Urethroplasty using a bovine pericardium graft: an experimental study using normal urethras from dogs

The use of bovine pericardium as a urethral patch to substitute a ventral segment of canine urethras was studied. Healing, epithelial growth, urethral permeability, fistulas, and calcification were analyzed. Thirty male mongrel dogs of medium and large size underwent resection of a ventral segment of the medial urethra measuring 2.0 x 0.5 cm, which was replaced with a bovine pericardium graft, treated with buffered glutaraldehyde and preserved in formaldehyde. Two running sutures of polygalactin 5-0 were applied, one on each side of the patch. The corpus spongiosum was closed with uninterrupted suture and the skin with interrupted suture of polygalactin 5-0. Six months later, the animals were examined and sacrificed under anesthesia. Retrograde urethrograms showed that the urethral healing was complete in six of the 30 animals, without stenosis, fistulas or dilations. Microscopic examination showed complete epithelization of these six urethras. The remaining 24 animals presented urethrocutaneous fistulas without stenosis, demonstrated by urethral catheterism using a 10-Fr plastic catheter. These data show that a successful urethral reconstruction of the penile urethra was possible in only 20% of the operated animals. Infection and leakage may be the cause of the urethrocutaneous fistulas present in 80% of cases. Further studies are necessary to determine whether such fistulas are avoidable. If they are, the bovine pericardium may well be an option in the treatment of urethral lesions in dogs.

Nocardia otitidiscaviarum (GAM-5) induces parkinsonian-like alterations in mouse

Parkinson's disease, a major neurodegenerative disorder in humans whose etiology is unknown, may be associated with some environmental factors. Nocardia otitidiscaviarum (GAM-5) isolated from a patient with an actinomycetoma produced signs similar to Parkinson's disease following iv injection into NMRI mice. NMRI mice were infected intravenously with a non-lethal dose of 5 x 10(6) colony forming units of N. otitidiscaviarum (GAM-5). Fourteen days after bacterial infection, most of the 60 mice injected exhibited parkinsonian features characterized by vertical head tremor, akinesia/bradykinesia, flexed posture and postural instability. There was a peak of nocardial growth in the brain during the first 24 h followed by a decrease, so that by 14 days nocardiae could no longer be cultured. At 24 h after infection, Gram staining showed nocardiae in neurons in the substantia nigra and occasionally in the brain parenchyma in the frontal and parietal cortex. At 21 days post-infection, tyrosine hydroxylase immunolabeling showed a 58% reduction of tyrosine hydroxylase in the substantia nigra, and a 35% reduction of tyrosine hydroxylase in the ventral tegmental region. Dopamine levels were reduced from 110 ± 32.5 to 58 ± 16.5 ng/mg protein (47.2% reduction) in brain from infected mice exhibiting impaired movements, whereas serotonin levels were unchanged (191 ± 44 protein in control and 175 ± 39 ng/mg protein in injected mice). At later times, intraneuronal inclusion bodies were observed in the substantia nigra. Our observations emphasize the need for further studies of the potential association between Parkinson's disease or parkinsonism-like disease and exposure to various nocardial species.

Monoamines in the pedal plexus of the land snail Megalobulimus oblongus (Gastropoda, Pulmonata)

In molluscs, the number of peripheral neurons far exceeds those found in the central nervous system. Although previous studies on the morphology of the peripheral nervous system exist, details of its organization remain unknown. Moreover, the foot of the terrestrial species has been studied less than that of the aquatic species. As this knowledge is essential for our experimental model, the pulmonate gastropod Megalobulimus oblongus, the aim of the present study was to investigate monoamines in the pedal plexus of this snail using two procedures: glyoxylic acid histofluorescence to identify monoaminergic structures, and the unlabeled antibody peroxidase anti-peroxidase method using antiserum to detect the serotonergic component of the plexus. Adult land snails weighing 48-80 g, obtained from the counties of Barra do Ribeiro and Charqueadas (RS, Brazil), were utilized. Monoaminergic fibers were detected throughout the pedal musculature. Blue fluorescence (catecholamines, probably dopamine) was observed in nerve branches, pedal and subepithelial plexuses, and in the pedal muscle cells. Yellow fluorescence (serotonin) was only observed in thick nerves and in muscle cells. However, when immunohistochemical methods were used, serotonergic fibers were detected in the pedal nerve branches, the pedal and subepithelial plexuses, the basal and lateral zones of the ventral integument epithelial cells, in the pedal ganglion neurons and beneath the ventral epithelium. These findings suggest catecholaminergic and serotonergic involvement in locomotion and modulation of both the pedal ganglion interneurons and sensory information. Knowledge of monoaminergic distribution in this snail´s foot is important for understanding the pharmacological control of reflexive responses and locomotive behavior.

