The goal of my thesis was to elucidate the impact of protein-energy malnutrition (PEM, a condition commonly found in aging stroke patients) on outcomes from global ischemia. I first examined the hypothesis that PEM will impair working memory in the adult gerbil as measured in the T-maze. Gerbils were fed an adequate (12.5%) or low protein (2%; PEM) diet for 6wk. Stringent assessment of T-maze performance indicated an improvement with PEM although I was unable to reconcile whether this was increased motivation for the food reward or enhanced working memory.<p> The second hypothesis tested was PEM will decrease expression of plasticity-associated hippocampal mRNA and protein expression following global ischemia in the gerbil. The plasticity markers brain-derived neurotrophic factor (BDNF), tropomyosin-related kinase B (trkB), and growth-associated protein-43 (GAP-43) were examined in the CA1 hippocampal region post-ischemia. PEM induced in gerbils for 4wk did not alter the global ischemia-induced decrease in CA1 neurons. Ischemia resulted in increased CA1 pyramidal expression of BDNF and trkB mRNA at 1, 3, and 7d post-ischemia and increased trkB protein expression at 3 and 7d. PEM further elevated the increased trkB protein detected at 7d in the fibres. Ischemia resulted in increased GAP-43 protein at 3 and 7d post-ischemia with PEM increasing this expression at 3d in the CA3 and hilar regions in addition to CA1. These findings suggest an increased stress-response and/or hyperexcitability state in the hippocampus of malnourished ischemic animals.<p> Since the reliability of the gerbil model of global ischemia has come into question, the third part of my thesis tested the hypothesis that the influence of pre-existing PEM on global ischemia-induced hippocampal injury can be reliably studied with the 2-vessel occlusion rat model. The impact of PEM on CA1 neuronal death and dendritic damage was examined. Rats received protein adequate (18%) or deficient (2%; PEM) diet for 7-8d prior to global ischemia. PEM did not worsen the decrease in CA1 neurons and dendrites observed at 7d post-ischemia. Importantly, I found that PEM altered blood glucose and acid-base balance during surgery and caused brief hypothermia post-surgically, factors which are important for consistent brain injury.<p> Taken together, these findings reveal (i) that nutritional care, although frequently ignored, can have robust effects on recovery mechanisms after brain ischemia; and (ii) the challenges of studying pre-existing PEM in an established rodent model of stroke.

The goal of my thesis was to elucidate the impact of protein-energy malnutrition (PEM, a condition commonly found in aging stroke patients) on outcomes from global ischemia. I first examined the hypothesis that PEM will impair working memory in the adult gerbil as measured in the T-maze. Gerbils were fed an adequate (12.5%) or low protein (2%; PEM) diet for 6wk. Stringent assessment of T-maze performance indicated an improvement with PEM although I was unable to reconcile whether this was increased motivation for the food reward or enhanced working memory.<p> The second hypothesis tested was PEM will decrease expression of plasticity-associated hippocampal mRNA and protein expression following global ischemia in the gerbil. The plasticity markers brain-derived neurotrophic factor (BDNF), tropomyosin-related kinase B (trkB), and growth-associated protein-43 (GAP-43) were examined in the CA1 hippocampal region post-ischemia. PEM induced in gerbils for 4wk did not alter the global ischemia-induced decrease in CA1 neurons. Ischemia resulted in increased CA1 pyramidal expression of BDNF and trkB mRNA at 1, 3, and 7d post-ischemia and increased trkB protein expression at 3 and 7d. PEM further elevated the increased trkB protein detected at 7d in the fibres. Ischemia resulted in increased GAP-43 protein at 3 and 7d post-ischemia with PEM increasing this expression at 3d in the CA3 and hilar regions in addition to CA1. These findings suggest an increased stress-response and/or hyperexcitability state in the hippocampus of malnourished ischemic animals.<p> Since the reliability of the gerbil model of global ischemia has come into question, the third part of my thesis tested the hypothesis that the influence of pre-existing PEM on global ischemia-induced hippocampal injury can be reliably studied with the 2-vessel occlusion rat model. The impact of PEM on CA1 neuronal death and dendritic damage was examined. Rats received protein adequate (18%) or deficient (2%; PEM) diet for 7-8d prior to global ischemia. PEM did not worsen the decrease in CA1 neurons and dendrites observed at 7d post-ischemia. Importantly, I found that PEM altered blood glucose and acid-base balance during surgery and caused brief hypothermia post-surgically, factors which are important for consistent brain injury.<p> Taken together, these findings reveal (i) that nutritional care, although frequently ignored, can have robust effects on recovery mechanisms after brain ischemia; and (ii) the challenges of studying pre-existing PEM in an established rodent model of stroke.

