did not differ significantly. Following transient thermal stimulation, spontaneous discharge rates largely returned to pre-stimulus levels. The thresholds of response to slow ramp (0,15°C/s) and stepped (2°C/s) thermal stimuli occurred both in the innocuous and noxious temperature ranges (below and above 42°C, respectively). The threshold temperatures showed large variability to repeated identical thermal stimuli. Despite the poor reproducibility of the threshold responses, the distribution of thresholds to thermal ramp stimuli was consistently bimodal, with peaks occurring at 39 and 43°C. The bimodality persisted even when the ipsilateral and contralateral data were pooled. The modes of these threshold distributions conform to the maximum discharge ranges for warm and noxious cutaneous receptors. Thus, it is likely that thermal input to individual PMRF neurones is derived from both types of receptors. The responses of PMRF neurones to repeated thermal stimuli were stable and reproducible with respect to magnitude and time course. The average (static) and maximum (dynamic) responses to thermal stimuli were generally small: for example, the mean of the average responses to ramp stimuli was 5,9 spikes/s ± 11,0 SD, (range -28 to 40 spikes/s), and the mean of the maximum responses was 9,3 spikes/s ± 16,1 SD, (range -46 to 65 spikes/s). The absolute change in firing rate of individual PMRF neurones, and of the population, increased monotonically as a function of the intensity of stepped cutaneous thermal stimuli in the range 40 to 48 °C. However, their resolution, based on their average and maximum responses, was poor. Incorporating the post-stimulus responses into the comparisons between different stimulus intensities marginally increased the resolution of these neurones. Thus, while the majority of PMRF neurones are able to distinguish innocuous from noxious stimuli, few are capable of encoding stimulus intensity within the noxious range (above 43 °C). The majority (70%) of PMRF neurones responded to sustained thermal stimuli with a slow increase or decrease to a new static discharge rate which was maintained with little or no adaptation. Latency to onset of response to stepped thermal stimuli varied from 1 to 50 seconds, and the time to maximal response between 5-60 seconds. Many PMRF neurones also showed marked after-discharge for periods of up to 5 minutes after removal of the stimulus. The thermal receptive fields of over 90% of PMRF neurones were large, incorporating at least both hindlimbs. The extensive receptive field sizes of individual PMRF neurones provides evidence against them having a role in stimulus location. The large number of PMRF neurones showing multimodal convergence, their small magnitude responses, their slow response times, and their large receptive fields strongly suggest that these neurones are not participating in classical sensory discrimination. Rather, they may function as stimulus detectors or alternatively play a role in associative processes.
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Estudo anatômico do tronco encefálico do macaco Cebus apella / Anatomical study of the brain stem of the Cebus apella monkeyMarques, Karina do Valle 28 March 2005 (has links)
Os macacos Cebus apella são hábeis com suas mãos e capazes de utilizar ferrementas (paus e pedras) para resolver problemas, podendo ser treinados como assistentes de pessoas quadriplégicas. São escassas ainda as informações em neuroanatomia desses primatas, o que motiva a presente pesquisa, cujo propósito é o de enfocar a morfologia externa e interna do tronco encefálico. Foram estudados 20 espécimes do macaco Cebus apella que já estavam fixados com formol a 10%, pertencentes ao acervo de pesquisa do laboratório de Anatomia da Universidade Federal de Uberlândia. Após isolamento do encéfalo e remoção das meninges procedeu ? se ao estudo da superfície externa em 14 peças. A dissecção por técnica usual preservou características morfológicas da superfície e origem aparente de nervos cranianos, que foram identificados. Em análise macroscópica, o tronco encefálico mostra ? se bem desenvolvido destacando ? se ainda em sua face lateral a origem aparente do nervo trigêmeo e os pedúnculos cerebelares médios, bem evidentes. Na face dorsal, em relação ao IV ventrículo identificam ? se os véus medulares superior e inferior, bem como o sulco mediano e a eminência medial. Os pedúnculos cerebelares superior e inferior também são bem destacados. Na medula oblonga são identificados, entre outros, os relevos correspondentes às olivas e às pirâmides, além da fissura mediana anterior e a região da decussação das pirâmides, Nem sempre são nítidos o sulco bulbo ? pontino e o forame cego. Os fascículos grácil e cuneiforme são bastante desenvolvidos bem como os tubérculos correspondentes. Para os estudos das estruturas internas do tronco encefálico, 6 peças, já fixadas, foram congeladas com nitrogênio líquido e cortadas no criostato entre ?27º C e ? 