Striatal and extrastriatal dopamine D2/3 receptors studied with [11C]raclopride and high-resolution PET

The human striatum is a heterogeneous structure representing a major part of the dopamine (DA) system’s basal ganglia input and output. Positron emission tomography (PET) is a powerful tool for imaging DA neurotransmission. However, PET measurements suffer from bias caused by the low spatial resolution, especially when imaging small, D2/3 -rich structures such as the ventral striatum (VST). The brain dedicated high-resolution PET scanner, ECAT HRRT (Siemens Medical Solutions, Knoxville, TN, USA) has superior resolution capabilities than its predecessors. In the quantification of striatal D2/3 binding, the in vivo highly selective D2/3 antagonist [11C] raclopride is recognized as a well-validated tracer. The aim of this thesis was to use a traditional test-retest setting to evaluate the feasibility of utilizing the HRRT scanner for exploring not only small brain regions such as the VST but also low density D2/3 areas such as cortex. It was demonstrated that the measurement of striatal D2/3 binding was very reliable, even when studying small brain structures or prolonging the scanning interval. Furthermore, the cortical test-retest parameters displayed good to moderate reproducibility. For the first time in vivo, it was revealed that there are significant divergent rostrocaudal gradients of [11C]raclopride binding in striatal subregions. These results indicate that high-resolution [11C]raclopride PET is very reliable and its improved sensitivity means that it should be possible to detect the often very subtle changes occurring in DA transmission. Another major advantage is the possibility to measure simultaneously striatal and cortical areas. The divergent gradients of D2/3 binding may have functional significance and the average distribution binding could serve as the basis for a future database. Key words: dopamine, PET, HRRT, [11C]raclopride, striatum, VST, gradients, test-retest.

Avaliação do relacionamento entre danos mecânicos e vigor, em sementes de milho, por meio da análise de imagens

A possibilidade de utilização da técnica de análise de imagens, para a determinação de danos mecânicos em sementes, é promissora; é um método de precisão, onde as sementes podem ser examinadas individualmente em imagens ampliadas e capazes de indicar, com detalhes, a área danificada, a localização e a extensão do dano. Por se tratar de método não destrutivo, as sementes em análise podem ser submetidas a testes fisiológicos e, desta forma, permitir o estabelecimento de relações entre os danos mecânicos e os prejuízos causados à qualidade. Assim, o presente trabalho teve o objetivo de estudar os efeitos dos danos mecânicos sobre o vigor de sementes de milho, utilizando a referida técnica. Cinqüenta sementes de cada um dos cultivares estudados (CO 32, AG 6690 e Attack) foram selecionadas visualmente, de tal maneira que a amostra fosse constituída por sementes íntegras e com danos mecânicos de intensidades variáveis. Em seguida, as sementes foram radiografadas, fotografadas (faces ventral e dorsal) e destinadas ao teste de frio (método do rolo de papel com terra). As plântulas normais, anormais e sementes mortas provenientes do teste de frio foram, também, fotografadas. As imagens obtidas foram disponibilizadas em computador de tal maneira que, examinadas simultaneamente na tela do monitor, pudessem ser buscadas relações de causa e efeito nas imagens anteriores e posteriores ao teste de frio. Os resultados obtidos indicam que o método estudado permite relacionar o dano mecânico com eventuais prejuízos ocasionados ao vigor das sementes de milho.

