Our purpose was to identify and localize intrinsic cardiac ganglia innervating distinct regions of the heart using postmortem tracing of nerve projections with DiI, a method not previously used to study the intrinsic cardiac nervous system. We also investigated the possibility of collateral innervation of myocardium and intrinsic ganglia. In isolated paraformaldehyde-fixed guinea pig hearts, crystals of DiI (1,1′- dioctadecyl-3,3,3′,3′-tetramethylin-docarbocyanine perchlorate) were inserted into the posterior ventricular myocardium below the atrioventricular groove, the right atrium, or the left ventricular septum. Hearts were placed in the dark at 37°C for 2-14 weeks to allow DiI diffusion within neuronal membranes. Labeled neurons were observed in intracardiac ganglia after at least 4 weeks of dye exposure. Labeling was restricted to the inferior-most ganglia (those near the atrioventricular groove) when DiI was inserted into the posterior ventricular myocardium and to ganglia near the sinus node after right atrial DiI placement. Application of DiI to the left ventricular septum resulted in neuron labeling in ganglia primarily in the interatrial septum near the atrioventricular node. After 8 weeks, DiI-labeled nerve fibers and varicosities were seen surrounding unlabeled neurons in some ganglia, suggesting that axons terminating in or passing through the DiI application site in posterior ventricular tissue had collateral branches innervating these ganglia. These results indicate that intrinsic innervation of major cardiac subdivisions is accomplished by regionally segregated cardiac ganglia. Also, tracing with DiI has provided evidence for collateral nerve projections that could be the substrate for novel intracardiac regulatory circuits.