30º C, seguindo ? se a feitura de cortes transversais, em seis níveis (colículos superior e inferior, origem aparente do nervo trigêmeo, porções superior, média e inferior da oliva, e decussação das pirâmides), com espessura de 60 µc, corados segundo o método de Mulligan. As estruturas evidenciáveis são descritas com detalhes comparando ? se os dados obtidos com os da literatura consultada. Os termos anatômicos foram referidos segundo a Terminologia Anatômica Internacional, 1ª edição brasileira ? 2001, Ed. Manole Ltda. / The Cebus apella monkey are able to use their nands and to use tools like woods and stones to solve problems. They can be trained to act as quadriplégicas people?s assistants. The Neuroanatomy information about these primates are little, that?s the reason for this research and its purpose is to accomplish a macroscophial anatomic study of encephalic trunk external and internal surface structure. It was studied Cebus apella monkeu 20 especimens that was already fixed on 10% formaldehyde. These monkeys are from research?s heap of Universidade Federal de Uberlândia. The monkey encephalon were removed using an osteótomo. After removing the meninges, theirs external surface study was started on 14 of them. The dissection using usual technique could preserve the morphological characteristics of the surface and cranial nerve apparent origim, that could be identified too. Under a macroscophical analysis, the encephalic trun looks very elaborated and the trigemini nerve and pedunculus cerebellaris aparent origin, very evident, was stood out in its lateral face. In the dorsal face, in relation to fourth ventricle, the inferior and superior velum medullare could be identified as well as the sulcus medianus and the eminentia medialis. The superior pedunculus cerebellaris, as well as the inferior ones, were very stood out too. On the medulla oblongata were identified, beyond others, the relieves corresponding to the oliva and the decussatio pyramidum region. The sulcus bulbus ? pons and the foramen caecum not always are bright. The fasciculus gracilis, as well as the cuneatus ones, are very elaborated as well as the corresponding tuberculuns.The brain stem internal structure study 6 already fixed pieces were froze with liquid nitrong and cut in criostato, under the temperature between ?27ºC and ? 30º C. After this, transversal cuts were made on 6 levels (superior and inferior nerve trigemini aparent origin, olivas superior, medium and inferior portion and decussatio pyramidum), with the thickness of 60µc, roddy according to the Mulligan method. The evidentiable structures are described in details comparing data obtained from the consulted literature. The anatomic terms were aforesaid according to Terminologia Anatômica Internacional, 1ª edição brasileira ? 2001, Ed. Manole Ltda.
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Alterações na glia observadas em regiões encefálicas envolvidas no controle respiratório em um modelo animal da doença de Parkinson. / Glial changes observed in medullary regions involved in the control of breathing in an animal model of Parkinson\'s disease.Carvalho, Kárin Santana de 16 May 2017 (has links)
A doença de Parkinson (DP) apresenta declínio da capacidade respiratória. Estudo utilizando um modelo de DP induzido pela injeção de 6-hidroxidopamina (6-OHDA) no estriado mostrou redução no número de neurônios envolvidos no controle da respiração. O objetivo desse estudo foi avaliar alterações gliais no núcleo retrotrapezóide (RTN), núcleo do trato solitário, complexo pré-Bötzinger e grupamento respiratório ventrolateral rostral (rVRG) utilizando o mesmo modelo experimental. Em ratos Wistar que receberam a injeção de 6-OHDA bilateralmente no estriado observou-se redução na imunorreatividade para proteína ácida fibrilar glial em astrócitos no rVRG a partir de 30 dias, no RTN a partir de 40 dias e nas demais regiões a partir de 60 dias. Ainda, observamos o aumento da proteína ligante de actina cruzada em microglias na região do RTN aos 30 e 40 dias. Nossos dados sugerem que a redução astrocitária contribui para as alterações respiratórias observadas neste modelo experimental e a presença de neuroinflamação no RTN pode contribuir para a perda celular nesta região. / Parkinson\'s disease (PD) has a decline in respiratory capacity. A study using a model of PD induced by the injection of 6-hydroxydopamine (6-OHDA) in the striatum showed a reduction in the number of neurons involved in the control of breathing. The aim of this study was to evaluate glial changes in the retrotrapezoid nucleus (RTN), nucleus of the solitary tract, pre-Bötzinger complex and rostral ventral respiratory group (rVRG) using the same experimental model. Wistar rats that received bilateral 6-OHDA injection in the striatum present a reduction in immunoreactivity for glial fibrillary acidic protein in astrocytes in the rVRG from 30 days, in the RTN from 40 days and in the other regions from 60 days. We also observed an increase of the Ionized calcium binding adaptor molecule 1 in microglia in the region of the RTN at 30 and 40 days. Our data suggest that astrocytic reduction contributes to the respiratory changes observed in this experimental model and the presence of neuroinflammation in the RTN may contribute to the cellular loss in this region.