Análise de imagens na avaliação da qualidade de sementes de milho localizadas em diferentes posições na espiga

A análise de imagens, para avaliar a qualidade de sementes, é um método de precisão que possibilita examinar, com detalhes, a região danificada ou alterada, sua localização e extensão. Por ser um método não destrutivo, as sementes em análise podem ser submetidas a testes fisiológicos e, desta forma, é possível estabelecer as relações de causa e efeito. Assim, a pesquisa teve o objetivo de estudar os efeitos de diferentes posições da semente de milho na espiga, sobre a qualidade, utilizando-se a técnica de análise de imagens. Para tanto, as espigas do cultivar DKB 212 foram debulhadas de maneira que pudessem ser isoladas as sementes das posições proximal, intermediária e distal. As sementes foram caracterizadas fisicamente e avaliadas quanto ao vigor e sanidade. Paralelamente, as sementes foram radiografadas, fotografadas (faces ventral e dorsal) e destinadas ao teste de primeira contagem de germinação; as plântulas normais, anormais e sementes mortas foram, também, fotografadas. Todas as imagens obtidas foram disponibilizadas em computador para serem examinadas simultaneamente e, assim, fazer um diagnóstico completo para cada semente. As sementes das posições proximal e intermediária apresentam qualidade semelhante e superiores às da posição distal; sementes esféricas com eixos embrionários apresentando torções, porém não alterados, não tem sua qualidade diminuída, no entanto as alterações nos eixos embrionários identificados por manchas escuras e sem definição, presentes em maior quantidade na posição distal da espiga, são responsáveis pela redução da qualidade das sementes.

Hot cognition and activation of the medial prefrontal cortex: Self-regulating in contexts involving motivational pressures

The medial prefrontal cortex (mPFC) is involved in performance-monitoring and has been implicated in the generation of several electrocortical responses associated with self-regulation. The error-related negativity (ERN), the inhibitory Nogo N2 (N2), and the feedback-related negativity (FRN) are event-related potential (ERP) components which reflect mPFC activity associated with feedback to behavioural (ERN, N2) and environmental (FRN) consequences. Our main goal was to determine whether or not rnPFC activation varies as a function of motivational context (e.g., those involving performance-related incentives) or the use of internally versus externally generated feedback signals (i.e., errors). Additionally, we assessed medial prefrontal activity in relation to individual differences in personality and temperament. Participants completed a combination of tasks in which performance-related incentives were associated with task performance and feedback generated from internal versus external responses. MPFC activity was indexed using both ERP scalp voltage peaks and intracerebral current source density (CSD) of dorsal and ventral regions. Additionally, participants completed several questionnaires assessing personality and temperament styles. Given previous studies have shown that enhanced mPFC activity to loss (or negative) feedback, we expected that activity in the mPFC would generally be greater during the Loss condition relative to the Win condition for both the ERN and N2. Also, due to the evidence that the (vmPFC) is engaged in arousing contexts, we hypothesized that activity in the ventromedial prefrontal cortex (vmPFC) would be greater than activity in the dorsomedial prefrontal cortex (dmPFC), especially in the Loss condition of the GoNogo task (ERN). Similarly, loss feedback in the BART (FRN) was expected to engage the vmPFC more than the dmPFC. Finally, we predicted that persons rating themselves as more willing to engage in approach-related behaviours or to exhibit rigid cognitive styles would show reduced activity of the mPFC. Overall, our results emphasize the role of affective evaluations of behavioural and environmental consequences when self-regulating. Although there were no effects of context on brain activity, our data indicate that, during the time of the ERN and N2 on the MW Go-Nogo task and the FRN on the BART, the vrnPFC was more active compared to the dmPFC. Moreover, regional recruitment in the mPFC was similar across internally (ERN) and externally (FRN) generated errors signals associated with loss feedback, as reflected by relatively greater activity in the vmPFC than the dmPFC. Our data also suggest that greater activity in the mPFC is associated with better inhibitory control, as reflected by both scalp and CSD measures. Additionally, deactivation of the subgenual anterior cingulate cortex (sgACC) and lower levels of self-reported positive affect were both related to increased voluntary risk-taking on the BART. Finally, persons reporting higher levels of approach-related behaviour or cognitive rigidity showed reduced activity of the mPFC. These results are in line with previous research emphasizing that affect/motivation is central to the processes reflected by mediofrontal negativities (MFNs), that the vmPFC is involved in regulating demands on motivational/affective systems, and that the underlying mechanisms driving these functions vary across both individuals and contexts.