Cognitive rehabilitation has been shown to have beneficial effects on functional recovery following traumatic brain injury. In the present study, the rehabilitative effects of cognitive training in the T-maze on functional recovery of behavior and cortical sparing following a cortical impact injury (CCI) were examined. T-maze alternation has a widespread application in detecting cognitive dysfunction, and alternation in particular utilizes working memory. 47 male Sprague-Dawley rats were divided into six groups (sham trained, sham yoked, sham control, injured trained, injured yoked, injured control). Injured animals received a bilateral frontal craniotomy (1.0 A/P, 0.0 M/L from Bregma). The cortices were depressed at a depth of 2.5 mm at a velocity of 3 m/s. T-maze training began on post surgery day 2 and continued daily through post surgery day 19. Following this rehabilitative T-maze training, cognition was assessed using two different memory tasks in the Morris water maze (MWM).

Tem sido proposto que vias serotonérgicas distintas originárias do núcleo dorsal da rafe (NDR) modulariam diferentes tipos de reações de defesa a estímulos aversivos. Deakin e Graeff (1991) propuseram que a ativação da via ascendente do NDR, que inerva a amígdala e o córtex frontal, facilitaria comportamentos defensivos aprendidos em resposta a perigo potencial ou distal, enquanto que a ativação da via periventricular, também originária no NDR e que inerva a matéria cinzenta periaquedutal dorsal (MCPD) inibiria reações de fuga e luta inatas em resposta a perigo proximal. No presente trabalho pretendeu-se explorar esta hipótese investigando o efeito da inibição do NDR, da MCPD e do núcleo amigdalóide basolateral, em ratos submetidos a dois modelos animais de ansiedade: o labirinto em T elevado (LTE) e o modelo de transição claro-escuro (MTCE). Os testes com o LTE incluíram uma variação metodológica: um dia antes do experimento, os animais foram pré-expostos por 30 minutos a um dos braços abertos do modelo. Tem sido demonstrado que a pré-exposição leva a uma diminuição do tempo gasto em atividade exploratória no braço aberto, assegurando que durante as medidas de fuga, os animais estão realmente fugindo do estímulo aversivo. No presente trabalho, a administração intra-NDR do agonista gabaérgico muscimol diminuiu as latências da esquiva inibitória (EI), efeito ansiolítico, e da fuga dos braços abertos do LTE, efeito ansiogênico. Além disso, o muscimol intra-NDR aumentou o tempo gasto no compartimento claro (TGCC) do MTCE, efeito ansiolítico. Por outro lado, a droga não apresentou efeito no modelo do labirinto em T fechado. Já a microinjeção de muscimol intra-MCPD aumentou a latência de fuga do braço aberto do LTE, efeito ansiolítico, não apresentando qualquer efeito na EI do labirinto em T elevado ou no MTCE. Finalmente, a administração de muscimol intra-amígdala basolateral prejudicou a esquiva inibitória do LTE e aumentou o TGCC do MTCE, efeitos ansiolíticos, sem alterar a fuga dos braços abertos. Em conjunto, os dados do presente trabalho corroboram alguns dos pressupostos da teoria do papel dual da 5-HT na ansiedade proposta por Deakin e Graeff (1991). É interessante salientar, ainda, a importância da pré-exposição sobre as medidas de fuga do braço aberto do LTE. / It has been proposed that distinct 5-HT pathways modulate different types of anxiety reaction to aversive stimuli. Deakin and Graeff (1991) proposed that the activation of the ascending dorsal raphe (DR)-5-HT pathways innervating the amygdala and frontal cortex would facilitate learned defensive behaviors in response to distal or potential threat, while activation of the DR-periventricular 5-HT pathways, which innervates the dorsal periaqueductal gray matter (DPAG) would inhibit innate flight or fight reactions in response to proximal threat. In an attempt to explore this hypothesis, we investigated the effects of reversible inhibition of the DRN, the DPAG and of the basolateral amygdala nucleus in rats exposed to 2 animal models of anxiety: the elevated T maze (ETM) and the light-dark model (LDM). An important methodological difference was included to the ETM tests. The animals were exposed for 30 minutes to one of the ETM open arms, 24 h before the tests. Pre-exposure seems to improve escape measurements in the ETM, assuring that during escape, animals are really escaping from the aversive threat and not simply ambling. In the present study, muscimol administered intra-DRN impaired both the avoidance, anxyolitic effect, and the escape task, anxyogenic effect, of the ETM and increased the time spent in the light compartment (TSLC) in the LDM, an anxyolitic effect. On the other hand, muscimol microinjected intra-DPAG enhanced escape latencies in the ETM, an anxyolitic effect, without altering inhibitory avoidance or LDM measurements. Finally, muscimol administered intra-basolateral amygdala impaired avoidance in the ETM and increased the TSLC in the LDM, revealing an anxyolitic effect in both tests, without altering the escape task. Taken together the obtained data seem to corroborate the dual role of 5-HT in anxiety, proposed by Deakin and Graeff (1991). At last, the importance of pre-exposure to the open arms of the LTE should be emphasized as a procedure capable of improving escape measurement.