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Alterações na glia observadas em regiões encefálicas envolvidas no controle respiratório em um modelo animal da doença de Parkinson. / Glial changes observed in medullary regions involved in the control of breathing in an animal model of Parkinson\'s disease.Kárin Santana de Carvalho 16 May 2017 (has links)
A doença de Parkinson (DP) apresenta declínio da capacidade respiratória. Estudo utilizando um modelo de DP induzido pela injeção de 6-hidroxidopamina (6-OHDA) no estriado mostrou redução no número de neurônios envolvidos no controle da respiração. O objetivo desse estudo foi avaliar alterações gliais no núcleo retrotrapezóide (RTN), núcleo do trato solitário, complexo pré-Bötzinger e grupamento respiratório ventrolateral rostral (rVRG) utilizando o mesmo modelo experimental. Em ratos Wistar que receberam a injeção de 6-OHDA bilateralmente no estriado observou-se redução na imunorreatividade para proteína ácida fibrilar glial em astrócitos no rVRG a partir de 30 dias, no RTN a partir de 40 dias e nas demais regiões a partir de 60 dias. Ainda, observamos o aumento da proteína ligante de actina cruzada em microglias na região do RTN aos 30 e 40 dias. Nossos dados sugerem que a redução astrocitária contribui para as alterações respiratórias observadas neste modelo experimental e a presença de neuroinflamação no RTN pode contribuir para a perda celular nesta região. / Parkinson\'s disease (PD) has a decline in respiratory capacity. A study using a model of PD induced by the injection of 6-hydroxydopamine (6-OHDA) in the striatum showed a reduction in the number of neurons involved in the control of breathing. The aim of this study was to evaluate glial changes in the retrotrapezoid nucleus (RTN), nucleus of the solitary tract, pre-Bötzinger complex and rostral ventral respiratory group (rVRG) using the same experimental model. Wistar rats that received bilateral 6-OHDA injection in the striatum present a reduction in immunoreactivity for glial fibrillary acidic protein in astrocytes in the rVRG from 30 days, in the RTN from 40 days and in the other regions from 60 days. We also observed an increase of the Ionized calcium binding adaptor molecule 1 in microglia in the region of the RTN at 30 and 40 days. Our data suggest that astrocytic reduction contributes to the respiratory changes observed in this experimental model and the presence of neuroinflammation in the RTN may contribute to the cellular loss in this region.
