L’objectif principal de cette thèse est de comprendre l’origine de la variabilité d’expression du comportement sexuel (CS). Nous avons trouvé que l’approche latérale est le CS le plus exprimé et que sa quantification peut servir comme mesure de la libido et donner une idée de la performance. Nous avons montré des effets remarquables de l’âge et de la lumière sur le CS du bélier. Nous avons également montré que les structures cérébrales impliquées dans la phase motivationnelle et consommatoire ne étaient pas les mêmes et que quel que soit le partenaire sexuel préféré, l’exposition à ce partenaire provoquait une activation des mêmes structures cérébrales ; le niveau de libido semble être déterminé par des sites spécifiques et bien précis comme l’aire préoptique, le noyau paraventriculaire et la partie médiane du Lit de la strie terminale. Le Cortex ent pourrait fonctionner comme un inhibiteur de la motivation hétérosexuelle. / The main objective of this thesis was to acquire a better understanding of the origin of the variability of the expression of sexual behavior. The nudging is most expressed sexual behavior and quantification of this behavior can be used to measure their libido and give a clue on their performance. We have discovered the remarkable effects of age et lighting of the ram's sexual behavior and that the cerebral structures involved in the motivation phase and consummatory phase are not the same and seems that, no matter which sexual partner is chosen, the fact of being exposed to this partner provokes the activation of these cerebral structures et what the expression of sexual motivation level seems to have specific and precise sites in the Preoptic Area, the Paraventricular Nucleus (PVN) and the Nucleus of the bed of the striae terminalis medial (BNSTmed). The Entorhinal cortex could function as an inhibitor of heterosexual motivation.

Genetically based Central Nervous System (CNS) disorders remain a largely unresolved issue in the world today. Our genome is the source of our greatest strengths and weaknesses. For this reason, intelligent modification of the genome's DNA is a profoundly beneficial goal in the maximization of the overall health of the human race. Potential benefit in this field is currently limited in both effectiveness and safety in regards to the delivery of therapeutic genes into the nucleus, which is protected by many evolution-based barriers. Evolution has also favored the development of highly specialized and infectiously effective viruses capable of overcoming such boundaries. By neutering the naturally occurring and pathologically benign Adeno-Associated Virus (AAV) we have transformed what was once a virus, into a "pure" vector, taking full advantage of evolution's diligent enhancement of these genetic hijackers without introducing unacceptable danger to patients. We utilized the logically engineered, castrated form of AAV serotype 9 (recombinant AAV9/rAAV9) to act as a vehicle for two reporter genes, Enhanced Green Fluorescent Protein (EGFP) and Firefly Luciferase (Fluc) with the goal of assessing and improving the efficiency of vector transduction in murine CNS. We found that rAAV9, when infused into the intrathecal space of mice is capable of extensive and intensive transduction of both neurons and astrocytes throughout the entire length of the SC as well as the hind regions of the brain (brainstem and cerebellum). We also found that efficiency of transduction was best in our highest dose groups, 1E+12 genome copies (GC) in Experiment 1 and 2E+12 GC in Experiment 2, both of which received rAAV9 particles via the two commercially available (100μL and the 200μL) ALZET® Osmotic Pump designs. Administering dosage higher than this directly into the intrathecal space was limited by the size of the pump reservoir and rAAV9 production titer. We are currently attempting to achieve a more complete CNS transduction by performing another experiment in which we place the pump cannula directly into the intracerebroventricular (ICV) space of the lateral ventricles. Our findings reveal that infusion of rAAV9 by intrathecal placed pump cannula is more effective than any other method tested in this study int the transduction of neurons and glial cells of adult&ndashmouse CNS. By elucidating a mode of delivery that maximizes the robustness of transduction efficiency, our results are a critical building block in designing a cure for the array of genetic-based diseases of the CNS, which are currently untreatable

