Células de Schwann (CS) cultivadas a partir de nervo ciático de ratos foram enxertadas, associadas ou não ao fator de crescimento derivado de células gliais (GDNF), em ratos adultos submetidos à transecção total da medula espinhal (T11). Análises do comportamento motor do 1º ao 3º mês pós-operatório e do crescimento de fibras nervosas imunorreativas no epicentro do enxerto ao término deste período mostraram: melhora temporal dos parâmetros motores superior nos grupos que receberam CS, que foi potencializada na presença do GDNF; aumento na quantidade de axônios nos grupos que receberam CS, efeito potencializado na presença do GDNF. Em outro experimento, as reatividades astrocitária, microglial e do fator neurotrófico bFGF astrocitário foram avaliados na medula espinhal parcialmente transectada de ratos, 1 semana e 3 meses após a cirurgia. Ativação astrocitária e microglial e maior intensidade de bFGF nas substâncias branca e cinzenta foram encontradas em toda a extensão da medula, fatos relacionados ao processo local de cicatrização e fenômenos tróficos e plásticos à distância. / Schwann cells (SC) obtained from rat sciatic nerve were grafted, in the presence or not of glial derived neurotrophic factor (GDNF), in adult rats submitted to a complete spinal cord transection at T11. Behavioral analysis from the 1st to the 3rd post operative month and of the immunoreactive fiber outgrowth in the graft epicenter at the end of this period showed: temporal improvements on motor parameters that were superior in the groups that received SC, being even higher in the presence of GDNF; increased amount of axonal fibers in the groups treated with SC, being even higher in the presence of GDNF. In another experiment, astrocytic and microglial activation and astrocytic neurotrophic factor bFGF expression were analyzed after a partial spinal cord transection, 1 week and 3 months after surgery. Astrocitic and microglial activation as well as a higher intensity of astrocitic bFGF in the white and gray matters were found in the entire spinal cord, what is related to the local scar formation and distant trophic and plastic phenomena.

O glifosato é um herbicida utilizado na agricultura para o controle de plantas daninhas. No Brasil o glifosato é comercializado com vários nomes, dentre eles Roundup®; esses produtos são acrescidos de ingredientes ditos inertes que permitem a penetração do glifosato através da membrana plasmática da planta, potencializando a sua ação, além de propiciar maior estabilidade e potencial de bioacumulação. Considerando que a presença de certas substâncias químicas em formulações comerciais de glifosato pode aumentar sua toxicidade e que há carência de estudos sobre a toxicologia do desenvolvimento com essas formulações, o presente estudo investigou os efeitos da exposição perinatal de ratos ao Roundup®, a fim de avaliar o comportamento e neuroquímica cerebral materna, bem como o desenvolvimento físico e neurocomportamental da prole, desde o nascimento até a idade adulta. Para tanto, ratas prenhes receberam 50, 100 e 150 mg/kg do 15° dia de gestação até o 7° dia da lactação. Os resultados mostraram que o glifosato- Roundup® não alterou o peso materno durante os períodos de exposição ao herbicida e tampouco o consumo de água e ração das ratas, sugerindo ausência de toxicidade materna. Por outro lado, observou-se que o número de filhotes vivos de todos os grupos de ratas expostas ao herbicida foi menor do que aquele das fêmeas do grupo controle. Nas ratas expostas ao glifosato-Roundup® observou-se redução de atividade geral no campo aberto na maior dose, prejuízo no comportamento materno (CM) e no comportamento materno agressivo (CMA), bem como no hipotálamo aumento da atividade do sistema dopaminérgico e redução da atividade do sistema serotoninérgico, enquanto no hipocampo e estriato houve diminuição da atividade do sistema dopaminérgico. Esses achados sugerem que o aumento da atividade do sistema dopaminérgico hipotalâmico, por reduzir a liberação de prolactina, um hormônio essencial para a expressão do CM, prejudicou a manifestação desse comportamento. No mesmo sentido, a serotonina e dopamina por modularem os efeitos da ocitocina, um peptídeo importante no controle e na eliciação do CM e CMA, podem ter contribuído para os presentes achados. Quanto ao estudo dos parâmetros de desenvolvimento físico e neurocomportamental da prole, não foram modificados pela administração do herbicida. No presente trabalho foi avaliada também a atividade geral da prole na infância e na idade adulta, mostrando que as alterações foram pontuais tanto na prole masculina como na feminina, não caracterizando alterações motoras ou emocionais. Quanto ao comportamento de brincar de luta, a administração do glifosato-Roundup® reduziu apenas o pinning tanto de ratos machos como de fêmeas, sem alterar outros parâmetros deste comportamento. A duração e frequência do pinning são consideradas como parâmetros que expressam o início do comportamento social do rato. Desta forma, nestes animais não foram estabelecidos os padrões de comportamento social durante o