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Estudo anatômico do tronco encefálico do macaco Cebus apella / Anatomical study of the brain stem of the Cebus apella monkeyKarina do Valle Marques 28 March 2005 (has links)
Os macacos Cebus apella são hábeis com suas mãos e capazes de utilizar ferrementas (paus e pedras) para resolver problemas, podendo ser treinados como assistentes de pessoas quadriplégicas. São escassas ainda as informações em neuroanatomia desses primatas, o que motiva a presente pesquisa, cujo propósito é o de enfocar a morfologia externa e interna do tronco encefálico. Foram estudados 20 espécimes do macaco Cebus apella que já estavam fixados com formol a 10%, pertencentes ao acervo de pesquisa do laboratório de Anatomia da Universidade Federal de Uberlândia. Após isolamento do encéfalo e remoção das meninges procedeu ? se ao estudo da superfície externa em 14 peças. A dissecção por técnica usual preservou características morfológicas da superfície e origem aparente de nervos cranianos, que foram identificados. Em análise macroscópica, o tronco encefálico mostra ? se bem desenvolvido destacando ? se ainda em sua face lateral a origem aparente do nervo trigêmeo e os pedúnculos cerebelares médios, bem evidentes. Na face dorsal, em relação ao IV ventrículo identificam ? se os véus medulares superior e inferior, bem como o sulco mediano e a eminência medial. Os pedúnculos cerebelares superior e inferior também são bem destacados. Na medula oblonga são identificados, entre outros, os relevos correspondentes às olivas e às pirâmides, além da fissura mediana anterior e a região da decussação das pirâmides, Nem sempre são nítidos o sulco bulbo ? pontino e o forame cego. Os fascículos grácil e cuneiforme são bastante desenvolvidos bem como os tubérculos correspondentes. Para os estudos das estruturas internas do tronco encefálico, 6 peças, já fixadas, foram congeladas com nitrogênio líquido e cortadas no criostato entre ?27º C e ? 30º C, seguindo ? se a feitura de cortes transversais, em seis níveis (colículos superior e inferior, origem aparente do nervo trigêmeo, porções superior, média e inferior da oliva, e decussação das pirâmides), com espessura de 60 µc, corados segundo o método de Mulligan. As estruturas evidenciáveis são descritas com detalhes comparando ? se os dados obtidos com os da literatura consultada. Os termos anatômicos foram referidos segundo a Terminologia Anatômica Internacional, 1ª edição brasileira ? 2001, Ed. Manole Ltda. / The Cebus apella monkey are able to use their nands and to use tools like woods and stones to solve problems. They can be trained to act as quadriplégicas people?s assistants. The Neuroanatomy information about these primates are little, that?s the reason for this research and its purpose is to accomplish a macroscophial anatomic study of encephalic trunk external and internal surface structure. It was studied Cebus apella monkeu 20 especimens that was already fixed on 10% formaldehyde. These monkeys are from research?s heap of Universidade Federal de Uberlândia. The monkey encephalon were removed using an osteótomo. After removing the meninges, theirs external surface study was started on 14 of them. The dissection using usual technique could preserve the morphological characteristics of the surface and cranial nerve apparent origim, that could be identified too. Under a macroscophical analysis, the encephalic trun looks very elaborated and the trigemini nerve and pedunculus cerebellaris aparent origin, very evident, was stood out in its lateral face. In the dorsal face, in relation to fourth ventricle, the inferior and superior velum medullare could be identified as well as the sulcus medianus and the eminentia medialis. The superior pedunculus cerebellaris, as well as the inferior ones, were very stood out too. On the medulla oblongata were identified, beyond others, the relieves corresponding to the oliva and the decussatio pyramidum region. The sulcus bulbus ? pons and the foramen caecum not always are bright. The fasciculus gracilis, as well as the cuneatus ones, are very elaborated as well as the corresponding tuberculuns.The brain stem internal structure study 6 already fixed pieces were froze with liquid nitrong and cut in criostato, under the temperature between ?27ºC and ? 30º C. After this, transversal cuts were made on 6 levels (superior and inferior nerve trigemini aparent origin, olivas superior, medium and inferior portion and decussatio pyramidum), with the thickness of 60µc, roddy according to the Mulligan method. The evidentiable structures are described in details comparing data obtained from the consulted literature. The anatomic terms were aforesaid according to Terminologia Anatômica Internacional, 1ª edição brasileira ? 2001, Ed. Manole Ltda.