The Cenozoic Gonyaulacacean Dinoflagellate Genera Operculodinium Wall, 1967 and Protoceratium Bergh, 1881 and Their Phylogenetic Relationships

The palynology of Ocean Drilling Program Site 1007, leeward of the present Bahamas Bank, provides insights into upper Oligocene–lower Pleistocene dinoflagellate cyst associations in the tropical Americas. These associations are reviewed along with the sedimentary paleoenvironment to provide context for a morphological study of the cystdefined dinoflagellate Operculodinium bahamense and its comparison with the thecadefined dinoflagellate Protoceratium reticulatum which produces a cyst assignable to the cyst-defined genus Operculodinium. Detailed reconstructions of the tabulation in both species reveal strong similarities, having a sexiform hyposomal tabulation and L-type or modified L-type ventral organization. Protoceratium reticulatum has dextral torsion of the hypotheca, requiring assignation of the genus to the subfamily Cribroperidinioideae, whereas Operculodinium bahamense has neutral torsion requiring assignation to the subfamily Leptodinioideae. Results either imply polyphyletic origins for the genus Operculodinium or that combinations of ventral organization and torsion cannot always be applied rigidly to subdivide the family Gonyaulacaceae.

Investigation of the role of the nucleus accumbens in amphetamine-induced 50 kHz ultrasonic vocalizations in the rat

There is extensive evidence that the mesolimbic dopamine system underlies the production of 50 kHz ultrasonic vocalizations in rats. In particular, the shell of the nucleus accumbens is associated with generation of frequency modulated 50 kHz calls (a specific type of 50 kHz call which can be subdivided into various subtypes). There is also evidence that amphetamine administered systemically preferentially increases the proportion of trill and step calls compared to other frequency modulated 50 kHz subtypes. The purpose of this study was to investigate the effect of drug administration route and the role of the nucleus accumbens shell in amphetamine-induced 50 kHz call profile in the rat. Three experiments investigated this by using subcutaneous and intra-accumbens microinjections of amphetamine, as well as procaine (a local anesthetic) blockade of the nucleus accumbens. Ultrasonic vocalizations were recorded digitally from 24 rats and were analysed for sonographic structure based on general call parameters. The results of the three experiments were partially supportive of the hypotheses. Systemic amphetamine was found to induce greater bandwidth in 50 kHz calling compared to spontaneous calls in a vehicle condition. Systemic amphetamine was also found to preferentially increase the proportion of trill and step subtypes compared to vehicle. Moreover, there was no difference in the proportions of 50 kHz subtypes resulting from intracerebral or systemic application of amphetamine. There was, however, a significant difference for bandwidth, with systemic amphetamine inducing greater bandwidth over intraaccumbens application. Procaine blockade of the nucleus accumbens shell paired with subcutaneous amphetamine produced no difference in bandwidth of calls compared with those after a vehicle pre-treatment similarly paired. There was no reduction in the proportions of trill and step 50 kHz subtypes as well, with the procaine condition showing significantly greater proportion of step calls. The results of the study support a role for the iii nucleus accumbens shell in the amphetamine-induced changes on 50 kHz call profile. They also indicate there are more regions and pathways involved in generating 50 kHz calls than the projections from the ventral tegmental area to the nucleus accumbens. The implications of this work are that frequency modulated 50 kHz subtypes may be generated by distinct neurophysiological mechanisms and may represent a profitable avenue for investigating different circuits of 50 kHz call categories in the rat.

Src family kinases are required for limb trajectory selection by spinal motor axons

Signal relay by guidance receptors at the axonal growth cone is a process essential for the assembly of a functional nervous system. We investigated the in vivo function of Src family kinases (SFKs) as growth cone guidance signaling intermediates in the context of spinal lateral motor column (LMC) motor axon projection toward the ventral or dorsal limb mesenchyme. Using in situ mRNA detection we determined that Src and Fyn are expressed in LMC motor neurons of chick and mouse embryos at the time of limb trajectory selection. Inhibition of SFK activity by C-terminal Src kinase (Csk) overexpression in chickLMCaxons using in ovo electroporation resulted inLMC axons selecting the inappropriate dorsoventral trajectory within the limb mesenchyme, with medial LMC axon projecting into the dorsal and ventral limb nerve with apparently random incidence. We also detected LMC axon trajectory choice errors in Src mutant mice demonstrating a nonredundant role for Src in motor axon guidance in agreement with gain and loss of Src function in chickLMCneurons which led to the redirection ofLMCaxons. Finally, Csk-mediated SFK inhibition attenuated the retargeting ofLMCaxons caused by EphA or EphB over-expression, implying the participation of SFKs in Eph-mediated LMC motor axon guidance. In summary, our findings demonstrate that SFKs are essential for motor axon guidance and suggest that they play an important role in relaying ephrin:Eph signals that mediate the selection of motor axon trajectory in the limb.