Résumé : Introduction : Parmi les structures anatomiques impliqués dans la statique pelvienne, la continence urinaire et anale, le muscle élévateur de l’anus (MEA), le sphincter urétral (SU) et son innervation ont un rôle déterminant. Au cours de la grossesse, de l’accouchement par voie vaginale, de la chirurgie radicale pelvienne, des lésions des muscles du plancher pelvien ou de son innervation peuvent survenir. Ces lésions sont à l’origine de dysfonctions du plancher pelvien telle que le prolapsus uro-génital ou l’incontinence urinaire. Une meilleure connaissance de l’anatomie musculaire et nerveuse pelvi-périnéale est nécessaire pour diminuer la survenue et traiter ces troubles fonctionnels. Classiquement les muscles du plancher pelvien sont décrits comme entièrement striés sous contrôle somatique (nerf du MEA et/ou nerf pudendal (NP)). La dissection Anatomique Assisté par Ordinateur (DAAO) en utilisant des marqueurs nerveux et musculaires spécifiques peut aujourd’hui compléter les données établies par la dissection conventionnelle classique de sujets anatomiques.Objectif : L’objectif était de décrire l’innervation (origine, topographie, trajet, rapports, fonction) et la structure musculaire du MEA, de décrire l’innervation (origine, topographie, trajet, rapports, fonction) du sphincter urétral pour mettre en perspective les implications potentielles dans les dysfonctions du plancher pelvien.Méthodes : Nous avons étudié 9 fœtus humains (6 féminins et 3 masculins). Des coupes histologiques sériées de 5 µm d’épaisseur ont été effectuées dans les régions pelviennes de 7 fœtus âgés de 18 à 40 semaines de gestation. Pour chaque niveau de coupe, des lames ont été colorées puis traitées en immunohistochimie pour détecter : l’ensemble des fibres nerveuses (anticorps anti-protéine S100), les fibres autonomes cholinergiques (anti-VAChT), les fibres autonomes adrénergiques (anti-TH), les fibres autonomes nitrergiques (anti-nNOS), les fibres somatiques (anti-PMP 22), les fibres sensorielles (anti-CGRP), les fibres musculaires lisses (anti-SMA) et les fibres musculaires striées (anti-MYOG). Les coupes ont ensuite été numérisées par un scanner de haute résolution optique et les images ont été reconstruites en 3D avec le logiciel Winsurf®. Un fœtus additionnel a été entièrement destiné à réaliser de la microscopie électronique afin de confirmer nos résultats au niveau architectural musculaire.Résultats : Nous avons observé une innervation à la fois autonome (plexus hypogastrique inférieur (PHI)) et somatique (nerf du MEA et NP) du MEA. Nous avons individualisé des zones de cellules musculaires lisses au sein du plancher pelvien notamment de la partie médiane sous contrôle autonome (PHI) que nous nommons « compartiment médian musculaire lisse ».Nous avons systématisé le plancher pelvien musculaire en une zone médiale lisse sous contrôle nerveux autonome réalisant une interface entre les viscères pelviens, et une zone musculaire striée latérale sous contrôle nerveux somatique.Enfin, nous avons mis en évidence une double innervation à la fois somatique et autonome du sphincter urétral.Conclusion : La DAAO a permis de mettre en évidence une innervation pelvi-périnéale complexe avec l’intrication du système nerveux somatique et autonome. La fonction du contingent musculaire lisse pelvien reste à préciser. / Abstract : Levator ani muscle (LAM), urethral sphincter (US), and their innervation play a major role in pelvic status, urinary continence and anal continence. During pregnancy, vaginal delivery, pelvic radical surgery, damage of pelvic floor muscles or of their innervation may occur. These lesions are responsible for pelvic floor dysfunction such as urogenital prolapse or urinary incontinence. Better knowledge of pelvi-perineal muscle and nerve anatomy is needed to reduce the occurrence and treat these pelvic floor dysfunctions. Classically pelvic floor muscles are described as entirely striated under somatic control (levator ani nerve (LAN) and /or pudendal nerve (PN)). Computer Assisted Anatomical Dissection (CAAD) using specific nerve and muscle markers can nowadays complete data established by conventional dissection of anatomical subjects.The objective was to describe the innervation and the muscular structure of the levator ani muscle, to describe nerve supply of the urethral sphincter in order to better understand pelvic floor dysfunction.Methods : We studied nine human fetuses (6 female and 3 male). Serial histological sections of 5 μm of thickness were performed in the lumbar and pelvic regions of seven human fetuses aged 18 to 40 weeks of gestation. For each level, slides were stained and then treated in immunohistochemistry to detect: general nerve fibers (anti-protein S100), autonomic cholinergic fibers (anti-VAChT), autonomic adrenergic fibers (anti-tyrosine hydroxylase), autonomic nitrergic fibers (anti-nNOS), somatic nerve fibers (anti-peripheral myelin protein 22), sensory fibers (anti-CGRP), smooth muscle fibers (anti-SMA) and striated muscle fibers (anti-MYOG). The slides were then digitized by a high-resolution optical scanner and the images were reconstructed in 3D using the Winsurf® software. One fetus was entirely reserved for electronic microscopy in order to confirm our results.Results: We observed that LAM innervation is supplied by both autonomic (inferior hypogastric plexus (IHP)) and somatic (LAN and PN). We have individualized areas of smooth muscle cells in the pelvic floor, particularly the median part controlled by the autonomic system that we named “smooth muscle medial compartment”.We systematized within the pelvic floor muscle a smooth medial part under an autonomic nervous control as interface between the pelvic viscera and a lateral striated muscle part under somatic nervous control.Finally, we have demonstrated a dual innervation both somatic and autonomic sphincter urethral.Conclusion : CAAD has demonstrated complex pelvic-perineal innervation with the interaction of the somatic and autonomic nervous system. The function of the pelvic smooth muscle contingent remains to be clarified.