Lopes, Marcel Adriano. Efeitos do clonazepam sobre as respostas defensivas medidas em ratos submetidos ao labirinto em T elevado. 2010. 83f. Dissertação (mestrado) - Faculdade de Medicina de Ribeirão Preto - Universidade de São Paulo - Ribeirão Preto, 2010. O labirinto em T elevado (LTE) é um modelo etológico, que evoca comportamentos defensivos correlacionados com o transtorno de ansiedade generalizada (esquiva inibitória) e o transtorno do pânico (fuga). Apesar da validação farmacológica da tarefa de esquiva inibitória deste modelo estar bem estabelecida, algumas questões em relação à tarefa de fuga não estão claras. Resultados prévios da literatura mostram que drogas clinicamente eficazes no tratamento do transtorno do pânico, como antidepressivos tricíclicos (ex: imipramina e clomipramina) e inibidores seletivos da recaptação de serotonina (ex: fluoxetina e escitalopram), aumentam a latência de fuga no LTE, sugerindo efeito do tipo panicolítico. Entretanto, em relação aos benzodiazepínicos de alta potência, também amplamente utilizados na clínica para o tratamento do transtorno do pânico, os dados em relação ao LTE permanecem desconhecidos. Sendo assim, o presente trabalho teve como objetivo investigar o efeito da administração aguda ou repetida (7 ou 14 dias) de clonazepam em ratos submetidos ao LTE. Dada a importância da substância cinzenta periaquedutal dorsal (SCPD) na gênese do transtorno do pânico, verificamos se o efeito da administração aguda de clonazepam no LTE era bloqueado pela administração prévia intra-SCPD do antagonista de receptores BZD flumazenil. Nossos resultados mostram que o tratamento agudo com clonazepam diminuiu as latências de esquiva inibitória e aumentou as latências de fuga do braço aberto, indicativo de efeito ansiolítico e panicolítico, respectivamente. Já a administração repetida deste mesmo benzodiazepínico, seja por 7 ou 14 dias, diminuiu as latências de esquiva inibitória, sem alterar as respostas de fuga. A administração intra-SCPD do antagonista de receptores benzodiazepínicos flumazenil não bloqueou o efeito da administração aguda de clonazepam sobre as respostas defensivas medidas no LTE. Os resultados do presente estudo mostram ainda que o efeito do clonazepam sobre a resposta de fuga é dependente da maneira pela qual a aquisição da resposta de esquiva inibitória é realizada, ou seja, o tratamento agudo com clonazepam foi capaz de alterar esta resposta somente quando a esquiva inibitória foi realizada com 6 tentativas. Em suma, nossos dados mostram que a administração aguda de clonazepam promove efeito panicolítico e ansiolítico no LTE. No entanto, deve ser ressaltado que o efeito panicolítico do clonazepam foi apenas observado após a introdução de uma mudança metodológica no teste do LTE. De uma maneira geral, os resultados obtidos sustentam a associação entre o comportamento de fuga e ataques de pânico. / Lopes, Marcel Adriano. Effect of clonazepam on the defensive responses measured in rats tested in the elevated T maze. 2010. 83f. Dissertação (mestrado) - Faculdade de Medicina de Ribeirão Preto - Universidade de São Paulo - Ribeirão Preto, 2010. The elevated T maze (ETM) is an ethological model that generates in rats defensive behaviors in rats which have been associated with generalized anxiety (inhibitory avoidance) and panic (escape) disorders. A wealth of evidence in the literature supports the validity of the ETM inhibitory avoidance task for detecting the effects of generalized anxiety-effective drugs. However, the effect of panic-effective drugs on escape performance has not been fully investigated yet. Previous