Smith-Magenis Syndrome (SMS) [OMIM, #182290] is a congenital anomaly and mental retardation (MCA/MR) syndrome associated with deletion of chromosome17p11.2 [1]. The clinical phenotype has been well described and includes minor craniofacial anomalies, self-injurious behaviors as well as sleep disturbances, speech delays, and obesity [1,2,3]. The incidence of SMS is estimated to be ~ 1:15,000 - 25,000 births [2,6]. Among SMS patients, ~90% are comprised of 17p11.2 deletions, while ~10% have RAI1 mutations [8]. All 17p11.2 deletions associated with SMS include RAI1 deletion [10]. RAI1 is thought to function as a transcriptional factor although its cellular role is still unclear. First, in order to better understand the role of RAI1 as a transcriptional factor and its relation to SMS, we confirmed that RAI1 regulates BDNF within an intronic region. This sequence was further narrowed down by utilizing the luciferase reporter assay. This test confirmed what was previously found using ChIP-chip assay and microarray analysis of Rai1+/- mice hypothalami. Next, in order to evaluate the role of Bdnf, an ampakine drug was administered to the Rai1+/- mouse model. A mouse model is a powerful tool for studying a specific gene. Rai1+/- mice exhibit the SMS phenotypes of obesity, craniofacial abnormalities, reduced pain sensitivities, seizures and others. Many physical, neurological, and behavioral tests were performed on the mice to see if any of the phenotypes can be rescued. Interestingly, twice-daily injections of ampakine CX1837 restored the pain sensitivities in Rai1+/- mice. The hot plate data suggest that BDNF potentially has a role in regulating the SMS phenotype of decreased pain sensitivity. In order to evaluate other genes that are altered as a result of the CX1837 ampakine drug, the whole brain's global gene expression was evaluated via microarray analysis. Two potential pain-related genes were identified to be upregulated due to drug administration, which could account for the pain phenotypes observed. One of the genes upregulated in treated mice was Osm, which is interesting because Osm is responsible for pain sensitivity. Further analysis is needed to confirm that an ampakine drug can potentially be used to treat SMS patients.

Untersuchung zu single-nucleotid-polymorphismen (SNPs) der Serotoninrezeptoren 5-HT2A, 5-HT3A, des Neurotrophins BDNF und des Enzyms Tryptophanhydroxlase auf deren Korrelation mit Migräne mit und ohne Aura. / Correlation between single nucleotide polymorphisms in genes of serotonergic 5-HT2A and 5-HT3A-receptors, brain derived neurotropic factor and Tryptophanhydroxylase with migraine with and without aura phenomenon.

Schwermut, Einsamkeit, Desinteresse: 2-7% der Weltbevölkerung werden von diesen Gefühlen bestimmt, sie sind an einer Depression erkrankt. In Deutschland leiden aktuell bis zu 4 Mio. Menschen an einer Depression. Zwei Drittel diese Kollektivs befinden sich in hausärztlicher Behandlung, aber nur etwa die Hälfte von diesen Patienten wird korrekt diagnostiziert und behandelt. Die Gründe hierfür sind auf der einen Seite die schwierige Diagnosefindung, auf der anderen die bisher nicht vollständig geklärte Ätiologie und die komplexen Wirkmechanismen der medikamentösen antidepressiven Therapieoptionen. Die Entstehung einer depressiven Episode ist multifaktoriell bedingt, wobei insbesondere genetischen Faktoren in der Pathogenese depressiver Erkrankungen eine wichtige Rolle zugeordnet wird. Neurobiologische Untersuchungen der molekularen und biochemischen Hintergründe von depressiven Episoden befassen sich zurzeit insbesondere mit drei Hypothesen: Diese sind die Neurotransmitter-Dysbalance-Hypothese (Monoaminhypothese), die Stressachsen-Hypothese (Hypothalamus-Hypophysen-Achse) und die Neurotrophin-Hypothese. Die Stressachsen-Hypothese befasst sich in diesem Zusammenhang mit der Aktivität des Hypothalamus-Hypophysen-Systems, die bei depressiv erkrankten Patienten dysreguliert ist und mit einer erhöhten Kortisolsekretion einhergeht. Die Monoaminhypothese der Depression postuliert eine Dysfunktion serotonerger, noradrenerger und dopaminerger Systeme. Die Neurotrophinhypothese bezieht sich auf das Second Messenger-System des durch Antidepressiva aktivierten cyclischen Adenosin-Monophosphat (cAMP) Signalwegs. Cyclisches Adenosin-Monophosphat aktiviert die cAMP-abhängige Proteinkinase (PKA), die wiederum den Transkriptionsfaktor cAMP response element binding protein (CREB) phosphoryliert und ihn somit stimuliert. Aktiviertes CREB verstärkt die Transkription zahlreicher Zielgene, inklusive die des brain-derived neurotrophic factor (BDNF), welcher unter anderem als Regulator von Neurotransmittern dient und Überleben, Differenzierung und Plastizität von Neuronen beeinflusst. Astrozyten gehören zur Gruppe der Makrogliazellen im zentralen Nervensystem (ZNS) und sind die größte Population der Gliazellen. Sie sind für die Kaliumhomöostase und ebenso für die Regulation der synaptischen Transmission und der neurovaskulären Koppelung zuständig. Des Weiteren scheinen Astrozyten eine wichtige Rolle für die Bildung glialer Narben, die Induktion der Blut-Hirn-Schranke und auch für das neuronale Überleben zu spielen. Bei der Analyse der Wirkmechanismen medikamentöser antidepressiver Therapien ist in der letzten Zeit die Rolle der Astrozyten in den Vordergrund gerückt, um deren Beitrag zu antidepressiven Therapien zu untersuchen. Das Ziel der vorliegenden Arbeit war zu untersuchen, ob in Astrozyten der Serotonin-Transporter (5-HT-Transporter, 5-HTT), der Brain-derived neurotrophic factor (BDNF), der Dopamin-Transporter oder die Stickstoffmonoxyd-Synthase III (endotheliale Stickstoffmonoxid-Synthase, eNOS) gebildet werden und falls ja, ob sich deren Expression durch Applikation von Antidepressiva ändert. Die aus Rattenhirnen gewonnenen Astrozytenkulturen wurden hierfür entweder mit dem trizyklischen Antidepressivum