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Mecanismos Cardio-Respiratórios Integrados na Região Bulbar: Participação da Área Pressora Caudal, dos Núcleos do Trato Solitário e do Núcleo Retrotrapezóide / Integrated Cardio-Respiratory Mechanisms within the brainstem: Involvement of the Caudal Pressor Area, the Nucleus of the Solitary Tract and the Retrotrapezopid NucleusTakakura, Ana Carolina Thomaz [UNIFESP] 28 January 2009 (has links) (PDF)
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Publico-086c.pdf: 1776030 bytes, checksum: d420deb32e6a60ac8db7b56dccce3684 (MD5) / Respostas cardiovasculares podem ser integradas em diferentes níveis do sistema nervoso central (SNC). No bulbo existem várias áreas importantes para o controle cardiovascular. Sabe-se que, na superfície ventrolateral do bulbo, existem pelo menos três regiões relevantes ao controle cardiovascular: região rostroventrolateral (RVL), região caudoventrolateral (CVL) e a área pressora caudal (APC). A região RVL tem importante função na manutenção do tônus vasomotor e a região CVL interfere na regulação da pressão arterial (PA) por modular a atividade da região RVL. Além disso, essas duas regiões participam na interação de reflexos cardiovasculares, que incluem a participação de outra estrutura bulbar (Núcleos do Trato Solitário - NTS) como primeira sinapse de fibras aferentes reflexas. A APC foi recentemente descoberta e interfere na regulação da PA por modular a atividade dos neurônios das regiões RVL e CVL. Desse modo, o presente estudo procurou investigar a participação do NTS (região comissural e intermediária) nas respostas produzidas pela estimulação ou inibição da APC. Foram utilizados ratos Holtzman (280-350 g; n = 6-8) anestesiados com uretana (1,2 g/kg, i.v.). Um cateter foi inserido na artéria e veia femoral desses animais para registro de pressão arterial, freqüência cardíaca e injeção de drogas. Em alguns animais, foi registrada a atividade elétrica do nervo esplâncnico e do nervo frênico. Os animais foram adaptados a um aparelho estereotáxico e submetidos a uma craniotomia parcial para injeções de drogas. Num primeiro grupo experimental, os animais foram submetidos a lesão fictícia ou eletrolítica (1 mA x 10s) do NTS comissural (NTScom). A injeção unilateral do aminoácido excitatório L-glutamato (1 mM - 100 nl) na APC produziu resposta pressora (D = +28 ± 3 mmHg, vs. controle: D = +2 ± 2 mmHg, p<0,05) e taquicárdica (D = +20 ± 1 bpm, vs. controle: D = +3 ± 3 bpm, p<0,05). As respostas pressora e taquicárdica promovidas pela injeção unilateral de L-glutamato na APC foram abolidas 10 (D = +5 ± 7 mmHg e D = +4 ± 3 bpm, respectivamente), 30 (D = +7 ± 3 mmHg e D = +6 ± 3 bpm, respectivamente) e 60 minutos (D = +11 ± 3 mmHg e D = +5 ± 1 bpm, respectivamente) após a lesão eletrolítica do NTScom. Da mesma maneira, no grupo de animais que recebeu a injeção do agonista GABA-A muscimol no NTScom, as respostas pressora e taquicárdica promovidas pela injeção unilateral de L-glutamato na APC foram abolidas 10 (D = +5 ± 5 mmHg e D = +5 ± 3 bpm, respectivamente), 30 (D = +8 ± 5 mmHg e D = +7 ± 2 bpm, respectivamente) e 60 minutos (D = +11 ± 6 mmHg e D = +5 ± 2 bpm, respectivamente) após a injeção de muscimol. Por outro lado, as respostas pressora e taquicárdica promovidas pela injeção unilateral de L-glutamato na APC não foram modificadas 10 (D = +11 ± 2 mmHg e D = +30 ± 12 bpm, respectivamente), 30 (D = +27 ± 4 mmHg e D = +32 ± 15 bpm, respectivamente) e 60 minutos (D = +24 ± 7 mmHg e D = +24 ± 8 bpm, respectivamente) após as injeções bilaterais de muscimol no NTS intermediário. Dessa forma, as repostas de estimulação da APC parecem depender da integridade dos neurônios localizados no NTScom. Sendo assim, será que a integridade da APC é importante para as respostas de estimulação do NTScom? Apenas as injeções bilaterais do agonista GABAérgico muscimol (2 mM - 100 nl) na APC promoveu queda de PAM (D = -18 ± 6 mmHg, p<0,05) e FC (D = - 21 ± 8 bpm, p<0,05). As respostas depressora e bradicárdica promovidas pela injeção unilateral de L-glutamato no NTScom foram abolidas 10 (D = -5 ± 3 mmHg e D = +8 ± 9 bpm, respectivamente), 30 (D = -5 ± 2 mmHg e D = +7 ± 4 bpm, respectivamente) e 60 minutos (D = -6 ± 4 mmHg e D = +10 ± 6 bpm, respectivamente) após as