Étude du rôle de la dopamine et de la sérotonine dans l’effet atténuateur des antipsychotiques et de l’OSU-6162 sur la récompense induite par la stimulation du faisceau médian prosencéphalique chez le rongeur

La voie mésocorticolimbique est constitutée d’un ensemble d’éléments nerveux issus de l’aire tegmentaire ventrale mésencéphalique et projettant vers des régions corticales et sous-corticales. Les neurones à dopamine (DA) qui en font partie modulent plusieurs fonctions cognitives dont l’attention, l’apprentissage et la récompense. L’activité nerveuse des cellules à DA augmente lorsque l’organisme anticipe et reçoit une récompense, ainsi qu’au cours de la phase d’apprentissage des comportements d’appétence. Or, si l’activité dopaminergique de la voie mésocorticolimbique est désordonnée, voire aberrante, des stimuli neutres deviennent saillants et prennent une signification erronée. Cette anomalie fonctionnelle du système dopaminergique pourrait être à l’origine des symptômes psychotiques observés dans la schizophrénie. Cette hypothèse est renforcée par le fait que les médicaments antipsychotiques efficaces ont tous une activité antagoniste aux récepteurs à DA de type 2 (D2); les antipsychotiques dits classiques (i.e. halopéridole) possèdent une forte affinité pour les récepteurs D2 tandis que les antipsychotiques dits atypiques (i.e. clozapine) présentent une plus forte affinité pour les récepteurs à sérotonine de type 2a (5-HT2a) que pour les récepteurs D2. Les antipsychotiques atypiques semblent plus efficaces contre les symptômes négatifs (i.e. anhédonie) de la schizophrénie et induisent moins d’effets moteurs extrapyramidaux et de dysphorie que les antipsychotiques classiques. Il a été proposé que l’efficacité des antipsychotiques atypiques soit expliqué par leur double action antagoniste aux récepteurs 5-HT2a et D2. Afin de mieux comprendre les mécanismes de ces médicaments, nous avons étudié leurs effets sur la récompense en utilisant le modèle d’autostimulation intracérébrale (ASI) chez le rongeur. Le but de la première étude était d’évaluer l’effet d’un antagoniste sélectif des récepteurs 5-HT2a, le M100907, sur la récompense et sur l’atténuation de la récompense induite par l’halopéridole. L’hypothèse était que l’atténuation de la récompense induite par l’ajout du M100907 à l’halopéridole serait similaire à celle induite par la clozapine. Dans une seconde étude, l’effet sur la récompense d’un agoniste partiel aux récepteurs D2, l’OSU-6162, a été caractérisé sous deux conditions : i) en condition de base et ii) lorsque la neurotransmission dopaminergique est altérée par l’administration systémique de quinpirole, un agoniste des récepteurs D2/D3. Les hypothèses étaient que l’OSU-6162 i) atténuerait la récompense induite par la stimulation et ii) empêcherait l’atténuation et la facilitation de la récompense induites par le quinpirole. Les données obtenues montrent que le M100907 n’altère pas la récompense par lui-même mais réduit l’atténuation de la récompense induite par l’halopéridole. La co-administration du M100907 et de l’halopéridole induit une atténuation de la récompense d’amplitude similaire à celle induite par la clozapine, ce qui suggère que l’activité antagoniste aux récepteurs 5-HT2a de la clozapine contribue à son efficacité. Les données de la seconde étude montrent que l’OSU-6162 atténue la récompense, de manière dose-dépendante, ainsi que la facilitation, mais pas l’atténuation de la récompense induite par le quinpirole. Cette dernière observation suggère que l’OSU-6162 agit comme un antagoniste fonctionnel aux récepteurs D2 post-synaptiques. Un ensemble de données suggèrent que le comportement d’ASI constitue un modèle valide permettant d’évaluer l’efficacité antipsychotique potentielle de nouvelles molécules. Le comportement d’ASI est atténué par les antipsychotiques cliniquement efficaces mais est peu ou pas modifié par des molécules dépourvues d’activité antipsychotique. Les données obtenues dans cette thèse permettent de supposer que l’OSU-6162 possède une activité antipsychotique de nature atypique, et cela sans altérer la neurotransmission sérotoninergique.