Imipramin, dem selektiven Serotoninrückaufnahmeinhibitor Escitalopram oder zur Kontrolle mit Kochsalzlösung inkubiert. Nachdem die entsprechende mRNA aus den behandelten Astrozytenkulturen extrahiert wurde, ist sie in cDNA translatiert und mit Hilfe der quantitativen Real-Time PCR quantifiziert worden. Durch Vergleich der Expression des zu untersuchenden Gens mit der Expression der Housekeeping-Gene 18s-rRNA, Glycerinaldehyddehydrogenase (GAPDH) und Acidic ribosomal phosphoprotein (ARP) wurden Ungenauigkeiten bei der cDNA-Synthese ausgeglichen und die Daten normalisiert. Die rechnerische Auswertung der quantitativen Real-Time PCR erfolgte unter Verwendung der Ct-Werte unter Zuhilfenahme der geNORM Software. Die Ergebnisse zeigen eine signifikant erhöhte BDNF-Expression nach Imipramingabe. Hierbei zeigen bei den getrennten Untersuchungen der jeweiligen mRNA Chargen die mit 100µM Imipramin behandelten Astrozytenkulturen stärker signifikante Ergebnisse, als die mit 50µM Imipramin behandelten Astrozytenkulturen. Werden alle Proben, die mit der jeweiligen Imipraminkonzentration 4 Stunden inkubiert wurden, zusammen analysiert und mit den jeweiligen Kontrollen verglichen, zeigt sich sowohl bei der Behandlung mit 50µM Imipramin als auch mit 100µM Imipramin eine signifikante Steigerung der BDNF Expression. Escitalopram stimulierte die BDNF-Expression zwar ebenfalls nominell, jedoch war der Effekt nicht signifikant. Des Weiteren konnte eine deutliche Expression von 5-HTT-mRNA in Astrozytenkulturen nachgewiesen werden. Jedoch hatte keines der Antidepressiva einen signifikanten Effekt auf die 5-HTT-Expression. Es konnten weder in den behandelten, noch in den unbehandelten Astrozytenkulturen DAT oder NOS-III nachgewiesen werden. / Melancholia, loneliness, lack of interest: 2-7% of world population are ruled by these feelings. In Germany, 4 million inhabitants suffer from depression. Two third of this population are under medical treatment, but only half of these patients are diagnosed and medicated correctly. Reasons are difficulties in finding the exact diagnosis on the one hand and the vague aetiology and complex effects of antidepressants on the other. The origin of a depressive episode has multiple reasons, especially genetic components are considered to play an important role. Neurobiological investigations of the molecular and biochemical backgrounds of depressive episodes centre on three hypotheses: The dysbalance hypothesis of neurotransmitters (monoamine hypothesis), the hypothesis of activated hypothalamus-pituitary-adrenal axis and the neurotrophin hypothesis. In this context, the hypothesis of activated hypothalamus-pituitary-adrenal axis deals with the dysregulation and over-stimulation of the hypothalamus-pituitary-adrenal axis and the following elevated cortisol-secretion. The monoamine hypothesis of depressive episodes postulates a dysfunction of serotonergic, noradrenergic and dopaminergic systems. The neurotrophin hypothesis refers to the cyclic adenosine monophosphate (cAMP) second messenger system, activated by antidepressants. Cyclic adenosine monophosphate activates the cAMP dependent protein kinase (PKA) that phosphorylates and consequently stimulates the transcription factor cAMP response element binding protein (CREB). Activated CREB enhances the transcription of numerous genes, including the brain derived neurotrophic factor (BDNF) that is among others regulating neurotransmitters and influences neuronal survival, differentiation and plasticity. Astrocytes are part of the macroglia in central nervous system and form the major population of glial cells. They are considered to be responsible for homeostasis of potassium and the regulation of synaptic transmission and neurovascular linkage. Furthermore, astrocytes take part in forming glial cicatrixes, induction of blood-brain-barrier and neuronal survival. In the course of investigating the effects of antidepressants, the centre of interest has focused on astrocytes and their contribution to antidepressant therapy. The aim of this dissertation was to study, if the serotonin transporter (5-HTT), the brain derived neurotrophic factor (BDNF), the dopamin transporter or the nitric oxide synthase III (eNOS) is synthesised in astrocytes and in case of, if their expression is modulated by antidepressants. The astrocyte cultures, gained from rats’ brains, were incubated with the tricyclic antidepressant imipramine, the selective serotonin reuptake-inhibitor escitalopram or sodium chloride solution. After extracting the corresponding mRNA from the astrocyte cultures, it was translated to cDNA and quantified using the quantitative Real-Time PCR. Comparing the expression of the gene of interest with the expression of the housekeeping genes 18s-rRNA, glyceraldehydes phosphate dehydrogenase (GAPDH) and acidic ribosomal phosphoprotein (ARP), equated inexactness of cDNA synthesis and normalised the acquired data. The mathematical evaluation of the qRT-PCR based on the Ct-figures with the assistance of geNORM software. The results show a significantly elevated BDNF expression after imipramine administration. mRNA batches of astrocyte cultures incubated with 100µM imipramine presented more significant results than mRNA batches of astrocyte cultures incubated with 50µM imipramine. Analysing all samples incubated for four hours with the corresponding concentration of imipramine and compared with the sodium chloride solution controls, a significant elevation of BDNF expression is detected in 50µM imipramine samples as well as in 100µM imipramine samples. Escitalopram stimulates the expression of BDNF nominally without showing significant effects. Furthermore, a clear expression of 5-HTT-mRNA in astrocyte cultures could be detected, but none of the antidepressants induced a significant effect on 5-HTT expression. Neither in medicated nor in control samples DAT or NOS-III were detected.