studies showed that panic-ameliorating drugs such as tricyclic antidepressants (e.g. imipramine and clomipramine) and selective serotonin reuptake inhibitors (e.g. fluoxetine and escitalopram), increase escape latencies in the ETM, suggesting a panicolytic-like effect. The effect of high-potency benzodiazepines agonists such as clonazepam and alprazolam, also widely used for the treatment of panic disorder, remains unknown. In this study we investigated the effect of acute or repeated administration (7 or 14 days) of clonazepam in rats submitted to the ETM. Given the attributed importance of the dorsal periaqueductal gray matter (dPAG) in the genesis/regulation of panic disorder, we also investigated whether the effects caused by the acute administration of clonazepam in ETM can be blocked by prior intra-dPAG administration of the BZD receptor antagonist flumazenil. Our results showed that acute treatment with clonazepam impaired inhibitory avoidance acquisition and increased escape latencies, suggesting anxiolytic and panicolytic effects, respectively. Repeated administration of clonazepam, either for 7 or 14 days, also impaired inhibitory avoidance acquisition, but did affect escape expression. Intra-dPAG injection of flumazenil did not block the effect of clonazepam on the defensive responses measured in ETM. Our results also showed that the effect of clonazepam on the escape response is dependent on the way inhibitory avoidance acquisition is measured, i.e. the drug anti-escape effect was only observed in group of animals that had 6 but not 3 trials to acquire inhibitory avoidance. Altogether, our data show that clonazepam causes both anxiolytic and panicolytic effects on ETM, in accordance to its therapeutic profile. However, it should be emphasized that the panicolytic effect of clonazepam was only observed after the introduction of a methodological modification in the ETM test protocol. Overall, our findings support the proposed association between escape behavior and panic attacks.

Animal preferences in choice tests have frequently been used within animal welfare research to make recommendations about animal handling and husbandry. It is therefore important that these results are obtained in a way that, as far as possible, respects the behavioural capabilities of the animal. Stress has been shown to affect cognitive processes in animals and could therefore affect the decision making process. To examine the effects of stress on decision making, 16 laying hens were trained to distinguish between two different quantities of a food reward. A decision balance point was found, by increasing the cost of reaching the large reward, in lines with the theory of demand curves. Hens were then tested in a t-maze choice test with both balanced and unbalanced sets of options, with and without prior stress treatment. Choice, latency to choose, heart rate and temperatures were recorded. Hens that received stress treatment prior to their first test session were affected by this even in subsequent sessions where they did not receive stress treatment. This effect was not found in hens that first received stress treatment prior to their second test session. This shows the influence of previous experiences on animal decision making. Also, a decrease in heart rate during the decision making period was found, when making a choice between balanced options, indicating anticipation of difficulty. Additionally, this shows that physiological measurements such as heart rate could be of importance for future studies and greater understanding of underlying processes of animal decision making.