desenvolvimento, porém aqueles relacionados com o comportamento de luta e sexual foram preservados. Na idade adulta, não foram observadas alterações na interação social de ratos e ratas expostos ao herbicida, ou seja, não promoveu efeitos em longo prazo no comportamento social. Quanto aos achados de neuroquímica cerebral da prole adulta exposta ao herbicida, foram encontradas alterações nos diferentes sistemas de neurotransmissão, contudo a ausência de alterações relevantes na atividade geral e na interação social das proles masculina e feminina adultas, não foi possível especular sobre o envolvimento desses sistemas de neurotransmissão com esses comportamentos. Esses achados em conjunto, mostraram que a exposição perinatal a diferentes doses de glifosato- Roundup® promoveu prejuízo no comportamento materno, os quais foram associados aos sistemas de neurotrnasmissão cerebrais, e na prole, embora não tenham sido observados efeitos no desenvolvimento físico e neurocomportamental, ocorreram alterações comportamentais na infância que se estenderam até a idade adulta. / Glyphosate is a herbicide used in agriculture to control weeds. In Brazil glyphosate is marketed under several names, including Roundup ®. Ingredients considered inert are added to these products in order to allow the penetration of glyphosate across the plasma membrane of the plant, increasing its action, as well as providing greater stability and bioaccumulation potential. Considering that the presence of certain chemicals in commercial formulations of glyphosate may increase its toxicity and that there are insufficient data on the toxicology of development using these formulations, the present study investigated the effects of perinatal exposure of rats to Roundup®, to assess behavior and maternal brain neurochemistry, as well as physical and neurobehavioral development of the offspring from birth to adulthood. To this end, pregnant rats received 50, 100 and 150 mg/kg from the 15th gestation day to the 7th lactation day. The results showed that glyphosate-Roundup® did not alter maternal weight during neither periods of exposure to the herbicide nor the consumption of water and feed the rats, suggesting the absence of maternal toxicity. On the other hand, it was observed that the number of living offspring of all groups of rats exposed to herbicide was smaller than that of the female control group. In rats exposed to glyphosate-Roundup® it was possible to observe a reduction in general activity in the open field at the highest dose, impairment in maternal behavior (CM) and maternal aggressive behavior (CMA), as well as in the hypothalamus there was an increase in the dopaminergic system activity and a reduction of the serotonergic system activity, whereas the hippocampus and striatum showed decreased activity of the dopaminergic system. These findings suggest that the increased activity of the hypothalamic dopaminergic system by reducing the release of prolactin, an essential hormone for the expression of CM, damaged the manifestation of this behavior. Similarly, serotonin and dopamine, which modulate the effects of oxytocin, an important peptide in the control and elicitation of the CM and CMA, may have contributed to the present findings. Regarding the study of the parameters of physical and neurobehavioral offsprings development, no modification was caused by the administration of the herbicide. The present study also evaluated the overall activity of the offspring in childhood and adulthood, showing that the changes were punctual in both male and female offspring in not characterizing motor or emotional changes. Regarding the behavior of play fighting, the presence of glyphosate-Roundup® reduced only the pinning from both male and female rats, without changing other parameters of this behavior. The duration and frequency of pinning are considered parameters which express the beginning of the social behavior of the rats. Thus, during the development of these animals, the patterns of social behavior have not been established, but those related to fighting and sex behavior were preserved. In adulthood, no changes were observed in the social interaction of male and female rats exposed to the herbicide, i. e., it did not promote long-term effects on social behavior. In the present study, considering the findings of brain neurochemistry of adult offspring exposed to the herbicide, it was possible to observe in the hypothalamus an increase in dopamine turnover in both male and female adult offspring and an increased turnover of serotonin only in male offspring. In the hippocampus, the most relevant findings were increased turnover of serotonin in males and in females the opposite effect. In the striatum an increased turnover of serotonin was found in both males and females, and dopamine only in females. Taking these data together and considering the absence of material changes in general activity measured in the open field and social interaction of male and female adult offspring, it is not possible to speculate about the relation of these neurotransmitter systems with these behaviors.