Submitted by PPG Gerontologia Biom?dica (geronbio@pucrs.br) on 2017-11-29T17:04:43Z No. of bitstreams: 1 VEDOVELLI_KELLEM_TESE.pdf: 2587462 bytes, checksum: 3fe069c3be1a03d543e406d18fdf3317 (MD5) / Approved for entry into archive by Caroline Xavier (caroline.xavier@pucrs.br) on 2017-12-04T15:44:11Z (GMT) No. of bitstreams: 1 VEDOVELLI_KELLEM_TESE.pdf: 2587462 bytes, checksum: 3fe069c3be1a03d543e406d18fdf3317 (MD5) / Made available in DSpace on 2017-12-04T15:54:01Z (GMT). No. of bitstreams: 1 VEDOVELLI_KELLEM_TESE.pdf: 2587462 bytes, checksum: 3fe069c3be1a03d543e406d18fdf3317 (MD5) Previous issue date: 2017-08-25 / Coordena??o de Aperfei?oamento de Pessoal de N?vel Superior - CAPES / Introduction: Physical activity has been proposed as a promising intervention to improve cognition and decrease the risk of dementia in older adults. Brain-derived neurotrophic factor (BDNF) seems to mediate, at least partially, these effects of exercise. However, intervention studies of the effects of multimodal exercises on cognition and BDNF levels are scarce and composed by small samples. Thus, the generalization of the conclusions of these studies depends on the reproducibility of the results. Objective: to contribute to the knowledge on the field, the present study evaluated the effects of a physical activity intervention composed by muscle strengthening and aerobic conditioning on BDNF levels and cognition in older women. Methods: Twenty-two independent and non-demented subjects (? 75 years) were assigned to a three-month physical activity intervention (60 min exercise sessions three times a week, 30 min of muscle strengthening and 30 min of aerobic activity) or to a control condition (n= 10, no exercise). The clinical parameters (anxiety and depression symptoms), neuropsychological (Digit Span, Stroop, Trail Making and Contextual Memory tests), physical (upper and lower limb strength, aerobic conditioning) and physiological (serum BDNF) parameters were evaluated immediately before, one and three-months after starting intervention. Results: the control group had stable levels for all measured variables, whereas the intervention group improved on physical fitness, depressive symptoms, cognitive performance and BDNF levels. Moreover, a linear regression identified an association between aerobic conditioning and BDNF levels. Conclusion: The combination of muscle strengthening and aerobic conditioning was able to improve cognitive performance and increase BDNF levels. Aerobic conditioning seems to be an important mediator of these results. / Introdu??o: A atividade f?sica tem sido proposta como uma interven??o promissora para melhorar a cogni??o e diminuir o risco de dem?ncia em adultos mais velhos. O fator neurotr?fico derivado do c?rebro (BDNF) parece mediar, pelo menos parcialmente, esses efeitos do exerc?cio. Entretanto, os estudos de interven??o dos efeitos de exerc?cios multimodais sobre o desempenho cognitivo e os n?veis de BDNF s?o escassos e compostos por pequenas amostras. Assim, a generaliza??o das conclus?es destes estudos depende da reprodutibilidade dos resultados. Objetivo: avaliar os efeitos de uma interven??o de atividade f?sica composta pelo fortalecimento muscular e condicionamento aer?bico nos n?veis de BDNF e cogni??o em mulheres idosas, de modo a contribuir para o avan?o cient?fico na ?rea. M?todos: Idosas independentes e n?o demenciadas (? 75 anos) foram submetidas a uma interven??o de tr?s meses de atividade f?sica (n = 22, sess?es de exerc?cios de 60 min tr?s vezes por semana, sendo 30 min de fortalecimento muscular e 30 min de atividade aer?bia) ou a uma condi??o de controle (n = 10, sem exerc?cio). Foram avaliados os par?metros cl?nicos (sintomas de ansiedade e depress?o), neuropsicol?gicos (testes de Span de D?gitos, Stroop, Trail Making e Mem?ria Contextual), f?sicos (for?a dos membros superiores e inferiores, condicionamento aer?bico) e fisiol?gicos (BDNF s?rico) no in?cio do protocolo, 1 e 3 meses ap?s o in?cio da interven??o. Resultados: O grupo controle apresentou n?veis est?veis ao longo do tempo para todas as vari?veis medidas, ao passo que o grupo de interven??o melhorou a aptid?o f?sica, sintomas depressivos, desempenho cognitivo e n?veis de BDNF. Al?m disso, uma regress?o linear identificou uma associa??o entre o condicionamento aer?bio e os n?veis de BDNF. Conclus?o: A combina??o de fortalecimento muscular e condicionamento aer?bio foi capaz de melhorar o desempenho cognitivo e aumentar os n?veis de BDNF. O condicionamento aer?bio parece ser um importante mediador desses resultados.