The origin of cardiac preganglionic neurons in the rat was investigated using the retrograde transport of horseradish peroxidase (HRP). A single injection of HRP was made into the right myocardium in either a sinoatrial or mid-ventricular location. Labeled cells were found in the mid- and lower medulla primarily in and around the nucleus ambiguus (NA) 600-1800 μm above the obex. The dorsal motor nucleus of X (DMN) was sparsely labeled and a few cells were found in an intermediate zone near the level of the obex. Labeling was bilateral with slightly heavier labeling found ipsilateral to the injection site than contralateral to it. Following a unilateral vagotomy, labeled cells were only found ipsilateral to the intact vagus. Atrial and midventricular injections yielded similar results. Occasionally only 1- cells in the NA were labeled per section. Inspection of serial sections revealed that in these sparsely labeled rats, the HRP was often in the same location within the NA forming a column of cells within the nucleus. The columns sometimes extended at least 240 μm in the rostral-caudal direction. The columnar organization was most apparent in rats with few labeled cells presumably because it was obscured in nuclei that were heavily labeled. In a second group of rats, the right vagus was cut at the cervical level and dipped in HRP to determine the extent of the NA and DMN in rats. In these animals, heavier labeling was found in the DMN than in the NA. Cells in the DMN were filled from the upper spinal cord to its most rostral extent 1200 μm above the obex. Thus, although the DMN and NA send projections in the vagus nerve, those axons terminating in the myocardium primarily originate in the NA.

L'objectif du présent projet était d'optimiser le modèle de peau reconstruite par génie tissulaire déjà mis au point par notre équipe afin qu'il stimule le processus de régénération nerveuse après greffe pour améliorer la récupération tactile des grands brûlés. Nous avons utilisé des follicules pileux murins et des neurones sensoriels extraits des ganglions de la racine dorsale de f?tus de souris et avons posé comme hypothèse que les poils avaient une influence positive sur la migration axonale. Une élongation vigoureuse des neurites a été détectée à l'intérieur du derme reconstruit. En présence de cellules épithéliales, il y a eu une migration préférentielle des axones vers les follicules pileux immatures implantés dans le modèle, migration qui a été suivie d'une association étroite des axones et des follicules pileux. Nous avons donc développé avec succès un modèle nous permettant d'étudier les effets de l'épiderme et des follicules pileux sur la croissance des nerfs.

No description available.

Análise proteômica das glândulas de veneno e acessória durante o ciclo de produção de veneno e a influência da inervação noradrenérgica na síntese de proteínas foi verificada. Nossos dados mostram que a síntese de proteínas envolvidas no processo de síntese e secreção de proteínas está aumentada nos estágios ativados da glândula de veneno e que a estimulação da inervação noradrenérgica regula a síntese de algumas proteínas importantes para estes processos. Mostramos pela primeira vez, a presença de várias toxinas no estágio quiescente da glândula e que a produção e secreção de toxinas ocorrem de maneira não sincronizada. Na glândula acessória verificamos uma modulação na síntese de algumas proteínas da glândula após a extração de veneno. Mostramos também a presença de várias toxinas e de inibidores de enzimas do veneno nesta glândula. A composição proteômica das glândulas do aparelho glandular de veneno de serpente contribuirá para um melhor conhecimento dos mecanismos de produção e secreção de veneno e para os estudos das toxinas e suas diversidades. / Proteomic analysis of venom gland and accessory gland during venom production cycle and the influence of noradrenergic innervation on protein synthesis of venom gland were verified. Our data show that the synthesis of proteins involved in the process of synthesis and secretion of proteins is increased in activated stages of venom gland and the sympathetic outflow regulates the synthesis of some proteins that is important to this process. For the first time we showed that many toxins are present in quiescent stage of venom gland and the production and secretion of toxins is not synchronized. Regarding the accessory gland, we verify that the syntheses of some protein of gland were regulated after venom extraction. We also showed the presence of some toxins of the venom and enzyme inhibitors in this gland. The proteomic composition of snake venom gland apparatus will contribute to better understand the mechanisms involved in venom gland activation and consequently, venom production. These data will also contribute to the studies on snake toxins and their diversities.