injeções bilaterais de muscimol na APC. Da mesma maneira do ocorrido com as respostas de estimulação da APC, as respostas depressora e bradicárdica promovidas pela injeção unilateral de L-glutamato no NTS intermediário não foram modificadas 10 (D = -41 ± 9 mmHg e D = -67 ± 26 bpm, respectivamente), 30 (D = -31 ± 12 mmHg e D = -23 ± 4 bpm, respectivamente) e 60 minutos (D = -32 ± 8 mmHg e D = -21 ± 3 bpm, respectivamente) após as injeções bilaterais de muscimol na APC. Nossa próxima questão era saber se a inibição da APC promoveria alterações nos reflexos cardiovasculares. A curva do barorreflexo foi construída com injeções endovenosas de uma dose depressora de nitroprussiato de sódio (30 mg/kg) e com a inflação de um balão na aorta torácica. A relação de atividade simpática e PAM (curva do barorreflexo) não revelou nenhum efeito significativo alterado após as injeções bilaterais de muscimol na APC. Após a inibição da APC o barorreflexo continuou operando nos mesmos valores de PAM50 quando comparado ao grupo controle. Os valores de platô superior e inferior, bem como o ganho também não tiveram diferença quando comparados com o grupo controle. A ativação do quimiorreflexo foi realizada com injeções endovenosas de cianeto de sódio (NaCN – 50 mg/kg). A ativação do quimiorreflexo com NaCN produziu resposta pressora (D = +16 ± 4 mmHg, vs. salina D = +2 ± 2 mmHg, p<0,05), aumento da atividade simpática do nervo esplâncnico (D = +236 ± 11% em relação ao controle, p<0,01) e aumento da atividade do nervo frênico. Muscimol injetado bilateralmente na APC promoveu uma redução da resposta pressora (D = +9 ± 2 mmHg, vs. salina D = +18 ± 4 mmHg, p<0,05), da resposta simpatoexcitatória (D = +88 ± 18%, vs. salina D = +217 ± 33 %, p<0,01) e do aumento da atividade do nervo frênico produzido pela ativação dos quimiorreceptores periféricos com NaCN. As aferências dos quimiorreceptores periféricos terminam predominantemente no NTScom. Para corroborar a interação existente entre os neurônios do NTScom e a APC, o próximo experimento foi realizado para testar se os neurônios localizados no NTS ou na APC, ativados por hipóxia, são predominantemente glutamatérgicos ou gabaérgicos. Os animais foram expostos a situações de hipóxia (8% O2, n = 5) ou normóxia (ar ambiente, n = 4). A imunorretividade para a proteína Fos foi utilizada para identificação dos neurônios ativados por hipóxia. Os neurônios glutamatérgicos ou gabaérgicos foram definidos devido à expressão, via hibridzação in situ, de VGLUT2 mRNA ou GAD67 mRNA, respectivamente. A maioria dos neurônios que expressam Fos após hipóxia são glutamatérgicos na região do NTScom (64 ± 13%) e NTS intermediário (41 ± 9%). Na região da APC, dos neurônios que expressam Fos após hipóxia, somente uma pequena parcela (26 ± 9%) são glutamatérgicos. Apenas uma pequena quantidade de neurônios que expressam Fos também expressam GAD67 mRNA na região do NTScom (7 ± 4%), na região do NTS intermediário (16 ± 9%) e na APC (5 ± 3%). A imunorreatividade para a proteína Fos foi praticamente inexistente nos animais que foram expostos ao ar ambiente. Diante disso, os nossos resultados são os primeiros a mostrar a existência da interação entre a região comissural do NTS e a APC no que diz respeito ao controle cardiovascular. Sabe-se que a região do NTScom contém neurônios de segunda ordem que recebem as aferências dos quimiorreceptores periféricos e que as respostas de estimulação do NTScom parecem depender da integridade da APC. Logo, os resultados do presente estudo também mostraram que a inibição da APC foi capaz de reduzir as respostas simpatoexcitatórias e respiratórias após a estimulação dos quimiorreceptores periféricos. Por outro lado, a inibição da APC não foi capaz de reduzir as respostas de ativação do barorreflexo. / Cardiovascular responses are integrated at different levels of the central nervous system (CNS). In the brainstem, there are different areas related to the cardiovascular control. It is well known that in the ventrolateral medulla there are, at least, three important regions involved with cardiovascular control: the rostral ventrolateral medulla (RVLM), the caudal ventrolateral medulla (CVLM) and the caudal pressor area (CPA). The RVLM is the main source of excitatory input to sympathetic preganglionic neurons