Introdução: A formação da memória envolve mudanças na expressão de genes neuronais. Remodelações epigenéticas da cromatina e modificações pós-traducionais reversíveis no DNA ou nas proteínas histonas representam mecanismos centrais na regulação da expressão gênica durante o desenvolvimento do cérebro e a aprendizagem inicial ou recuperação da memória. Desequilíbrios nos níveis de acetilação de histonas estão associados à uma ampla variedade de desordens cerebrais. Histonas deacetilases (HDACs) desempenham um papel fundamental na homeostase da acetilação de histonas e na regulação de atividades celulares fundamentais como a transcrição, tornando-as um foco de estudo. Evidências mostram que a administração de inibidores de histonas deacetilases (HDACis) restauram a memória associada à regulação da expressão gênica e melhora a memória em ratos. Estudos em modelos animais têm mostrado que a formação da memória envolve uma série de alterações bioquímicas em várias áreas do sistema nervoso central, entre as quais se destacam o hipocampo e a amígdala basolateral (BLA). Neste contexto, fármacos experimentais, como a tricostatina A (TSA), que atuam sobre mecanismos epigenéticos, têm sido recentemente propostos como potenciais terapias para o tratamento de disfunção cognitiva e memória associados a doenças neurológicas e psiquiátricas. Objetivo: Neste trabalho objetivamos compreender e elucidar o papel da acetilação de histonas em processos envolvidos na modulação da memória utilizando o fármaco TSA e se baseia na hipótese de que a atividade de HDACs é essencial para a modulação das respostas de aprendizado na tarefa de esquiva inibitória (IA). Métodos: Ratos Wistar foram canulados bilateralmente na amígdala. Os efeitos das micro-infusões intra-amigdalares de TSA foram observados na consolidação e na extinção da memória após o treino na tarefa de esquiva inibitória e nos níveis do fator neurotrófico derivado do cérebro (BDNF) na BLA e no hipocampo referentes à consolidação da memória. Resultados: Os resultados demonstraram que a infusão intra-amigdalar de TSA 1.5 h, 3 h e 6 h após o treino na tarefa de esquiva inibitória resulta na melhora da memória de longa duração (LTM). TSA acelerou a extinção da memória quando infundido imediatamente pós-teste. Além disso, aumentou os níveis de BDNF no hipocampo. Conclusão: Estes resultados indicam que eventos epigenéticos possuem um papel importante no aprendizado e na memória através da atividade de HDACs. / Introduction: Memory formation involves changes in the expression of neuronal genes. Epigenetic remodeling of chromatin and reversible post-translational modifications in the DNA or in the histone proteins represent central mechanisms in the regulation of gene expression during brain development and early learning or memory retrieval. Imbalances in the levels of histone acetylation are associated with a wide variety of brain disorders. Histone deacetylases (HDACs) play a key role in homeostasis of histone acetylation and regulation of fundamental cellular activities, such as transcription, making them a focus of study. Evidences shows that the administration of histone deacetylases inhibitors (HDACis) restore the memory associated with the regulation of gene expression and improves memory in rats. Studies in animal models have shown that memory formation involves a series of biochemical changes in several areas of the central nervous system, which the hippocampus and basolateral amygdala (BLA) are the most highlighted. In this context, experimental drugs, such as trichostatin A (TSA), that act on epigenetic mechanisms, have recently been proposed as potential therapies for the treatment of memory and cognitive dysfunction associated with psychiatric and neurological disorders. Objective: In this work we aimed to understand and elucidate the role of histone acetylation in processes involved in memory modulation using the drug TSA and is based on the hypothesis that HDACs activity is essential for the modulation of learning answers in the inhibitory avoidance (IA) task. Methods: Wistar rats were cannulated bilaterally in the amygdala. The effects of TSA micro-infusions into the BLA were observed in the consolidation and extinction of memory after training in the inhibitory avoidance task and the levels of brain-derived neurotrophic factor (BDNF) in the BLA and hippocampus related to memory consolidation. Results: The results demonstrated that the TSA infusion into BLA 1.5 h , 3 h and 6 h posttraining in the inhibitory avoidance task results in improved long-term memory (LTM). TSA accelerated the extinction of memory when infused immediately post-test. In addition, increased levels of BDNF in the hippocampus. Conclusion: These results indicate that epigenetic events play an important role in learning and memory by HDAC activity.

Existem crescentes evidências sobre a contribuição da liberação de noradrenalina (NA) central na consolidação das memórias. Teoricamente, o Núcleo do Trato Solitário (NTS) recebe informações e diversos estímulos periféricos, que são então projetados ao Núcleo Paragigantocelular (PGi). Este, por sua vez, utiliza neurotransmissores, predominantemente excitatórios, para influenciar a ativação do Locus Coeruleus (LC). Então, o LC envia projeções noradrenérgicas ao hipocampo e à amígdala, influenciando os processos mnemônicos. Aqui nós demonstramos que a inibição pelo muscimol do NTS, PGi ou LC até 3 horas após o treino na tarefa de reconhecimento de objetos (RO) impede a consolidação da memória medida 24 h após o treino. Adicionalmente, a infusão de timolol, um antagonista de receptores β-adrenérgicos, na região CA1 do hipocampo também impede a consolidação deste tipo de memória. A infusão de NA na região CA1 do hipocampo não altera a retenção da memória, mas, reverte o prejuízo causado pela inibição do NTS, PGi ou LC. A infusão de NMDA no LC após a inibição do NTS ou PGi também reverte essa amnésia. Concomitantemente, verificamos que a inibição NTS, PGi ou LC bloqueia o aumento da expressão do fator neurotrófico derivado do cérebro (BDNF, do inglês brain-derived neurotrophic factor) que ocorre 120 min após o treino na tarefa de reconhecimento de objetos na região CA1 do hipocampo. Também a infusão de NA na região CA1 do hipocampo após a inibição do NTS, PGi ou LC ou de NMDA no LC após a inibição do NTS ou PGi promovem novamente o aumento do BDNF120 min após o treino no RO. Com isso conclui-se que a ativação da via NTS-PGi-LC-Hipocampo é necessária para que ocorra consolidação da memória de RO, na qual desempenha um papel o BDNF hipocampal. / There is evidence of the contribution of brain noradrenaline release (NA) to memory consolidation. The Nucleus of the Solitary Tract (NTS) receives information originated by peripheral stimuli and projects to the Paragigantocellularis Nucleus (PGi), which influences the Locus Coeruleus (LC) through excitatory neurotransmitters. The LC sends noradrenergic projections to the hippocampus and amygdala, influencing the memory processes. Here we show that inhibition by muscimol of NTS, PGi or LC up to 3 h after object recognition training impairs the consolidation of the memory measured 24 h later. Additionally, the infusion of timolol in the CA1 region of hippocampus also inhibits consolidation of this type of memory. The infusion of NA into the CA1 region of hippocampus does not alter memory consolidation of this task, but reverts the deleterious effect of NTS, PGi or LC inhibition. The infusion of NMDA in LC after inhibition of NTS or PGi also reverts the amnesia. Concomitantly, the inhibition of NTS, PGi or LC blocks the increase of brain-derived neurotrophic factor (BDNF) expression in CA1 that occurs 120 min after training in the object recognition task. Further, the infusion of NA in CA1 after inhibition of NTS, PGi or LC; or of NMDA in LC after inhibition of NTS or PGi promotes the BDNF increase seen 120 min after object recognition training. Thus, it is concluded that the activation of NTSPGi- LC-Hippocampus pathway is necessary for consolidation of the object recognition memory, and hippocampal BDNF is involved in this process.