In this study, the origin and development of the intrinsic innervation in the foetal mouse lung is described and experimental evidence is provided to support the involvement of glial cell line-derived neurotrophic factor (GDNF) in the guidance of nerves and neuronal precursors in the developing lung. Antibodies were used to stain for neuronal precursors, neurones, nerve fibres, primordial epithelium and smooth muscle. These structures were revealed in whole mounts of foetal mouse lungs by immunofluorescence and confocal microscopy, and their spatial and temporal distribution was mapped from the onset of lung development and through the pseudoglandular period. The results showed that neuronal precursors, positive for neural crest cell markers, were present in the vagal tract of the foregut at embryonic day 10 (E10), the time of the evagination of the lung buds. These neural crest-derived cells (NCC) migrated into the lung at E11, along nerve processes directed from the vagus to the smooth musclecovered trachea and emerging lobar bronchi. During E11-E14, a network of nerves and ganglia became established along the dorsal trachea, and large ganglia formed a plexus at the ventral hilum. Nerve trunks issued from these ganglia, travelled along the smooth muscle-covered bronchi, providing a pathway for migrating NCC. To investigate the role of GDNF in the innervation of the lung, an in vitro model of left lung lobes was established. Lung growth and tubule branching was comparable to that in vivo, and neural tissue and smooth muscle continued to grow and thrive. A significant increase in nerve growth occurred when explants were cultured with GDNF compared to controls. Nerves extended, and NCC migrated towards GDNF-impregnated beads suggesting that GDNF may be the molecule guiding nerve fibres and NCC in the lung. The migrating NCC were negative for GDNF-family receptor α1 (GFRα1) during their migration into the lung while the nerves were positive. Since GDNF needs to be associated with its binding receptor, GFRα1, for cellular signalling, GDNF may induce the migration of the NCC if they migrate along the GFRα1-positive nerve fibres. It is concluded that neural tissue and smooth muscle become integral components of the lung shortly after the onset of lung development. The results show that the migration of neural crest-derived cells into the lung and the establishment of the innervation requires coordinated cross-talk between NCC, nerves and smooth muscle throughout development.

Non-human primates have constituted an important group among animals subjected to various studies, such as ethological and paleontological studies who have revealed changes in anatomical structures linked to the evolution of primates and the detailed knowledge on their anatomy may represent an important factor for their preservation and protection. Aimed to identify the distribution of the femoral, saphenous, obturator and sciatic nerve of the pelvic limb of Cebus libidinosus (C.l.). In general, the enervation model of the four primates, men, baboons, chimpanzees and C.l. is identical, but in specific terms, the differences in enervations indicate evolution divergence closer to the branch of baboons in the evolutionary tree, and these data corroborate other comparative studies in relation to the same primates to vessels, muscles and nerves. In conclusion, the sciatic nerve sends branches to the biceps femoris muscle, semitendinous, semimebranosus and accessory and to the ischiofemoral muscle, in the distal third, this nerve is divided into tibial, sural cutaneous medial and common fibular nerves; the femoral nerve sends branches to the sartorius muscle, vastus intermedius muscle, vastus medialis muscle, to the vastus lateralis muscle and branches to the rectus femoris muscle and sends the saphenous nerve. The obturator nerve provides branches to the pectineus muscles, adductor brevis muscles, adductor magnus muscles, adductor longus muscle and glacilis muscle. / Os primatas não-humanos têm constituído um grupo importante dentre os animais submetidos a diversos estudos, tais como etológicos e paleontológicos que têm revelado mudanças nas estruturas anatômicas associadas à evolução dos primatas e o conhecimento detalhado sobre a sua anatomia pode representar um fator importante para a sua preservação e proteção. Objetivou-se verificar a distribuição dos nervos femoral, safeno, obturatório e isquiático do membro pélvico de Cebus libidinosus (C.l.). Em geral, o modelo de inervação dos quatro primatas, homens, babuínos, chipanzés e C.l. é semelhante, mas em casos específicos, as diferenças nas inervações indicam divergência evolucionária mais próxima ao grupo dos babuínos na árvore evolutiva, e esses dados corroboram outros estudos comparativos em relação aos mesmos primatas quanto aos vasos, músculos e nervos. Em conclusão, o nervo isquiático emite ramos para os músculos bíceps femoral, semitendíneo, semimembranáceo próprio, semimembranáceo acessório e isquiofemoral e no terço distal da coxa, este nervo está dividido em nervos tibial, cutâneo sural medial e fibular comum; o nervo femoral envia ramos para os músculos sartório, vastos medial, intermédio e lateral, retofemoral e emite o nervo safeno e o nervo obturatório fornece ramos para os músculos pectíneo, adutores curto, longo e magno, grácil. / Mestre em Ciências Veterinárias