A ivermectina é uma lactona macrocíclica utilizada para o tratamento de parasitoses na espécie humana e amplamente empregada em medicina veterinária como endectocida. Em mamíferos, diversas evidências indicam que as lactonas macrocíclicas interagem com canais de cloro mediados pelo ácido gama-aminobutírico (GABA). Sabe-se que o sistema GABAérgico está envolvido com a manifestação do comportamento sexual. Assim, no presente trabalho foram estudados, em ratos, os efeitos da ivermectina na coordenação motora, na motivação e no desempenho sexual de machos, bem como no comportamento sexual de fêmeas; além disso, avaliaram-se os níveis séricos de testosterona e as concentrações de diferentes neurotransmissores e seus metabólitos no hipotálamo e estriado, áreas do sistema nervoso central relacionadas, respectivamente, com o comportamento sexual e atividade motora. No presente estudo foram usadas as doses de 0,2 e 1,0 mg/kg, por via intraperitoneal, de uma formulação comercial de ivermectina; estas doses foram escolhidas considerando que a primeira é a dose usualmente empregada terapeuticamente e a segunda baseada em achados prévios que mostraram interferência no comportamento sexual de ratos machos. Os resultados mostraram que a administração de ivermectina em ratos: promoveu prejuízo na coordenação motora de machos; não interferiu na motivação sexual e na ereção peniana de machos; prejudicou o comportamento sexual de ratas; não alterou o peso relativo dos testículos, epidídimos, próstata e vesícula seminal, porém aumentou o peso relativo do fígado; não alterou o índice gonadossomático de machos, bem como não foram observadas alterações no estudo histopatológico dos diferentes órgãos; reduziu os níveis séricos de testosterona de machos; diminuiu os níveis de GABA hipotalâmico e estriatal, bem como reduziu a atividade dos sistemas serotoninérgico e dopaminérgico hipotalâmicos e, no estriado, reduziu a atividade do sistema dopaminérgico. Os resultados foram discutidos considerando a interferência da ivermectina nos sistemas de neurotransmissores central. / Ivermectin is a macrocyclic lactone used for the treatment of parasitic infections in humans and widely used in veterinary medicine as endectocide. In mammals, a number of evidence indicate that interact with the macrocyclic lactones chloride channels mediated by gamma-aminobutyric acid (GABA). It is known that the GABAergic system is involved in the manifestation of sexual behavior. Thus, in the present work were studied in rats, the effects of ivermectin on motor coordination, motivation and sexual performance of males and females in sexual behavior; furthermore, evaluated the serum levels of testosterone and different concentrations of neurotransmitters and their metabolites in the hypothalamus and striatum areas of the central nervous system related, respectively, sexual behavior and motor activity. In the present study doses of 0.2 and 1.0 mg / kg were used, intraperitoneally, a commercial formulation of ivermectin; these doses were chosen considering that the first dose is usually employed therapeutically and the second based on previous findings have shown that interference with sexual behavior of male rats. The results showed that administration of ivermectin in rats: impairment in motor coordination promoted male; did not interfere on the sexual motivation and the penile erection in male; impaired sexual behavior of female rats; did not alter the relative weight of the testes, epididymis, prostate and seminal vesicles, but increased the relative liver weight; did not alter the gonadosomatic index (GSI) of males and no changes were observed in the histopathological study of different organs; reduced serum testosterone levels of male; decreased levels of hypothalamic and striatal GABA and reduced the activity of the serotonergic and dopaminergic systems hypothalamic and the, reduced the activity of the dopaminergic system striatal. The results were discussed considering the interference of ivermectin in central neurotransmission systems.

O núcleo acumbens (Acb) é envolvido em comportamentos adaptativos e emocionais. A maioria dos neurônios do Acb são neurônios de projeção (MSNs) que contém a fosfoproteína DARPP-32 e são modulados por distintos tipos de interneurônios. Há pouca informação sobre os receptores de glutamato (Glu) expressos no Acb. Este estudo investiga, através de técnicas de imunohistoquímica, a distribuição e co-localização de marcadores do sistema dopaminérgico, dos receptores de Glu do tipo AMPA (GluR1-GluR4) e NMDAR1 e marcadores de interneurônios. Nossos resultados mostram que o Acb possui uma neuroquímica semelhante ao estriado dorsal. Porém, detectamos uma distribuição distinta de alguns dos marcadores no Acb. Os estudos de co-localização revelam que quase todos os neurônios no Acb expressam GluR2/3 ou GluR2. Em contraste, GluR1 e GluR4 são fracamente expressas e co-localizam com parvalbumina. Esses resultados indicam que GluR2 e GluR2/3 são expressas em MSNs DARPP-32+ e na maioria dos interneurônios do Acb enquanto GluR1 e GluR4 são exclusivamente expressas em interneurônios. / The nucleus accumbens is involved in adaptive and emotional behaviors. The majority of neurons in the Acb are projection neurons that express the phosphoprotein DARPP-32 and are modulated by distinct types of interneurons. There is little information about the glutamate receptors expressed in the Acb. This study investigates by immunohistochemical methods the distribution and co-localization of markers of the dopaminergic system, AMPA (GluR1-4) and NMDAR1 type Glu receptor subunits and specific markers of interneurons. Our results show that the neurochemistry of the Acb is similar to that of the dorsal striatum. However, we detected a distinct distribution of some markers in the Acb. Our co-localization studies reveal that almost all neurons of the Acb express GluR2/3 or GluR2. In contrast, GluR1 and GluR4 are weakly expressed and are co-localized with parvalbumin. These results indicate that GluR2 and GluR2/3 are expressed by MSNs DARPP-32+ and by the majority of interneurons of the Acb, whereas GluR1 and GluR4 are exclusively expressed by interneurons.