O consumo de cafeína tornou-se popular em adolescentes devido ao aumento da ingestão de bebidas comercializadas na forma de bebidas energéticas. Alguns estudos consideram que os efeitos benéficos da cafeína são atribuídos a uma reversão dos sintomas de abstinência. Neste estudo, foram investigados os efeitos da administração crônica de cafeína em ratos desde a adolescência (40 dias de idade) até à idade adulta (3 meses de idade) na memória de reconhecimento e BDNF e proteínas relacionadas a sua sinalização nas regiões do córtex e hipocampo. A cafeína (0,3 e 1,0 g/L) foi administrada na água de beber durante o ciclo de ativo dos animais (ciclo escuro) e retirada durante os fins de semana. Este protocolo foi desenvolvido a fim de mimetizar o consumo humano. Para as experiências de privação (abstinência),a administração crônica foi interrompida 24 ou 48 h antes do teste de tarefa de reconhecimento de objetos. Na tarefa de reconhecimento de objetos, foi possível observar os efeitos positivos da cafeína sobre a memória de reconhecimento, pois os animais tiveram um bom desempenho na tarefa. Entretanto, mesmo após a interrupção do tratamento os animais continuaram desempenhando bem a tarefa, dessa forma a abstinência de um tratamento crônico com cafeína não influencia a memória de reconhecimento. A cafeína na sua dose mais alta (1,0 mg / mL) e 24 h após a retirada, causou uma diminuição nos níveis de BDNF no hipocampo e nenhum efeito sobre as proteínas proBDNF e TrkB. Em contrapartida, no córtex a cafeína em ambas as doses diminuui os níveis de BDNF, um efeito que persistiu apenas para a dose mais elevada em ambos os tempos de retirada. O ProBDNF teve seus níveis diminuídos pela cafeína (1,0 mg / mL) após 48 horas da retirada, enquanto a cafeína em ambos os tempos de retirada aumentou receptores TrkB. Como mencionado anteriormente, estes resultados mostraram que a cafeína administrada durante a adolescência até a idade adulta, seguida da sua retirada não afeta a memória de reconhecimento. Estes efeitos poderiam ser atribuídos ao desenvolvimento da tolerância por tratamento crónico. Além disso, o aumento de receptores de TrkB seguido por diminuição de BDNF pode ser contribuído para a ausência de efeitos de abstinência de cafeína na memória de reconhecimento. / The caffeine consumption has become popular among adolescents due to increased intake of beverages marketed as energy drinks. Some studies consider that the beneficial effects of caffeine are attributable to a reversal of withdrawal symptoms. This study investigated the effects cronic administration caffeine in rats since adolescent period (40 days old) until adulthood (3 months old) on recognition memory and BDNF-related proteins and their signalling in the regions of the cortex and hipocampus. Caffeine (0.3 and 1.0 g/L) was administered in drinking water during the light cycle and discontinued at weekends. This protocol was developed to mimic human consumption. For withdrawal experiments, cronic administration of caffeine was interrupted 24 or 48 h before the test session on object recognition task. In the task, we observed the positive effects of caffeine on recognition memory once that animals learned the task. However, even after treatment interuption animals continued performing the task, so the withdrawal of chronic treatment with caffeine has no effect on recognition memory. Caffeine at its highest dose (1.0 mg / mL) after 24 h and after removal, showed a decrease in BDNF levels in the hippocampus and no effect on protein proBDNF and its receptor TrkB. In contrast, in the cortex caffeine decreased BDNF levels at both doses, an effect which persisted for only the highest dose at both time of withdrawal. ProBDNF levels had decreased by caffeine (1.0 mg / mL) after 48 hours of removal, while the caffeine in both periods of increased withdrawal TrkB receptors. As mentioned earlier, these results showed that caffeine administered during adolescence to adulthood, followed by its removal does not affect recognition memory. These effects could be attributed to the development of tolerance for chronic treatment. Furthermore, the increase of TrkB receptors followed by a decrease in BDNF may be contributed to the absence of withdrawal effects of caffeine in recognition memory.

Introdução: A exposição crônica a diferentes tipos de dieta altera o metabolismo hipotalâmico e mesolímbico, podendo causar alterações no comportamento alimentar do indivíduo. O BDNF, fator de crescimento neuronal, pode atuar na modulação do comportamento alimentar tanto em vias hedônicas quanto homeostáticas. O objetivo do estudo foi investigar como o BDNF atua na modulação do comportamento alimentar em vias homeostáticas e hedônicas em ratas fêmeas com diferentes perfis metabólicos. Materiais e métodos: Ratas Wistar fêmeas adultas randomizadas por peso foram divididas em: dieta controle (C) contendo 22% de proteína e 4% de lipídios; dieta hipoproteica (LP) 8% de proteína ou dieta hiperlipídica (HF) 45% de lipídios, ad libitum, por 5 semanas, sendo o consumo medido a cada 72 horas e o peso semanalmente. O trabalho foi dividido em duas partes. Na primeira parte, após as 5 semanas de dieta os animais ficaram em jejum por 4 horas e foram expostos ao alimento doce (Froot Loops®), previamente pesado, por 1 hora, a fim de verificar o consumo de alimento palatável em ratas com diferentes perfis metabólicos. Imediatamente após coletou-se sangue e cérebro, assim como, o peso da gordura abdominal foi mensurado. Na segunda parte do estudo, após as 5 semanas de dieta, os animais ficaram durante 7 dias no BioDAQ®, um sistema computadorizado de análise do comportamento alimentar, para avaliar o consumo da dieta habitual em ratas com diferentes perfis metabólicos. O consumo foi mensurado através de mordidas (diferença de 0,1 g na balança) e refeições (conjunto de porções por um tempo igual ou menor a 15 min, porções são mordidas ininterruptas). No dia 10 foi realizado o teste de preferência alimentar no qual o animal poderia escolher entre a dieta habitual (dieta que eles receberam por 5 semanas) ou a dieta hipersacarídica (HP) (contendo 34% lipídios, 30,2% carboidratos, 14% proteínas, 20% sacarose), com duração de 20 horas. Após 1 semana, foi coletado sangue, cérebro e mensurada a gordura abdominal. Foi realizado western blotting para tirosina hidroxilase (TH) e fosfo- tirosina hidroxilase (pTH) no núcleo accumbens, STAT3 e fosfo-STAT3 (pSTAT3) no