La L-Théanine (L-5-N-éthylglutamine) est un acide aminé présent dans le thé vert et qui a une structure similaire à celle de l'acide glutamique, le neurotransmetteur majoritaire du système nerveux central, SNC. La L-théanine possède la capacité de traverser la barrière hémato-encéphalique en plus de ses nombreuses activités physiologiques et pharmacologiques, anxiolytique et relaxante. La théanine et ses dérivés sont donc considérés comme des outils indispensables pour la compréhension de la synapse glutamatergique plus précisément. Nous avons synthétisé la théanine énantiomériquement pure (L et D). Nous avons développé deux nouvelles méthodes de synthèse de la théanine optiquement pure. Ensuite, nous avons décrit la préparation des dérivés 5-N-alkylés de la théanine et les dipeptides -glutamique en tant qu'analogues 5-N-substitués de la théanine. Dans le but d'élargir l'éventail d'applications de la Théanine ayant un intérêt biologique, nous avons synthétisé des dipeptides contenant la Théanine. Nous présentons également un accès aux dérivés 4-arylés de la théanine, via une alkylation régio et stéréosélective de l'acide pyroglutamique jamais décrit dans la littérature. L'analyse rigoureuse des intermédiaires réactionnels et des produits finaux par les RX, la RMN 1H à 600 MHZ et l'HRMS a prouvé l'obtention d'un seul diastéréoisomère (2S, 4R)-4-aryle théanineLes résultats des tests biologiques, utilisant la technique de l'imagerie calcique, montrent que les deux énantiomères L et D de la théanine possède un effet agoniste vis-à-vis les récepteurs NMDA et que cet effet est beaucoup plus important dans le cas de l'énantiomère (D). Parmi les analogues 5-N-alkylés de la théanine ayant un effet agoniste spécifique des récepteurs NMDA, la 5-N-Propyl-Gln (L et D) montre une activité en tant qu'agoniste beaucoup plus importante que la théanine même (naturelle ou synthétique) et aucun effet n'a été observé sur les récepteurs AMPA et métabotropiques. La L-théanine et ses dérivés pourraient donc être des structures intéressantes pour développer de nouveaux outils pharmacologiques nécessaires à l'étude des récepteurs glutamatergiques (métabotropiques et/ ou ionotropiques). / L-Theanine (5-N-L-ethylglutamine) which is an amino acid found in green tea, it has a structure similar to that of glutamic acid. L-theanine has the ability to cross the blood-brain barrier in addition to its physiological and pharmacological activities.Given the importance of this molecule as essential for the investigation of physiological roles of CNS tools, we synthesized the enantiomerically pure theanine (L and D). A serie of 5-N-substituted theanine were also synthesized. In order to broaden the range of applications of theanine, we synthesized dipeptides containing Theanine for the purpose of obtaining products that have biological significance. The regio and stereoselectively synthesized analogs of L-theanine in the 4-position substituted with an aryl group has been developed to be tested at the level of glutamate receptors.The results of biological tests, using calcium imaging technique, show that theanine with its two enantiomers (D and L) has an agonistic effect vis-à-vis the NMDA receptors and that this effect was much greater with the enantiomer (D). Among the 5-N-alkylated analogs of theanine which they had only an agonistic effect on the NMDA receptor, 5-N-Propyl-Gln (L and D) has activity as an agonist much larger than the theanine and no effects were scored on AMPA receptors and metabotropic. L-theanine and its derivatives could be interesting structures to develop new pharmacological tools to study glutamate receptors (metabotropic and / or ionotropic).