Tem sido proposto que vias serotonérgicas distintas originárias do núcleo dorsal da rafe (NDR) modulariam diferentes tipos de reações de defesa a estímulos aversivos. Deakin e Graeff (1991) propuseram que a ativação da via ascendente do NDR, que inerva a amígdala e o córtex frontal, facilitaria comportamentos defensivos aprendidos em resposta a perigo potencial ou distal, enquanto que a ativação da via periventricular, também originária no NDR e que inerva a matéria cinzenta periaquedutal dorsal (MCPD) inibiria reações de fuga e luta inatas em resposta a perigo proximal. No presente trabalho pretendeu-se explorar esta hipótese investigando o efeito da inibição do NDR, da MCPD e do núcleo amigdalóide basolateral, em ratos submetidos a dois modelos animais de ansiedade: o labirinto em T elevado (LTE) e o modelo de transição claro-escuro (MTCE). Os testes com o LTE incluíram uma variação metodológica: um dia antes do experimento, os animais foram pré-expostos por 30 minutos a um dos braços abertos do modelo. Tem sido demonstrado que a pré-exposição leva a uma diminuição do tempo gasto em atividade exploratória no braço aberto, assegurando que durante as medidas de fuga, os animais estão realmente fugindo do estímulo aversivo. No presente trabalho, a administração intra-NDR do agonista gabaérgico muscimol diminuiu as latências da esquiva inibitória (EI), efeito ansiolítico, e da fuga dos braços abertos do LTE, efeito ansiogênico. Além disso, o muscimol intra-NDR aumentou o tempo gasto no compartimento claro (TGCC) do MTCE, efeito ansiolítico. Por outro lado, a droga não apresentou efeito no modelo do labirinto em T fechado. Já a microinjeção de muscimol intra-MCPD aumentou a latência de fuga do braço aberto do LTE, efeito ansiolítico, não apresentando qualquer efeito na EI do labirinto em T elevado ou no MTCE. Finalmente, a administração de muscimol intra-amígdala basolateral prejudicou a esquiva inibitória do LTE e aumentou o TGCC do MTCE, efeitos ansiolíticos, sem alterar a fuga dos braços abertos. Em conjunto, os dados do presente trabalho corroboram alguns dos pressupostos da teoria do papel dual da 5-HT na ansiedade proposta por Deakin e Graeff (1991). É interessante salientar, ainda, a importância da pré-exposição sobre as medidas de fuga do braço aberto do LTE. / It has been proposed that distinct 5-HT pathways modulate different types of anxiety reaction to aversive stimuli. Deakin and Graeff (1991) proposed that the activation of the ascending dorsal raphe (DR)-5-HT pathways innervating the amygdala and frontal cortex would facilitate learned defensive behaviors in response to distal or potential threat, while activation of the DR-periventricular 5-HT pathways, which innervates the dorsal periaqueductal gray matter (DPAG) would inhibit innate flight or fight reactions in response to proximal threat. In an attempt to explore this hypothesis, we investigated the effects of reversible inhibition of the DRN, the DPAG and of the basolateral amygdala nucleus in rats exposed to 2 animal models of anxiety: the elevated T maze (ETM) and the light-dark model (LDM). An important methodological difference was included to the ETM tests. The animals were exposed for 30 minutes to one of the ETM open arms, 24 h before the tests. Pre-exposure seems to improve escape measurements in the ETM, assuring that during escape, animals are really escaping from the aversive threat and not simply ambling. In the present study, muscimol administered intra-DRN impaired both the avoidance, anxyolitic effect, and the escape task, anxyogenic effect, of the ETM and increased the time spent in the light compartment (TSLC) in the LDM, an anxyolitic effect. On the other hand, muscimol microinjected intra-DPAG enhanced escape latencies in the ETM, an anxyolitic effect, without altering inhibitory avoidance or LDM measurements. Finally, muscimol administered intra-basolateral amygdala impaired avoidance in the ETM and increased the TSLC in the LDM, revealing an anxyolitic effect in both tests, without altering the escape task. Taken together the obtained data seem to corroborate the dual role of 5-HT in anxiety, proposed by Deakin and Graeff (1991). At last, the importance of pre-exposure to the open arms of the LTE should be emphasized as a procedure capable of improving escape measurement.