Réorganisation fonctionnelle et structurale des cortex auditifs, visuels et associatifs chez les sourds profonds congénitaux ou prélinguaux

En raison de l’utilisation d’un mode de communication totalement différent de celui des entendants, le langage des signes, et de l’absence quasi-totale d’afférences en provenance du système auditif, il y a de fortes chances que d’importantes modifications fonctionnelles et structurales s’effectuent dans le cerveau des individus sourds profonds. Les études antérieures suggèrent que cette réorganisation risque d’avoir des répercussions plus importantes sur les structures corticales situées le long de la voie visuelle dorsale qu’à l’intérieur de celles situées à l’intérieur de la voie ventrale. L’hypothèse proposée par Ungerleider et Mishkin (1982) quant à la présence de deux voies visuelles dans les régions occipitales, même si elle demeure largement acceptée dans la communauté scientifique, s’en trouve aussi relativement contestée. Une voie se projetant du cortex strié vers les régions pariétales postérieures, est impliquée dans la vision spatiale, et l’autre se projetant vers les régions du cortex temporal inférieur, est responsable de la reconnaissance de la forme. Goodale et Milner (1992) ont par la suite proposé que la voie dorsale, en plus de son implication dans le traitement de l’information visuo-spatiale, joue un rôle dans les ajustements sensori-moteurs nécessaires afin de guider les actions. Dans ce contexte, il est tout à fait plausible de considérer qu’un groupe de personne utilisant un langage sensori-moteur comme le langage des signes dans la vie de tous les jours, s’expose à une réorganisation cérébrale ciblant effectivement la voie dorsale. L’objectif de la première étude est d’explorer ces deux voies visuelles et plus particulièrement, la voie dorsale, chez des individus entendants par l’utilisation de deux stimuli de mouvement dont les caractéristiques physiques sont très similaires, mais qui évoquent un traitement relativement différent dans les régions corticales visuelles. Pour ce faire, un stimulus de forme définie par le mouvement et un stimulus de mouvement global ont été utilisés. Nos résultats indiquent que les voies dorsale et ventrale procèdent au traitement d’une forme définie par le mouvement, tandis que seule la voie dorsale est activée lors d’une tâche de mouvement global dont les caractéristiques psychophysiques sont relativement semblables. Nous avons utilisé, subséquemment, ces mêmes stimulations activant les voies dorsales et ventrales afin de vérifier quels pourraient être les différences fonctionnelles dans les régions visuelles et auditives chez des individus sourds profonds. Plusieurs études présentent la réorganisation corticale dans les régions visuelles et auditives en réponse à l’absence d’une modalité sensorielle. Cependant, l’implication spécifique des voies visuelles dorsale et ventrale demeure peu étudiée à ce jour, malgré plusieurs résultats proposant une implication plus importante de la voie dorsale dans la réorganisation visuelle chez les sourds. Suite à l’utilisation de l’imagerie cérébrale fonctionnelle pour investiguer ces questions, nos résultats ont été à l’encontre de cette hypothèse suggérant une réorganisation ciblant particulièrement la voie dorsale. Nos résultats indiquent plutôt une réorganisation non-spécifique au type de stimulation utilisé. En effet, le gyrus temporal supérieur est activé chez les sourds suite à la présentation de toutes nos stimulations visuelles, peu importe leur degré de complexité. Le groupe de participants sourds montre aussi une activation du cortex associatif postérieur, possiblement recruté pour traiter l’information visuelle en raison de l’absence de compétition en provenance des régions temporales auditives. Ces résultats ajoutent aux données déjà recueillies sur les modifications fonctionnelles qui peuvent survenir dans tout le cerveau des personnes sourdes, cependant les corrélats anatomiques de la surdité demeurent méconnus chez cette population. Une troisième étude se propose donc d’examiner les modifications structurales pouvant survenir dans le cerveau des personnes sourdes profondes congénitales ou prélinguales. Nos résultats montrent que plusieurs régions cérébrales semblent être différentes entre le groupe de participants sourds et celui des entendants. Nos analyses ont montré des augmentations de volume, allant jusqu’à 20%, dans les lobes frontaux, incluant l’aire de Broca et d’autres régions adjacentes impliqués dans le contrôle moteur et la production du langage. Les lobes temporaux semblent aussi présenter des différences morphométriques même si ces dernières ne sont pas significatives. Enfin, des différences de volume sont également recensées dans les parties du corps calleux contenant les axones permettant la communication entre les régions temporales et occipitales des deux hémisphères.