Group B Sox genes play a critical developmental role in both vertebrates and insects. Within the model species Drosophila melanogaster, two SoxB genes, Dichaete and SoxNeuro, have been shown to act as ‘master regulators’ in the early development of the central nervous system. SoxB genes have only been characterised in a handful of arthropod species thus far, with most work to date focusing on drosophilids. The purpose of this investigation was twofold. First, I set out to resolve the phylogenetic origins of arthropod SoxB genes, as mutually exclusive models explaining their emergence are still contested. I have identified and annotated the SoxB of several invertebrate taxa. In total, my investigation includes 24 different metazoan taxa, and represents the largest investigation of arthropod SoxB phylogeny to date. In light of this research, I have proposed a new model of SoxB evolution which resolves the conflicting elements of the two primary competing models. Second, to study the evolution of SoxB in terms of functional conservation/divergence, I selected the emerging model organism Tribolium castaneum to draw a comparative analysis with Drosophila melanogaster. I first began by characterising the spatiotemporal expression patterns of SoxNeuro mRNA in early Tribolium embryos using whole mount in situ hybridisation, and examined published Dichaete expression patterns in the context of central nervous system development in T. castaneum. Using these data, I draw a comparison to the expression profiles of Dichaete and SoxNeuro orthologues in Drosophila melanogaster and other species. I have found that both Dichaete and SoxNeuro expression patterns in the developing central nervous system are remarkably well-conserved across species. I also attempted to characterise genome-wide binding for both Dichaete and SoxNeuro proteins in Tribolium in what would have represented the first genomic investigation of its kind in this emerging species. Using a tethered DNA adenine methyltransferase (Dam) enzyme for both SoxNeuro and Dichaete, I hoped to characterise the genomic loci with which each protein interacts within the beetle genome (a technique known as DamID). Unfortunately, these last set of experiments have proved unsuccessful, despite several attempts which have made use of different promoters, different DNA enrichment methodologies, and tackling unforeseen DNA contamination issues. Nevertheless, the troubleshooting experiments that I have carried out will pave the way for further genomic experiments in Tribolium, easing the establishment of genomic research in this emerging organism.

Orientador: Man Chin Chung / Coorientador: Carlos Cesar Crestani / Banca: Silvia Honda Takada / Banca: Ellen Cristini de Freitas / Banca: Priscila Longhin Bosquesi de Oliveira / Banca: Eduardo Rene Perez Gonzales / Resumo: O consumo excessivo de etanol está relacionado a diminuição da neurogênese hipocampal e a diversas patologias cardiovasculares, como a hipertensão. A taurina é um aminoácido não-essencial encontrado principalmente em estruturas cujos tecidos são mais excitáveis, como músculo esquelético, tecido cardíaco, vasos sanguíneos e sistema nervoso central. Níveis normais deste nutriente são essenciais para o correto funcionamento do organismo e sua depleção pode precipitar diversos quadros patológicos. A taurina tem apresentado resultados promissores tanto no sistema cardiovascular quanto no sistema nervoso central. Desta maneira, o objetivo deste trabalho foi estudar os efeitos da taurina na função cardiovascular bem como na neurogênese hipocampal de ratos submetidos ao modelo de consumo forçado de etanol. Dessa forma, ratos Wistar de aproximadamente 250 gramas receberam soluções crescentes de etanol (5% na primeira semana, 10% na segunda semana e 20% na terceira e quarta semana), sem a oferta de água; o grupo controle recebeu apenas água. A taurina foi administrada (i.p., 300 mg/kg) diariamente durante 28 dias e os animais do grupo controle receberam injeção apenas do veículo. O presente trabalho foi dividido em dois experimentos. No primeiro, a taurina foi administrada juntamente com a oferta de etanol. Já o segundo experimento, os animais foram expostos ao etanol e depois, receberam a taurina. Os resultados do Experimento 1 não mostraram alterações significativas na função cardiov... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Excessive ethanol consumption is related with decrease in hippocampal neurogenesis and various cardiovascular disorders such as hypertension. Taurine is a non-essential amino acid and is found primarily in structures which are more excitable tissues, such as skeletal muscle, heart tissue, blood vessels and central nervous system. Normal levels of this compound are essential for the correct functioning of the body and its depletion may precipitate various pathological conditions. Among the various compounds studied to reverse or protect against the effects of ethanol, taurine have shown promising results both in the cardiovascular system and the central nervous system. Thus, the objectives of this study was to investigate the effects of taurine on cardiovascular function and hippocampal neurogenesis in rats submitted to the model of forced consumption of ethanol. Wistar rats of 250g of body weight received ethanol solution (5% in the first week, 10% in the second week and 20% in the third and fourth week) without the offer of water. Taurine was administered (i.p., 300 mg / kg) daily for 28 days. This study was divided into two experiments. In the first one, to analyze the protective effects of taurine on the cardiovascular system and hippocampal neurogenesis against the effects of ethanol consumption, taurine was administered along with the ethanol consumption model. In the second experiment, to analyze the effects of taurine in reversing the effects of ethanol consumption, the animals were exposed to the ethanol consumption model and then received taurine. The results of Experiment 1 showed no significant changes in cardiovascular function... (Complete abstract click electronic access below) / Doutor