hipotálamo. Adicionalmente, foi mensurado BDNF no soro, no núcleo do trato solitário (NTS), área tegmentar ventral (VTA) e glicemia no soro. Resultados: Nas 5 semanas de tratamento, em relação ao ganho de peso dos animais não houve diferenças significativas entre os grupos, não houve interação, apenas apresentaram efeito do tempo (p< 0,001). A gordura abdominal foi maior nas ratas HF (p= 0,002) e LP (p= 0,023) em relação aos controles. Os animais que receberam dieta HF comeram menos gramas de Froot Loops® do que os animais controle (p= 0,003). Durante a habituação ao BioDAQ® não houve diferença significativa no consumo de dieta entre os grupos. Na análise do comportamento alimentar no BioDAQ®, comparando os grupos controle versus hiperlídica (C x HF), animais HF tiveram o tamanho da refeição (g) (p=0,049), número de porções (p= p< 0,001), tamanho da refeição ciclo escuro (p= 0,002), número de porções ciclo escuro (p < 0,001) menor do que os controles e tamanho da porção ciclo escuro (p=0,006) maior do que os controles. Na análise do comportamento alimentar, comparando os grupos controle versus hipoproteica (C x LP) foram encontradas diferenças significativas nos seguintes parâmetros: média do tamanho da porção (g) (p= 0,004), média do tamanho da porção no ciclo claro (g) (p= 0,042), média do tamanho da refeição no ciclo escuro (g) (p= 0,035), média do tamanho da porção no ciclo escuro (g) (p= 0,006). Em todos os parâmetros os animais LP tiveram uma média maior do que os animais controle. No teste de preferência alimentar, os animais HF apresentaram uma média inferior aos animais controle nos seguintes parâmetros: consumo de dieta hipersacarídica (Kcal) (p= 0,034), número de refeições de dieta hipersacarídica (p= 0,016), número de porções de dieta hipersacarídica (p= 0,001). Os animais HF apresentaram médias superiores aos animais C em relação ao tamanho da porção de dieta hipersacarídica (g) (p= 0,023), PMI (intervalo entre refeições) total de dieta hipersacarídica (p= 0,022), saciedade total de dieta hipersacarídica (p= 0,008). Durante o teste de preferência alimentar comparando-se controle versus hipoproteica (C x LP), os animais (LP) apresentaram o consumo de dieta hipersacarídica (kcal) (p= 0,030) e o número de porções de dieta hipersacarídica (p= 0,003) menor que os animais controles. Os animais LP apresentaram saciedade total de dieta hipersacarídica (p= 0,046) maior do que os controles. Não houve diferenças significativas nos níveis de glicemia no soro entre os grupos. Os animais HF apresentaram as médias de pSTAT3 maior do que os controles (p= 0,013) e fosfo-tirosina hidroxilase menor do que os controles (p= 0,05). Os animais LP apresentaram STAT3 menor do que os animais controle (p= 0,035) e tiveram resultados de BDNF próximos da significância (p=0,053), tendo apresentado uma média inferior aos animais controle. Conclusões: A exposição aos diferentes tipos de dieta muda os padrões de alimentação bem como a estrutura corpórea. Tanto animais expostos a dieta HF quanto a LP apresentam alterações compatíveis com um estado de pré-resistência à leptina. O BDNF parece modular as vias homeostáticas e hedônicas do comportamento alimentar, no entanto mais estudos são necessários para entendermos melhor esses mecanismos. / Introduction: The chronic exposure to different types of diet modifies the hypothalamic and mesolimbic metabolism, what can lead to changes in the individual feeding behavior. BDNF, a neuronal growing factor, can act modulating feeding behavior in hedonic as well as in homeostatic pathways. The aim of this study was to investigate how BDNF acts modulating feeding behavior regarding hedonic and homeostatic pathways in female rats with different metabolic profiles. Materials and methods: Female adult Wistar rats randomized by weight were divided in: control diet (C) with 22% of protein and 4% of lipids; low-protein diet (LP) with 8% of protein or high fat diet (HF) with 45% of lipids, ad libitum, for 5 weeks, with the diet consumption verified every 72 hours and the weight verified weekly. The study was divided in two parts. In the first part, after 5 weeks receiving diet, animals were submitted to 4 hours of fasting and then were exposed to sweet food (Froot Loops®), previously weighted, for 1 hour to verify the palatable food consumption in rats with different metabolic profiles. Immediately after, blood and brain were collected and the abdominal fat weight was verified. In the second part of the study, after 5 weeks receiving diet, the animals were submitted to the BioDAQÒ, a computer system of feeding behavior analyses, to evaluate the habitual diet consumption in rats with different metabolic profiles for 7 days. The consumption was measured through bites (0.1g of difference in the scale) and meals (a group of portions for a period of time less or equal than 15min, portions are uninterrupted bites). On day 10 the food preference test was done and the animals could choose between the habitual diet (the one they have received for 5 weeks) and the high sucrose palatable diet (HP) (with 34% of lipids, 30.2% of carbohydrates, 14% of protein, 20% of sucrose) for a period of 20 hours. After one week, blood and brain were collected and abdominal fat was measured. Western blotting was used to verify the content of tyrosine hydroxylase (TH) and phospho-tyrosine hydroxylase (pTH) in the nucleus accumbens, STAT3 and phospho-STAT3 (pSTAT3) in the hypothalamus. Additionally, BDNF was measured in the serum, in the solitary tract nucleus (NTS) and in the ventral tegmental area (VTA) and glycemia was also verified. Results: Considering the 5 weeks of treatment regarding animals weight gain there was no significant difference between groups and no interaction, only an effect of time (p<0.001). Abdominal fat was higher in HF (p=0.002) and LP (p=0.023) rats comparing to controls. Animals that received HF diet ate less grams of Froot Loopsâ than controls (p=0.003). During the habituation to the BioDAQÒ no significant difference was seen in the consumption of the diet between the groups. When analyzing feeding behavior in the BioDAQÒ, comparing control against high fat groups (C x HF), HF animals had a decrease in the size of the meal (g) (p=0.049), number of portion (p<0.001), meal size in the dark phase (p=0.002), number of portions in the dark phase (p<0.001) compared to controls while the size of the portion in the dark phase (p=0.006) was bigger than controls. Analyzing