playing a crucial role for tonic and reflex control of the cardiovascular system. The caudal ventrolateral medulla (CVLM) contains the GABAergic sympathoinhibitory neurons that project to RVLM. Besides these two regions, the NTS is the site of the first synapse of the viscerosensory afferent fibers in the brainstem including those related to cardiovascular and respiratory afferents. Besides the circuitry described above, more recent studies have also shown a new site, the CPA, located in the caudal end of the ventrolateral medulla, which provides excitatory signals that affect sympathetic activity to the cardiovascular system in rats. Therefore, the aim of the present study was to investigate the involvement of the NTS (commissural and/or intermediate regions) on the stimulation or inhibition of the CPA. Male Holtzman rats (280-350 g; n = 6-8) anesthetized with urethane (1.2 g/kg, i.v.) were used. Unilateral microinjection of L-glutamate (1 mM - 100 nL) into the CPA resulted in an increase in mean arterial pressure (MAP, D = +28 ± 3 mmHg, vs. controle: D = +2 ± 2 mmHg, p<0.05) and heart rate (HR, D = +20 ± 1 bpm, vs. controle: D = +3 ± 3 bpm, p<0.05). Electrolytic lesions of the commissural NTS (commNTS) (1 mA x 10s) abolished the pressor and tachycardic responses at 10 (D = +5 ± 7 mmHg and D = +4 ± 3 bpm), 30 (D = +7 ± 3 mmHg and D = +6 ± 3 bpm) and 60 minutes (D = +11 ± 3 mmHg and D = +5 ± 1 bpm) produced by injection of L-glutamate into the CPA. In the same way, muscimol (GABA-A receptor agonist) (2 mM - 60 nL) injected into the commNTS abolished the pressor and tachycardic responses at 10 (D = +5 ± 5 mmHg and D = +5 ± 3 bpm), 30 (D = +8 ± 5 mmHg and D = +7 ± 2 bpm) and 60 minutes (D = +11 ± 6 mmHg and D = +5 ± 2 bpm) produced by injection of L-glutamate into the CPA. On the other hand, the injection of Lglutamate unilaterally into the CPA 10, 30 and 60 minutes after the injection of muscimol into the intermediate NTS produced no change on MAP increase (D = +11± 2, +27 ± 4 and +24 ± 7 mmHg, respectively) and HR (D = +30 ± 12, +32 ± 15 and +24 ± 8 bpm, respectively). It seems that the activation of the CPA depends on the integrity of the commNTS neurons. Does the CPA integrity depend on NTS activation? Bilateral injection of muscimol (GABA-A agonist, 2 mM – 60 nl) into the CPA produced a decrease in MAP (D = -18 ± 2 mmHg, p<0.05) and HR (D = -21 ± 6 bpm, p<0.05). The depressor and bradycardia responses produced by L-glutamate into the commNTS was abolished at 10 (D = -5 ± 3 mmHg and D = +8 ± 9 bpm), 30 (D = -5 ± 2 mmHg and D = +7 ± 4 bpm) and 60 minutes (D = -6 ± 4 mmHg and D = +10 ± 6 bpm) after muscimol injection into the CPA. On the other hand, the depressor and bradicardic responses produced by L-glutamate into the intermediate NTS was not changed at 10 (D = -41 ± 9 mmHg and D = -67 ± 26 bpm), 30 (D = -31 ± 12 mmHg and D = -23 ± 4 bpm) and 60 minutes (D = -32 ± 8 mmHg and D = -21 ± 3 bpm) after muscimol injection into the CPA. Our next question was to know if the inhibition of the CPA will change the cardiovascular reflexes. Baroreflex curves were generated by lowering MAP with sodium nitroprusside (30 mg/kg) and increasing MAP by constricting an abdominal aortic snare. The relationship between MAP and sympathetic nerve discharge (SND) (baroreflex curves) revealed no change after muscimol injection into the CPA. After inhibition of the CPA, the baroreflex operated around a comparable MAP (MAP50) in all groups of rats. The lower and upper plateau, as well the baroreflex range was not different from the control group. The chemoreflex was actvated by i.v. injections of sodium cyanide (NaCN – 50 mg/kg). The activation of chemoreflex with NaCN produced a rise in MAP (D = +16 ± 4 mmHg, vs. saline D = +2 ± 2 mmHg, p<0.05), an increase in SND (D = +236 ± 11% of resting, p<0.01) and an increase in phrenic nerve discharge (PND). Muscimol injection into the CPA reduced the pressor (D = +9 ± 2 mmHg, vs. saline D = +18 ± 4 mmHg, p<0.05), sympathoexcitatory (D = +88 ± 18%, vs. saline D = +217 ± 33 %, p<0.01) and the increase in PND produced by chemoreflex activation with NaCN. Carotid chemoreceptor afferents terminate predominantly in commNTS. The next experiment was designed to test and to confirm the interaction between the commNTS