Lysosomal storage disorders (LSDs) comprise a broad disease group of inherited metabolic disorders, the majority of which are associated with CNS pathology, significant disability and reductions in life expectancy. LSDs are caused by mutations in genes encoding proteins critical for the correct functioning of lysosomal homeostasis. The disruption of lysosomal homeostasis results in the abnormal accumulation of lysosomal content, initiating intracellular pathological events, including calcium dysregulation and lysosomal membrane permeablisation (LMP) affecting cell function and inducing cellular death mechanisms. These pathological events are particularly damaging within the CNS, due to its limited capacity for regeneration. Despite intensive scientific research into these disorders, and an increased understanding of the pathological events underlying these diseases, effective treatments are still lacking for most LSDs. Several therapeutic approaches have been investigated in the last 30 years, including enzyme replacement therapy, bone marrow transplantation, substrate reduction therapy, chemical chaperones and gene therapy. However, the CNS pathology in many of the LSDs remains unaddressed due to the restricted ability of many therapeutic agents to cross the blood-brain barrier. The heat-shock response (HSR) is an emerging element involved in the pathogenesis of a variety of disorders. The HSR is a physiological response to a wide range of cellular stresses. It functions to protect the cell from the aggregation of misfolded proteins and LMP. Of the HSR, several key players are integral to mounting a heat shock response, these include the heat-shock factor 1 (HSF-1) and HSP70. In this thesis, we provide proof-of-principle for the use of recombinant HSP70, and the small molecule up-regulator of the HSR, arimoclomol in treatment of a range of LSDs. We show that HSP70 is able to access the CNS, and increase the degradative capacity of lysosomal hydrolases. This provides differential behavioural, biochemical and survival effects in LSD models of Niemann-Pick type C, Sandhoff and Fabry disease. Additional studies using the HSF-1 upregulator arimoclomol, show a complex dose-response between the different models, possibly reflecting essential differences in the calcium dysregulation between these disease states.

Caffeine consistently enhances endurance performance in temperate environmental conditions, while far less research has examined its ergogenic and physiological effects during prolonged exercise in the heat. Despite the performance benefit of an acute caffeine doses being less pronounced in regular caffeine users versus those not habituated to the drug, few studies have examined the influence of a prolonged period of controlled caffeine intake on endurance performance. The endogenous trace amine octopamine is purported to possess stimulant-like properties and influence fat metabolism, although no study has examined these effects in humans. The aim of this thesis was to further characterise the performance and physiological effects of caffeine during prolonged exercise, while elucidating a potential ergogenic role for octopamine. The first two studies investigated the ergogenic and thermoregulatory effects of low to moderate caffeine doses during prolonged cycle exercise in the heat. Chapter 4 demonstrated that 3 mg kg-1 caffeine, administered either as a single or split-dose (2 x 1.5 mg kg-1) before exercise, improved endurance performance without influencing thermoregulation during prolonged exercise at a fixed work-rate. Dividing the caffeine bolus appeared to confer an additional performance benefit, suggesting repeated low dose may potentiate the efficiency of the same total caffeine dose under these conditions. Chapter 5 demonstrated that a 6 mg kg-1 caffeine dose improved endurance cycle performance without differentially influencing thermoregulation than placebo. The level of habituation to caffeine influences the ergogenic effect of an acute dose, yet previous studies have employed sub-chronic supplementation protocols. Chapter 6 investigated the effect of a twenty-eight day supplementation period on endurance cycle performance. Habituation to caffeine attenuated the ergogenic effect of an acute caffeine dose, without any change in circulating caffeine, substrate oxidation or hormonal concentrations. In chapter 7 the performance and metabolic effects of octopamine was investigated. Octopamine supplementation did not influence performance, hormonal concentrations or substrate oxidation, likely due to low serum concentrations of the drug.