Contribution of tachykinin and kinin receptors in central autonomic control of blood pressure and behavioural activity in hypertensive rats

This work aims at studing the role of tachykinin NK-3 receptor (R) and kinin B1R in central autonomic regulation of blood pressure (BP) and to determine whether the B1R is overexpressed and functional in rat models of hypertension by measuring the effect of a B1R agonist on behavioural activity. Assumptions: (1) NK-3R located in the ventral tegmental area (VTA) modulates the mesolimbic dopaminergic system and has a tonic activity in hypertension; (2) B1R is overexpressed in the brain of hypertensive rats and has a tonic activity, which contributes to hypertension via a dopamine mechanism; (3) the inhibition of NK-3R and B1R with selective antagonists, reduces central dopaminergic hyperactivity and reverses hypertension. A model of genetic hypertension and a model of experimental hypertension were used: spontaneously hypertensive rats (SHR, 16 weeks) and Wistar-Kyoto (WKY) rats infused for 14 days with angiotensin II (Ang II) (200 ng / kg / min, subcutaneous (s.c.) with Alzet mini pump). The age-matched untreated WKY rats served as common controls. In the first study (article # 1), the cardiovascular response in SHR was evaluated following intracebroventricular (i.c.v.) and/or intra-VTA injection of an agonist (senktide) and antagonists (SB222200 and R-820) of NK-3R. These responses have also been characterized using selective dopamine antagonists DA-D1R (SCH23390), DA-D2R (raclopride) or non-selective dopamine DA-D2R (haloperidol). Also the VTA has been destroyed by ibotenic acid. The pressor response induced by senktide and the anti-hypertensive response induced by SB222200 or R-820 were more pronounced by intra-VTA. These responses were prevented by pre-treatment with raclopride and haloperidol. The lesion of the VTA has prevented the pressor response relayed by senktide (i.c.v.) and the anti-hypertensive effect of R-820 (i.c.v.). In addition, SB222200 (intra-VTA) prevented the pressor response of senktide (i.c.v.) and conversely, senktide (i.c.v.) prevented the antihypertensive effect of SB222200 (intra-VTA). The second study (article # 2) showed that the B1R antagonist (SSR240612) administered by gavage or i.c.v. reverses hypertension in both models. This anti-hypertensive effect was prevented by raclopride and haloperidol. In contrast, the two B1R antagonists (R-715 and R-954) injected s.c., which do not cross the blood-brain barrier reduced weakly blood pressure in hypertensive rats. In the third study (article # 3), the i.c.v. injection of a selective kinin B1R agonist Sar[DPhe8][des-Arg9]BK caused behavioural responses in SHR and Ang II-treated rats and had no effect in control WKY rats . The responses elicited by B1R agonist were blocked by an antagonist of NK-1 (RP67580), an antagonist of NMDA glutamate receptor (DL-AP5), an inhibitor of nitric oxide synthase (NOS) (L -NNA) as well as raclopride and SCH23390.The responses were modestly affected by the inhibitor of inducible NOS (iNOS). The B1R mRNA (measured by RT-PCR) was significantly increased in the hypothalamus, the VTA and the nucleus accumbens of hypertensive animals (SHR and treated with Ang II) compared with control rats. These neuropharmacological studies suggest that: (1) the NK-3R from the VTA is involved in the maintenance of hypertension in SHR by increasing DA transmission in the midbrain; (2) the B1R in SHR and Ang II-treated rats contributes to hypertension via a central mechanism involving DA-D2R; (3) the central B1R increases locomotor activity and nocifensive behaviours via the release of substance P (NK-1), DA and nitric oxide in both rat models of hypertension. Thus, the brain tachykinin NK-3R and kinin B1R represent potential therapeutic targets for the treatment of hypertension. The modulation of the mesolimbic/mesocortical dopaminergic pathway by these receptors suggests their involvement in other physiological functions (pleasure, motor activity, coordination of the response to stress) and pathophysiology (anxiety, depression).