nÃo hà / Os transtornos de ansiedade possuem alta prevalÃncia na populaÃÃo com graus de severidade variÃveis, podendo chegar à incapacitaÃÃo. Deakin & Graeff conceberam um modelo teÃrico relacionando defesa distal, amÃgdala e ansiedade generalizada, de um lado, e defesa proximal, substÃncia cinzenta periaqueductal e pÃnico, de outro. Nesta proposta, a serotonina facilita a ansiedade, porÃm inibe o pÃnico. O modelo do Labirinto em T Elevado (LTE) seria entÃo mais efetivo para distinguir estes dois padrÃes de ansiedade, ao contrario do Labirinto em Cruz Elevado (LCE), um dos modelos mais utilizados para avaliar efeitos ansiolÃticos das drogas. O modelo foi validado para ratos, havendo pouca descriÃÃo sobre o uso em camundongos. No primeiro experimento os animais (camundongos Swiss; 10 em cada grupo) foram tratados com salina (10 ml/kg; i.p.), imipramina (30mg/kg; i.p.), diazepam (1mg/kg; i.p.), paroxetina (5mg/kg; i.p.), paroxetina (10mg/kg; i.p.) e paroxetina (20mg/kg; i.p.) e avaliados no Teste do Nado ForÃado (TNF). No segundo experimento os camundongos foram separados em cinco grupos (n=10) e tratados com salina (10 ml/kg; i.p.), imipramina (30mg/kg; i.p.), diazepam (2mg/kg; i.p.), paroxetina (10mg/kg; i.p.), buspirona (10mg/kg; i.p) diariamente por uma semana (subcronicamente) e depois avaliados no TNF. No terceiro experimento camundongos foram tratados subcronicamnete com salina, imipramina (30mg/kg; IP), paroxetina (10mg/kg; IP), diazepam (2mg/kg; IP) e buspirona (10mg/kg; IP) e depois testados no LCE. No quarto experimento foram utilizados ratos Wistar que recebiam diariamente por gavagem salina, imipramina (10mg/kg), paroxetina (10mg/kg), diazepam (1mg/kg) ou buspirona (10mg/kg) por 24 dias consecutivos (volume constante 1 ml/kg de peso) e depois foram avaliados no LTE e Campo Aberto (CA). O quinto experimento foi semelhante ao quarto, sendo utilizado camundongos e um aparelho para LTE adaptado. Os resultados foram: imipramina e as trÃs dose de paroxetina apresentaram efeito antidepressivo, enquanto diazepam mostrou efeito depressivo no TNF agudo. Jà no TNF subcrÃnico apenas imipramina apresentou efeito antidepressivo. No LCE imipramina apresentou efeito ansiolÃtico enquanto paroxetina apresentou efeito ansiogÃnico. No LTE com ratos tratados cronicamente, paroxetina, diazepam e buspirona apresentaram efeito anti-ansiedade-generalizada enquanto imipramina, diazepam e paroxetina apresentaram efeito anti-pÃnico. No LTE com camundongos tratados cronicamente, imipramina, diazepam, buspirona e paroxetina apresentaram respostas anti-ansiedade-generalizada e apenas a paroxetina demonstrou efeito anti-pÃnico. Assim, imipramina, uma droga eficaz nos transtornos depressivos, ansiedade generalizada e pÃnico, foi responsÃvel por respostas semelhantes nos modelos animais (com exceÃÃo do LTE para camundongos). Diazepam, uma droga utilizada para ansiedade generalizada e com alguns efeitos no pÃnico, apresentou comportamento semelhante nos modelos, tendo efeito antipÃnico no LTE para ratos e nÃo para camundongos. Buspirona, uma droga utilizada na clinica apenas para ansiedade generalizada, nÃo apresentou efeitos antidepressivos ou antipÃnico em nenhum experimento. Paroxetina, inibidor seletivo da recaptaÃÃo da serotonina, antidepressivo, ansiolÃtico e antipÃnico, apresentou atividade ansiogÃnica no LCE (demonstrando este aparelho como inadequado para avaliar esta classe de droga) e efeito anti-ansiedade-generalizada e anti-pÃnico tanto em LTE para ratos quanto para camundongos. Conclui-se que o LTE para camundongos à um modelo adequado para detectar efeitos ansiolÃticos e anti-pÃnico das drogas, este Ãltimo em especial para drogas serotonÃrgicas / The disorders of anxiety use to have high levels of prevalence among population, with varying degrees of severity, eventually causing disability. Deakin & Graeff have conceived a theoretical model, relating distal defence, amygdala and generalized anxiety, on the one hand, and proximal defence, periaqueductal grey substance and panic, on the other. In this proposal, serotonin eases anxiety, but inhibits the panic. The format of the Elevated T Maze (ETM) would be more effective to distinguish these two patterns of anxiety, in contrast to Elevated plus maze (EPM), one of the models most frequently used to evaluate anxiolytic effects of drugs. The model was validated for rats, with little description about the use in mice. In the first experiment, the animals (Swiss mice, 10 in each group) were treated with saline (10 ml / kg, ip), imipramine (30mg/kg; ip), diazepan (1mg/kg; ip), paroxetine (5mg/kg ; Ip), paroxetine (10mg/kg; ip) and paroxetine (20mg/kg; ip) and evaluated in the Forced Swim Test (FST). In the second experiment the mice were divided into five groups (n = 10) and treated with saline (10 ml/kg, ip), imipramine (30mg/kg; ip), diazepan (2mg/kg; ip), paroxetine (10mg / kg, ip), buspirone (10mg/kg; ip), daily for one week (subchronic), and after that they were evaluated in the FST. In the third experiment mice were treated subcronic with saline, imipramine (30mg/kg; IP), paroxetine (10mg/kg; IP), diazepam (2mg/kg; IP) and buspirone (10mg/kg; IP) and then tested in EPM. In the fourth experiment, were used Wistar rats that received daily, by gavage, saline, imipramine (10mg/kg), paroxetine (10mg/kg), diazepam (1mg/kg) or buspirone (10mg/kg) for 24 consecutive days (in volume 1 ml per kg) and then were evaluated in ETM and Open Field Test (OFT). The fifth experiment was similar to the fourth, being used mice and adapted device for ETM. The results were: imipramine and three doses of paroxetine presented antidepressant effect, as diazepam showed depressive effect on acute FST. In subchronic FST, only imipramine presented antidepressant effect. In the EPM, imipramine presented anxiolytic effect as paroxetine presented anxiogenic effect. In ETM with rats chronically treated, paroxetine, diazepan and buspirone showed anti-generalized-anxiety-like effect, as imipramine, diazepan and paroxetine showed anti-panic-like effect. In ETM with mice chronically treated with imipramine, diazepan, buspirone and paroxetine presented responses anti-generalized-anxiety-like and only paroxetine showed anti-panic-like effect. Thus, imipramine, a drug effective in depressive disorders, generalized anxiety and panic, was responsible for similar responses in animal models (except for the ETM for mice). Diazepam, a drug used to generalized anxiety and with some effects in panic, presented similar behavior in the models, with antipanic-like effect in ETM for rats and not for the mice. Buspirone, a drug used in clinic only to generalized anxiety, did not present antidepressant or antipanic effects in any experiment. Paroxetine, a selective serotonin reuptake inhibitor (SSRI), antidepressant, anxiolytic and antipanic, presented anxiogenic activity in EPM (demonstrating this device as inadequate to evaluate this class of drugs) and anti-generalized-anxiety and anti-panic both in ETM for rats and for mice. We concludes that the ETM for mice is an appropriate model to detect anxiolytic and anti-panic effects of drugs, the latter especially for 5-HT drugs

Lopes, Marcel Adriano. Efeitos do clonazepam sobre as respostas defensivas medidas em ratos submetidos ao labirinto em T elevado. 2010. 83f. Dissertação (mestrado) - Faculdade de Medicina de Ribeirão Preto - Universidade de São Paulo - Ribeirão Preto, 2010. O labirinto em T elevado (LTE) é um modelo etológico, que evoca comportamentos defensivos correlacionados com o transtorno de ansiedade generalizada (esquiva inibitória) e o transtorno do pânico (fuga). Apesar da validação farmacológica da tarefa de esquiva inibitória deste modelo estar bem estabelecida, algumas questões em relação à tarefa de fuga não estão claras. Resultados prévios da literatura mostram que drogas clinicamente eficazes no tratamento do transtorno do pânico, como antidepressivos tricíclicos (ex: imipramina e clomipramina) e inibidores seletivos da recaptação de serotonina (ex: fluoxetina e escitalopram), aumentam a latência de fuga no LTE, sugerindo efeito do tipo panicolítico. Entretanto, em relação aos benzodiazepínicos de alta potência, também amplamente utilizados na clínica para o tratamento do transtorno do pânico, os dados em relação ao LTE permanecem desconhecidos. Sendo assim, o presente trabalho teve como objetivo investigar o efeito da administração aguda ou repetida (7 ou 14 dias) de clonazepam em ratos submetidos ao LTE. Dada a importância da substância cinzenta periaquedutal dorsal (SCPD) na gênese do transtorno do pânico, verificamos se o efeito da administração aguda de clonazepam no LTE era bloqueado pela administração prévia intra-SCPD do antagonista de receptores BZD flumazenil. Nossos resultados mostram que o tratamento agudo com clonazepam diminuiu as latências de esquiva inibitória e aumentou as latências de fuga do braço aberto, indicativo de efeito ansiolítico e panicolítico, respectivamente. Já a administração repetida deste