xiii, 104 p. : ill. (some col.) A print copy of this thesis is available through the UO Libraries. Search the library catalog for the location and call number. / Neurogenesis in Drosophila and mammals requires the precise integration of spatial and temporal cues. In Drosophila, embryonic neural progenitors, called neuroblasts, sequentially express the transcription factors Hunchback, Kruppel, Pdml/Pdm2 (Pdm) and Castor as they divide to generate a stereotyped sequence of neuronal and glial progeny. Hunchback is necessary and sufficient to specify the firstborn cell identity in many neuroblast lineages. Additionally, Hunchback is able to maintain an early-competence state in which early-born cells are generated. Furthermore, the Hunchback mammalian ortholog, Ikaros, possesses a similar ability to specify early- born cells in the vertebrate nervous system. However, the mechanisms underlying the function of Hunchback/Ikaros are unknown. Pdm and Castor are expressed later in many neuroblasts and can specify late-born neuronal cell identities in a model neuroblast lineage, NB7-1. Previous work studying their function in the NB7-1 lineage showed that Pdm and Castor act as repressors of Kruppel gene expression and inhibit the generation of the Kruppel-dependent cell identity. It is not known if the functions of Pdm and Castor are conserved across multiple neuroblast lineages during neurogenesis or whether these factors impart any restrictions on the ability of a factor like Hunchback to maintain early competence. To investigate the transcriptional mechanisms regulating early neuroblast competence in Drosophila, I have focused my dissertation research on two aims. The first is to examine the function of Pdm and Castor across multiple neuroblast lineages to characterize their potential roles as competence restricting factors; the second is to determine how Hunchback maintains early neuroblast competence and specifies early-born cell identities (e.g. as a transcriptional activator, repressor, or both). My work demonstrates that Pdm and Castor control the timing of Kruppel gene expression, and possibly the timing of other genes, in neuroblasts. Furthermore, I have shown that Hunchback acts as a transcriptional repressor of multiple target genes, including pdm and castor, to maintain early neuroblast competence. Because Hunchback must repress at least one additional unknown factor that can restrict neuroblast competence, I have piloted a screen to identify and characterize novel Hunchback target genes in the nervous system. This dissertation includes previously published and unpublished co-authored materials. / Committee in charge: Victoria Herman, Chairperson, Biology; Christopher Doe, Advisor, Biology; Judith Eisen, Advisor, Biology; Charles Kimmel, Member, Biology; Hui Zong, Member, Biology; Kenneth Prehoda, Outside Member, Chemistry

The present study aimed at investigating the neuropsychological sequelae of transient ischaemic attacks. Transient ischaemic attacks are defined as those neurological disorders in which there is complete resolution of neurological symptoms within twenty·four hours. Transient ischaemic attacks may or may not reveal evidence of brain infarcts on imaging studies. In the present study, the neuropsychological sequelae of transient ischaemic attacks in the carotid circulation were investigated since, within the perspective of cognitive neuropsychology, it was assumed that localized changes in cognitive functions could be demonstrated.Since several psychological, medical and neurological factors are known to influence scores·on neuropsychological tests, regression analyses were performed to determine which factors contributed significantly to the variance of scores on neuropsychological tests in the transient ischaemic attack and control groups. Two transient ischaemic attack groups, each comprising forty left and forty right hemisphere involvement patients, were then compared with each other and with a control group of forty general medical patients. Stenosis of the carotid artery formed a significant predictor of test scores in the combined transient ischaemic attack group. When the groups were·analyzed independently, in the left transient ischaemic attack group stenosis predicted performance on the same tests reaching significance for the combined group, and for the Wisconsin Card Sorting Test (Perseverative Score). In the right transient ischaemic attack group, stenosis significantly predicted performance on Digits Forward, Backward and Total, the PASAT (2.4 seconds) and Trails B. On the other hand, education formed a significant predictor of performance on Digits Forward, Digits Backward and Digits Total and the PASAT (all levels) in the control group. Multivariate comparisons revealed that the left and right transient ischaemic attack groups performed worse than the controls on tests of attention, concentration and conceptual flexibi1ity. The left transient ischaemic attack group performed worse than the right transient ischaemic attack group on all tests of attention and concentration, but there was a significantly better performance of the former group on the Rey Auditory Verbal Learning Test (Trial 1), Block Designs and Verbal Fluency. The findings on the PASAT that left transient ischaemic attack patients performed significantly worse than the right hemisphere group ·were considered to be relatively unreported previously in the literature on transient ischaemic attacks. The findings obtained are discussed from a neurocognitive perspective of neuropsychological functioning in transient ischaemic attacks. / Psychology / Ph. D. (Psychology)