and the CPA neurons by anatomical approaches. We tested whether glutamatergic or GABAergic. The rats were exposed to hypoxia (8% O2, n = 5) or to normal air (n = 4). Fos-immunoreactivity was used to identify commNTS neurons that were activated by hypoxia. NTS or CPA neurons were classified as glutamatergic or GABAergic based on whether they contained VGLUT2 mRNA or GAD67 mRNA. The vast majority of the commNTS (64 ± 13%), intermediate NTS (41 ± 9%) and CPA (26 ± 9%) neurons that were immunoreactive for Fos contained VGLUT2 mRNA, whereas only a small proportion of the same class of neurons contained GAD-67 mRNA (commNTS: 7 ± 4%; intermediate NTS: 16 ± 9% and CPA: 5 ± 3%). Therefore, our results are the first to show the existence of an interaction between the commNTS and CPA in terms of cardiovascular control. However we know that commNTS neurons receive inputs from peripheral chemoreceptors and the stimulation of commNTS depends on CPA integrity. These data suggest that CPA contributes to resting MAP, sSND and to the sympathoexcitatory effect during stimulation of peripheral chemoreceptors. / TEDE / BV UNIFESP: Teses e dissertações
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The Human Y chromosome and its role in the developing male nervous systemJohansson, Martin M. January 2015 (has links)
Recent research demonstrated that besides a role in sex determination and male fertility, the Y chromosome is involved in additional functions including prostate cancer, sex-specific effects on the brain and behaviour, graft-versus-host disease, nociception, aggression and autoimmune diseases. The results presented in this thesis include an analysis of sex-biased genes encoded on the X and Y chromosomes of rodents. Expression data from six different somatic tissues was analyzed and we found that the X chromosome is enriched in female biased genes and depleted of male biased ones. The second study described copy number variation (CNV) patterns in a world-wide collection of human Y chromosome samples. Contrary to expectations, duplications and not deletions were the most frequent variations. We also discovered novel CNV patterns of which some were significantly overrepresented in specific haplogroups. A substantial part of the thesis focuses on analysis of spatial expression of two Y-encoded brain-specific genes, namely PCDH11Y and NLGN4Y. The perhaps most surprising discovery was the observation that X and Y transcripts of both gene pairs are mostly expressed in different cells in human spinal cord and medulla oblongata. Also, we detected spatial expression differences for the PCDH11X gene in spinal cord. The main focus of the spatial investigations was to uncover genetically coded sexual differences in expression during early development of human central nervous system (CNS). Also, investigations of the expression profiles for 13 X and Y homolog gene pairs in human CNS, adult brain, testes and still-born chimpanzee brain samples were included. Contrary to previous studies, we found only three X-encoded genes from the 13 X/Y homologous gene pairs studied that exhibit female-bias. We also describe six novel non-coding RNAs encoded in the human MSY, some of which are polyadenylated and with conserved expression in chimpanzee brain. The description of dimorphic cellular expression patterns of X- and Y-linked genes should boost the interest in the human specific gene PCDH11Y, and draw attention to other Y-encoded genes expressed in the brain during development. This may help to elucidate the role of the Y chromosome in sex differences during early CNS development in humans. / <p>chinese, finnish, norwegian, schizophrenia, bipolar, bipolar disorder, msy, male specific region Y, PAR1, PAR2, pseudoautosomal, male-biased, female-biased, male biased, female biased, ashkenazi population, structure, variants, YHRD, Elena Jazin, Björn Reinius, Per Ahlberg, brain, hjärna, CNS, central nervous system, IR2, inverted repeat 2, isodicentric, genetics, genetik, padlock, rolling circle, amplification, PCR, sY1191, sY1291, STS, DDX3Y, DAZ, AZFa, AZFb, AZFc, AZF, Repping, haplogroup J, rearrangements, DE-M145, I-M170, E-M96, Q-M242, R-M207, O-M175, G-M201, D-M174, C-M130, NO-M214, N-M231, poland</p>
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