feeding behavior comparing control against low-protein groups (C x LP) a significant difference was observed in the following parameters: average of the size of the portion (g) (p=0.004), average of the size of the portion in the light phase (g) (p=0.042), average of the size of the meal in the dark phase (g) (p=0.035), average of the size of the portion in the dark phase (g) (p=0.006). LP animals showed a higher average than controls in all parameters. In the food preference test, HF animals showed a lower average than controls in the following parameters: consumption of high sucrose palatable diet (Kcal) (p=0.034), number of meals of high sucrose palatable diet (p=0.016), number of portions of high sucrose palatable diet (p=0.001). HF animals showed higher averages than controls regarding the size of the portion of the high sucrose palatable diet (g) (p=0.023), total PMI (post interval meal) of high sucrose palatable diet (p=0.022) and total satiety of high sucrose palatable diet (p=0.008). In the food preference test comparing controls against low-protein groups (C x LP), LP animals showed a lower consumption of high sucrose palatable diet (Kcal) (p=0.030) and a lower number of portions of high sucrose palatable diet (p=0.003) than control animals. LP animals showed total satiety of high sucrose palatable diet (p=0.046) bigger than controls. There was no significant difference in the glycemia levels between groups. HF animals had pSTAT3 averages higher than controls (p=0.013) and phospho-tyrosine hydroxylase lower than controls (p=0.05). LP animals showed STAT3 lower than control animals (p=0.035) and had BDNF results close to the significance (p=0.053), showing a lower average than control animals. Conclusion: The exposure to different types of diets changes the feeding patterns as well as the body structure. Animals exposed to HF diet as well as those exposed to LP diet showed changes related to a leptin pre-resistant state. BDNF seems to modulate homeostatic and hedonic feeding behavior pathways, however more studies are necessary to a better understanding of the mechanisms.

Submitted by Cristiane Chim (cristiane.chim@ucpel.edu.br) on 2017-11-13T11:16:03Z No. of bitstreams: 1 DANIELA GOMEZ MARTIN ISERHARDT.pdf: 1551026 bytes, checksum: d7fce1f77757f051d25d76f86cfbb6b8 (MD5) / Made available in DSpace on 2017-11-13T11:16:03Z (GMT). No. of bitstreams: 1 DANIELA GOMEZ MARTIN ISERHARDT.pdf: 1551026 bytes, checksum: d7fce1f77757f051d25d76f86cfbb6b8 (MD5) Previous issue date: 2016-07-26 / Fibromyalgia is a rheumatic syndrome causing musculoskeletal pain diffuse and chronic. Aerobic exercises in the pool can cause changes in Brain derived neurotrophic factor (BDNF) causing analgesia in these patients. Muscle upper trapezius surface electromyography assists in the treatment conduct and pain points in the shoulder girdle being inhibited, promote analgesia for patients. This work aimed to analyze the changes in BDNF levels in women with fibromyalgia after practice a short protocol of hydrotherapy. Secondary objectives were to evaluate the effect of hydrotherapy in symptoms of pain and in quality of life of women with fibromyalgia and analyze the relationship between water exercise with muscle activations of upper trapezius electromyography. Case-control study with 13 women diagnosed with fibromyalgia in a case group and the control group 11 women without fibromyalgia. The participants were assessed through questionnaires and undergo a short hydrotherapy protocol; a total of 10 sessions of 60 minutes twice a week for five weeks. A statistically significant increase in BDNF levels was observed in fibromyalgia patients between pre and post tenth session. Regarding the pain, there was a statistically significant decrease in pre and post hydrotherapy in both groups. Muscle activations of the upper trapezius showed a significant decrease, but not related when compared to pain. We conclude that BDNF may have altered in a short moderate aerobic protocol of exercises when measured and analyzed in a chronic way. Electromyography showed that the fibromyalgia patients had disturbance of the autonomic nervous system. The study demonstrated that the heated water, along with its physical properties and heart rate zone worked could change pain parameters and improve the quality of life state of patients with fibromyalgia. / A fibromialgia é uma síndrome reumática ocasionando dor músculoesquelética difusa e crônica. Exercícios aeróbicos na piscina podem provocar mudanças no Brain derived neurotrophic factor (BDNF) provocando analgesia nesses pacientes. A eletromiografia de superfície do músculo trapézio superior auxilia na conduta de tratamento e os pontos de dor na cintura escapular estando inibidos, promovem analgesia para as pacientes. Esse trabalho teve como objetivo principal analisar as alterações nos níveis de BDNF em mulheres com fibromialgia após a prática de um protocolo de curta duração de hidroterapia. Os objetivos secundários foram: avaliar o efeito da hidroterapia, na sintomatologia de dor e na qualidade de vida de mulheres com fibromialgia e analisar a relação entre exercícios na água com as ativações musculares via eletromiografia do trapezio superior. Estudo tipo caso controle com 13 mulheres diagnósticadas com fibromialgia no grupo dos casos e no grupo controle 11 mulheres sem fibromialgia. As participantes foram avaliadas através de questionários e submetidas a um curto protocolo de hidroterapia; totalizando 10 sessões de 60 minutos, duas vezes por semana, durante cinco semanas. Um aumento estatisticamente significativo nos valores de BDNF foram observados nas pacientes com fibromialgia entre a pré e pós décima sessão. Com relação a dor, houve diminuição estatisticamente significativa pré e pós a hidroterapia nos dois grupos. As ativações musculares do trapézio superior mostraram uma diminuição significativa, porém quando comparado com dor não apresentou relação. Concluímos que o BDNF pode se apresentar alterado em um protocolo de exercicios de curta duração aeróbico moderado quando mensurado e analisado de maneira crônica. A eletromiografia demonstrou que as pacientes com fibromialgia apresentaram distúrbio do sistema nervoso autônomo. O estudo pôde demonstrar que a água aquecida, juntamente com suas propriedades físicas e a zona de frequência cardíaca trabalhada puderam alterar parâmetros de dor e melhorar o estado da qualidade de vida das pacientes com fibromialgia.