Made available in DSpace on 2015-12-10T14:22:56Z (GMT). No. of bitstreams: 0 Previous issue date: 2015-07-31. Added 1 bitstream(s) on 2015-12-10T14:29:07Z : No. of bitstreams: 1 000853245.pdf: 5848716 bytes, checksum: 9fb857dfa29e26d6bcd61db677d6bdc7 (MD5) / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) / Os neurônios da raiz coclear (CRN) são os primeiros neurônios do sistema nervoso central a receber a informação auditiva e conectam-se com centros de integração sensoriomotora do tronco encefálico, como o núcleo reticular caudal da ponte. Os CRN estão envolvidos no circuito elementar do reflexo auditivo de sobressalto (RAS) juntamente com os neurônios do gânglio espiral, o núcleo reticular caudal da ponte e os motoneurônios da medula espinal. O RAS apresenta uma série de modulações como a habituação, a sensibilização, a potenciação por um estímulo adverso e a inibição por estímulo prévio (PPI). As modulações do RAS ocorrem mediante a influência de diversos núcleos sobre os componentes do circuito elementar deste reflexo e possuem valor diagnóstico na clinica médica de enfermidades neurodegenerativas e psiquiátricas como Parkinson e esquizofrenia. O núcleo da raiz coclear é o menos estudado do circuito neural do RAS. Pouco se conhece sobre as origens das aferências aos CRN e a identidade neuroquímica das mesmas. Recentemente foi descrita uma nova fonte de aferências aos neurônios da raiz coclear, de natureza colinérgica e proveniente do núcleo ventral do corpo trapezóide (VNTB). Os neurônios deste núcleo recebem informação auditiva diretamente do colículo inferior que faz parte da via auditiva e também participa do circuito neuronal responsável por mediar a PPI. Acredita-se que o colículo inferior possa mediar a PPI também ao nível do núcleo da raiz coclear via a conexão com o VNTB, o que constituiria uma via descentente curta, com participação na PPI com intervalos interestímulos curtos. Contudo, não existem estudos a respeito de aspectos funcionais desta conexão. Neste trabalho estudamos o papel da projeção colinérgica proveniente do VNTB através da lesão seletiva deste núcleo e avaliação comportamental do RAS e da PPI. Para isto realizamos 3 grupos experimentais: Controle... / The cochlear root neurons (CRN) are the first in the central nervous system to receive the acoustic information. They are in connection with sensory integration centers in the brainstem, particularly the pontine caudal nucleus. The CRN are related with the fundamental neural circuit of the acoustic startle reflex, together with the spiral ganglion neurons, the pontine reticular nucleus and the spinal cord motoneurons. The acoustic startle reflex shows various modulations such as habituation, sensitization, fear potentiation and prepulse inhibition. The modulations of the acoustic startle reflex are important in clinical diagnostics of psychiatric and neurodegenerative illnesses such as schizophrenia and Parkinson's disease. Startle modulations are promoted by influences from several nuclei on the fundamental neural circuit of the startle reflex. The cochlear root nucleus is the least studied nucleus of this circuit. The origins of the afferents to cochlear root neurons and its neurochemical identity remain unclear. Recently was described a new source of afferents to cochlear root neurons, cholinergic and coming from the ventral nucleus of the trapezoid body (VNTB). The neurons of this nucleus receive auditory information directly from the inferior colliculus which take part of the auditory pathway and also participates in the neural circuitry responsible for mediating the prepulse inhibition. Therefore, the inferior colliculus may mediate the prepulse inhibition at the cochlear root level via ventral nucleus of the trapezoid body connection, which would be a short and low latency pathway. However, there are no studies on functional aspects of this connection. We intend to study the role of cholinergic projections from the ventral nucleus of the trapezoid body by selective lesion of this nucleus and behavioral assessment of auditory startle reflex and prepulse inhibition. To achieve the purpose we proposed 3 experimental groups: Control ... / FAPESP: 2013/25737-6