Made available in DSpace on 2014-12-17T15:36:58Z (GMT). No. of bitstreams: 1 RovenaCGJE_DISSERT.pdf: 2142357 bytes, checksum: bbf33b27b7a838600336db16673db325 (MD5) Previous issue date: 2010-04-23 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior / 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 / O complexo parabraquial (PB) ? uma regi?o do tronco encef?lico respons?vel pelo processamento e transmiss?o de informa??es fisiol?gicas essenciais para a sobreviv?ncia dos organismos. Essa regi?o ? subdividida em aproximadamente nove regi?es, considerando caracter?sticas morfol?gicas, citoarquitet?nicas e funcionais. Seus neur?nios possuem uma ampla rede de conex?es com as demais regi?es do sistema nervoso. O objetivo deste trabalho foi mapear a proje??o retiniana para o PB e fazer uma caracteriza??o citoarquitet?nica e neuroqu?mica desta regi?o no Callithrix jacchus (sag?i), um primata do Novo Mundo. As proje??es retinianas foram mapeadas por transporte anter?grado da subunidade B da toxina col?rica (CTb). A citoarquitetura foi descrita atrav?s do m?todo de Nissl e a caracteriza??o neuroqu?mica foi feita atrav?s de t?cnicas imunoistoqu?micas para alguns neurotransmissores e subst?ncias neuroativas presentes neste centro neural. No PB do sagui, nas sec??es coronais coradas pelo m?todo de Nissl, foi poss?vel encontrar um padr?o similar ao que ? evidenciado em outras esp?cies animais. A imunorreatividade contra CTb foi encontrada em fibras terminais do PBMv, caracterizando desta forma uma inerva??o retiniana nessa ?rea. A t?cnica imunoistoqu?micas revelou que o PB cont?m c?lulas, fibras e/ou terminais imunorreativos a prote?na nuclear neuronal, colina acetiltransferase, ?xido n?trico sintetase, serotonina, encefalina, subst?ncia P, prote?nas ligantes de c?lcio (calbindina, calretinina e parvalbumina), e a prote?na ac?dica fibrilar glial. A partir de t?cnicas histoqu?micas verificou-se c?lulas e fibras reativas a NADPH-diaforase. Cada uma dessas subst?ncias apresentou um padr?o caracter?stico de marca??o no PB e algumas serviram como marcadores espec?ficos de subregi?es

Análise proteômica das glândulas de veneno e acessória durante o ciclo de produção de veneno e a influência da inervação noradrenérgica na síntese de proteínas foi verificada. Nossos dados mostram que a síntese de proteínas envolvidas no processo de síntese e secreção de proteínas está aumentada nos estágios ativados da glândula de veneno e que a estimulação da inervação noradrenérgica regula a síntese de algumas proteínas importantes para estes processos. Mostramos pela primeira vez, a presença de várias toxinas no estágio quiescente da glândula e que a produção e secreção de toxinas ocorrem de maneira não sincronizada. Na glândula acessória verificamos uma modulação na síntese de algumas proteínas da glândula após a extração de veneno. Mostramos também a presença de várias toxinas e de inibidores de enzimas do veneno nesta glândula. A composição proteômica das glândulas do aparelho glandular de veneno de serpente contribuirá para um melhor conhecimento dos mecanismos de produção e secreção de veneno e para os estudos das toxinas e suas diversidades. / Proteomic analysis of venom gland and accessory gland during venom production cycle and the influence of noradrenergic innervation on protein synthesis of venom gland were verified. Our data show that the synthesis of proteins involved in the process of synthesis and secretion of proteins is increased in activated stages of venom gland and the sympathetic outflow regulates the synthesis of some proteins that is important to this process. For the first time we showed that many toxins are present in quiescent stage of venom gland and the production and secretion of toxins is not synchronized. Regarding the accessory gland, we verify that the syntheses of some protein of gland were regulated after venom extraction. We also showed the presence of some toxins of the venom and enzyme inhibitors in this gland. The proteomic composition of snake venom gland apparatus will contribute to better understand the mechanisms involved in venom gland activation and consequently, venom production. These data will also contribute to the studies on snake toxins and their diversities.