Submitted by Cristiane Chim (cristiane.chim@ucpel.edu.br) on 2017-11-27T11:42:50Z No. of bitstreams: 1 FERNANDA BURLANI NEVES.pdf: 1796364 bytes, checksum: 1cd868160548173be0f57cea7319fb27 (MD5) / Made available in DSpace on 2017-11-27T11:42:50Z (GMT). No. of bitstreams: 1 FERNANDA BURLANI NEVES.pdf: 1796364 bytes, checksum: 1cd868160548173be0f57cea7319fb27 (MD5) Previous issue date: 2015-02-27 / The purpose of this study was to contribute to the knowledege of women’s health in perimenopause phase and analyse the symptons related to this period with women’s quality of life. This period is characterized by a critical transition where several disturbances occurs due to endocrinous, psichological and biological alterations. This alterations generally are marked by periods of depression, anxiety and hormone levels alterations, particularly on strogens levels. Firstly it was conducted a bibliographic research with the objective of verify the effect of physical activity (PA) on psichological symptons of menopause period. After that, a transversal study was realized to evaluate BDNF, NGF e GDNF levels and the relation of these seric proteins levels with quality of life in climateric women. It was applied a questionnaire to identify socioeconomic, demographic and health behavior matters of these women. To evaluate quality of life it was administered Women’s Health Questionnaire (WHQ) and blood sample was colected from all sample. Statistical analyses were conducted using SPSS 21 and GraphPad Prism 6.0. Non-parametric test was performed and all values were presented as mean ± standart deviation. Spearman coeficent was used to analyse correlation between BDNF, GDNF, NGF and classifications of WHQ. p < 0,05 was cosidered significant. About serum NGF, the association between neurotrophic factor and WHQ presented a correlation between NGF and anxiety domain (r = 0,373, p ≤ 0,05) and attractiveness domain (r = 0,344, p ≤ 0,05). The literature review pointed that PA could promote reduction of stress and depressive symptons and better quality of life. Correlations between seric levels of BDNF, GDNF, NGF reveals that women who presented higher scores in anxiety and attractiveness domains had higher seric level of NGF in comparison with those who presented lower scores in these refered domains. Results suggests that this neutrophin could perform a relevant function related to behaviour during perimenopause period. Thus, more studies are needed to elucidate this question with te aim of reach clearly understanding of facts related to this period. The perspectives of this study are evaluate psichological and hormonal aspects by the effect of anaerobic ans aerobic training in cronic and acute phases, analyse cognitive function, control hormonal replacement, increase the number of participants and idealize PA as non-pharmacological treatment in perimenopause and all phases of women’s life. / Este estudo tem como objetivo contribuir para o conhecimento da saúde das mulheres na fase da perimenopausa e ainda analisar os sintomas referentes a esse período na qualidade de vida dessas mulheres. Esse período é caracterizado por uma crítica transição na qual a mulher lida com diversos transtornos consequentes de alterações endócrinas, psicológicas e biológicas que variam de acordo com as características da personalidade de cada mulher. As alterações geralmente são marcadas por períodos de depressão, ansiedade e alterações nos níveis hormonais, particularmente nos níveis de estrogênio. Inicialmente realizou-se uma pesquisa Bibliográfica com objetivo de verificar o efeito da Atividade Física (AF) nos sintomas psicológicos no período da menopausa, após desenvolveu-se um estudo transversal com objetivo de avaliar os níveis de BDNF, NGF e GDNF em mulheres climatéricas e a relação desses níveis séricos destas proteínas com a qualidade de vida de mulheres que estão passando por essa fase vital. Primeiramente foi aplicado um instrumento para identificar o nível socioeconômico e condições demográficas e questões de saúde comportamental. Para determinar a qualidade de vida foi usado Women’s Health Questionnaire (WHQ) e foi coletado amostra sanguínea de todas as participantes. Para as análises estatísticas foram realizadas utilizando SPSS 21 e GraphPad Prism 6.0. Foi realizado teste não paramétrico e todos os valores foram apresentados como média ± desvio padrão. O Coeficiente de correlação de Spearman foi utilizado para analisar a correlação entre BDNF, GDNF, NGF e as pontuações do WHQ. Os valores de p < 0,05 foram considerados estatisticamente significativos. Em relação aos níveis de NGF no soro, a associação entre fator neurotrófico e os domínios WHQ apresentou uma correlação entre os níveis de NGF com domínio ansiedade (r = 0,373, p ≤ 0,05) e domínio de atratividade (r = 0,344, p ≤ 0,05). Foi possível verificar na revisão de literatura que a AF pode promover a redução da ansiedade, do estresse e dos sintomas depressivos e a melhora da qualidade de vida. Na correlação entre os níveis séricos do BDNF, GDNF, NGF com o WHQ foi observado que as mulheres com escores mais altos em domínios de ansiedade e atratividade apresentam o maior nível sérico de NGF em comparação com aquelas com pontuações mais baixas nestes referidos domínios. Estes resultados sugerem que esta neurotrofina pode desempenhar um papel relevante na comportamento durante a perimenopausa. Deste modo, mais pesquisas são necessárias para elucidar essa questão com a intenção de alcançar claramente a compreensão dos fatos envolvidos neste período. Como perspectivas deste estudo pode-se promover a avaliação dos objetivos estudados sob o efeito de um treinamento físico aeróbio e anaeróbio na fase aguda e crônica, avaliar a função cognitiva, controlar o uso da reposição hormonal, aumentar o número das participantes e ainda realizar nas diferentes fases da vida, para de fato propor a Atividade Física como um plano de tratamento efetivo não farmacológico na perimenopausa e em outras fases da vida da mulher.