Régulation transcriptionelle du développement de l'hypothalamus chez l'amphibien

Le noyau paraventriculaire (PVN) de l'hypothalamus régule une série de phénomènes physiologiques incluant l'équilibre énergétique et la pression artérielle. Nous avons identifié une cascade de facteurs de transcription qui contrôle le développement du PVN. SIM1 et OTP agissent en parallèle pour contrôler la différenciation d'au moins cinq types de neurones identifiables par la production d'OT, AVP, CRH, SS et TRH. Ces Facteurs de transcriptions contrôlent le développement des lignées CRH, AVP et OT en maintenant l'expression de Brn2 qui à son tour est nécessaire pour la différenciation terminale de ces neurones. L'analyse du transcriptome du PVN nous a permis d'identifier plusieurs gènes qui ont le potentiel de contrôler le développement du PVN. Nous voulons développer un paradigme de perte de fonction qui permettrait l'étude de ces gènes candidats sur une grande échelle. Le but de ce projet est de caractériser le PVN en développement de l'amphibien en vue de l'utilisation de ce modèle pour des études fonctionnelles. Nous avons cloné des fragments de cDNA de Sim1, OTP, Brn2, Sim2, CRH, Ot, AVP et TRH à partir de l'ARN total de Xenopus Laevis. Nous avons adapté notre technique d'hybridation in situ pour caractériser l'expression de ces gènes chez l'amphibien aux stades 33-39, 44, 51, 54, 60, et chez l'adulte. Résultats. Les Facteurs de transcription Sim1, OTP, et Brn2 commencent à être exprimés dans le PVN prospectif au stade 33. L'expression des marqueurs de différenciation terminale devient détectable entre les stades 37 et 39. De façon intéressante, le PVN occupe initialement un domaine de forme globulaire puis à partir du stade 44 s'allonge le long de l’axe dorso-ventral. Cet allongement se traduit par une organisation en colonnes des cellules du PVN que nous n'avons pas observée chez les rongeurs. Le développement du PVN est conservé chez l'amphibien dans la mesure où la relation entre l'expression des facteurs de transcription et des marqueurs de différenciation terminale est conservée. Il existe par ailleurs des différences entre la topographie des PVN des mammifères et de l'amphibien. L'organisation en colonnes de cellules pourrait correspondre à des mouvements de migration tangentielle. Nous sommes maintenant en mesure de tester la fonction des facteurs de transcription dans le PVN par l'approche d'invalidation par morpholinos.

Expression des cotransporteurs cation-chlorure KCC2 et NKCC1 au cours du développement de la moelle épinière de l’opossum Monodelphis domestica

L’inhibition est nécessaire à la génération d’outputs coordonnés entre muscles antagonistes lors de la locomotion. Une baisse de la concentration neuronale en ions chlorure au cours du développement des mammifères conduit à l’émergence de l’inhibition. Cette baisse repose sur l’équilibre entre deux cotransporteurs cation-chlorure, KCC2 et NKCC1. KCC2 expulse Cl- de la cellule alors que NKCC1 pompe Cl- dans la cellule. L’opossum Monodelphis domestica naît dans un état très immature. Le seul comportement locomoteur qu’il présente à la naissance consiste en des mouvements rythmiques et alternés des membres antérieurs pour grimper le long du ventre de la mère vers une tétine. Les membres postérieurs sont des bourgeons immobiles dont le développement est en grande partie postnatal. Pour cette raison, cette espèce constitue un modèle idéal pour l’étude du développement locomoteur. Afin d’étudier les mécanismes conduisant à l’émergence de l’inhibition durant le développement moteur, nous avons décrit l’expression développementale de KCC2 et NKCC1 chez l’opossum postnatal par immunohistochimie au niveau des renflements spinaux. Les motoneurones et afférences primaires ont été identifiés en utilisant un marquage rétrograde au TRDA. Le marquage pour KCC2 et NKCC1 est détecté dans la moelle épinière ventrale dans la matière grise et blanche présomptive dès la naissance, ce qui suggère que l’inhibition serait déjà mise en place avant la naissance, permettant subséquemment l’alternance des membres antérieurs observée chez les nouveau-nés. L’expression développementale de KCC2 et NKCC1 suit des gradients ventrodorsal et médiolatéral, tels qu’observés chez les rongeurs (rats et souris). Le patron mature d’expression de ces cotransporteurs est observé aux alentours de la 5ème semaine postnatale lorsque la locomotion de l’opossum est mature. Enfin, entre la naissance et P5, les dendrites exprimant KCC2 au niveau de la corne dorsale sont retrouvées en apposition aux afférences primaires ce qui suggère un rôle de KCC2 dans la formation des circuits sensori-moteurs.