mesmo benzodiazepínico, seja por 7 ou 14 dias, diminuiu as latências de esquiva inibitória, sem alterar as respostas de fuga. A administração intra-SCPD do antagonista de receptores benzodiazepínicos flumazenil não bloqueou o efeito da administração aguda de clonazepam sobre as respostas defensivas medidas no LTE. Os resultados do presente estudo mostram ainda que o efeito do clonazepam sobre a resposta de fuga é dependente da maneira pela qual a aquisição da resposta de esquiva inibitória é realizada, ou seja, o tratamento agudo com clonazepam foi capaz de alterar esta resposta somente quando a esquiva inibitória foi realizada com 6 tentativas. Em suma, nossos dados mostram que a administração aguda de clonazepam promove efeito panicolítico e ansiolítico no LTE. No entanto, deve ser ressaltado que o efeito panicolítico do clonazepam foi apenas observado após a introdução de uma mudança metodológica no teste do LTE. De uma maneira geral, os resultados obtidos sustentam a associação entre o comportamento de fuga e ataques de pânico. / Lopes, Marcel Adriano. Effect of clonazepam on the defensive responses measured in rats tested in the elevated T maze. 2010. 83f. Dissertação (mestrado) - Faculdade de Medicina de Ribeirão Preto - Universidade de São Paulo - Ribeirão Preto, 2010. The elevated T maze (ETM) is an ethological model that generates in rats defensive behaviors in rats which have been associated with generalized anxiety (inhibitory avoidance) and panic (escape) disorders. A wealth of evidence in the literature supports the validity of the ETM inhibitory avoidance task for detecting the effects of generalized anxiety-effective drugs. However, the effect of panic-effective drugs on escape performance has not been fully investigated yet. Previous studies showed that panic-ameliorating drugs such as tricyclic antidepressants (e.g. imipramine and clomipramine) and selective serotonin reuptake inhibitors (e.g. fluoxetine and escitalopram), increase escape latencies in the ETM, suggesting a panicolytic-like effect. The effect of high-potency benzodiazepines agonists such as clonazepam and alprazolam, also widely used for the treatment of panic disorder, remains unknown. In this study we investigated the effect of acute or repeated administration (7 or 14 days) of clonazepam in rats submitted to the ETM. Given the attributed importance of the dorsal periaqueductal gray matter (dPAG) in the genesis/regulation of panic disorder, we also investigated whether the effects caused by the acute administration of clonazepam in ETM can be blocked by prior intra-dPAG administration of the BZD receptor antagonist flumazenil. Our results showed that acute treatment with clonazepam impaired inhibitory avoidance acquisition and increased escape latencies, suggesting anxiolytic and panicolytic effects, respectively. Repeated administration of clonazepam, either for 7 or 14 days, also impaired inhibitory avoidance acquisition, but did affect escape expression. Intra-dPAG injection of flumazenil did not block the effect of clonazepam on the defensive responses measured in ETM. Our results also showed that the effect of clonazepam on the escape response is dependent on the way inhibitory avoidance acquisition is measured, i.e. the drug anti-escape effect was only observed in group of animals that had 6 but not 3 trials to acquire inhibitory avoidance. Altogether, our data show that clonazepam causes both anxiolytic and panicolytic effects on ETM, in accordance to its therapeutic profile. However, it should be emphasized that the panicolytic effect of clonazepam was only observed after the introduction of a methodological modification in the ETM test protocol. Overall, our findings support the proposed association between escape behavior and panic attacks.

This study investigated spatial learning in Caenorabditis elegans; the ability to associate reinforcing cues with a location. Naive, wildtype C. elegans were trained in a microfluidic t-maze in the presence of diacetyl (a volatile attractant associated with food) and subsequently tested to see if they could associate diacetyl with one arm of the t-maze. 70-80% of the subjects chemotaxed towards diacetyl during training phase, but they randomly chose left or right when diacetyl was absent (number of subjects that chose diacetyl being under 65%). From our experiments, it is unlikely that the worms are associating diacetyl with one arm of the t-maze, but appears to be using some component of the atmosphere as a cue.