O estresse destaca-se como um importante fator de risco para o desenvolvimento de diferentes doenças. Reconhecido atualmente como uma epidemia global pela Organização Mundial da Saúde e afetando mais de 90% da população mundial, o estresse apresenta grande associação, em particular, com os transtornos mentais. Dentre esses, a depressão se sobressai afetando sozinha cerca de 350 milhões de pessoas em todo mundo. Poucas experiências talvez sejam tão comuns entre os organismos quanto a exposição a eventos estressantes Alguns tipos de estresses emocionais, como a tristeza, e outros no qual o organismo é afetado por períodos prolongados, tem demonstrado serem capazes de promover disfunções imunes e distúrbios comportamentais que podem ser compreendidos através do campo interdisciplinar de estudo da neuroimunomodulação, uma vez que podem desencadear respostas específicas no eixo hipotálamo-hipófise-adrenal, que, por conseguinte, podem modular diferentes efeitos fisiológicos decorrentes da exposição ao estresse. Dentre esses efeitos, é possível destacar a capacidade de resiliência e / ou resistência ao estresse, que podem conferir uma capacidade diferenciada de recuperação, como também aumento na susceptibilidade a doenças. Considerando os aspectos distintos do estresse e os diferentes estados patológicos a ele associados, esse trabalho teve como objetivo avaliar os efeitos comportamentais, neuroquímicos e imunes do estresse de contenção de duas horas em camundongos, selecionados para um perfil distinto de reatividade ao estresse inescapável pelo teste de suspensão da cauda. Para isso, camundongos machos de alta e baixa imobilidade foram previamente selecionados e submetidos diferentes testes pré e pós exposição ao estresse de contenção de duas horas: (i) análise dos comportamentos tipo-depressivos e ansiosos, (ii) análise dos neurotransmissores no córtex pré-frontal, hipotálamo e mesencéfalo e, (iii) análise de citocinas pró- inflamatórias no córtex pré-frontal e hipotálamo. Nossos resultados mostraram que animais de alta e baixa imobilidade apresentam comportamentos diferentes antes e após a exposição ao estresse, além de apresentar um comportamento de grooming distinto após o estresse de contenção. A exposição ao estresse também promoveu alterações entre as concentrações serotoninérgicas, dopaminérgicas e noradrenérgicas entre animais de alta e baixa imobilidade, tanto no córtex pré-frontal, hipotálamo e mesencéfalo. Além disso, o perfil imune também se revelou alterado entre esses animais, principalmente em TNF- no hipotálamo, revelando uma ativação dos sistemas relacionados ao estresse. Considerando o conjunto dos dados apresentados, nossos resultados sugerem uma ativação diferenciada do eixo hipotálamo-hipófise-adrenal entre camundongos de alta e baixa imobilidade e, ainda, que os animais de baixa imobilidade apresentam um perfil de resiliência ao estresse contenção, tendo animais de alta imobilidade o perfil oposto / Stress stands out as an important risk factor for the development of different diseases. Currently recognized as a global epidemic by the World Health Organization and affecting more than 90% of the world population, stress can be strongly associated with mental disorders. Among these, depression can be highlight affecting alone about 350 million people worldwide. Few types of emotional stresses, such as sadness, and others which the organism is affected for long periods, have demonstrated to be capable of promoting immune dysfunctions and behavioral disorders. These changes can be understood through the interdisciplinary field of study of the neuroimmunomodulation, since it can trigger specific responses in the hypothalamic-pituitary-adrenal axis, which, therefore, could modulate different physiological effects resulting from the exposure to stress. Among these effects, it is possible to highlight the capacity of resilience and / or resistance to stress, which can confer a differentiated capacity of recovery, as well as increase in the susceptibility to diseases. Considering the distinct aspects of stress and the different pathological conditions associated, this study aimed to evaluate the behavioral, neurochemical and immune effects of two-hour of restraint stress in mice selected for a different profile of stress reactivity to the inescapable stress of the tail suspension test. For this, male mice of high and low immobility were previously selected and submitted to different tests before and after exposure to a two-hour restraint stress protocol for the analysis of: (i) the depressive and anxiety-like behaviors; (ii) concentrations of neurotransmitters in the prefrontal cortex, hypothalamus and midbrain; (iii) expression of proinflammatory cytokines in the prefrontal cortex and hypothalamus. Our results showed that animals of high and low immobility presented different behavioral profiles before and after exposure to stress and presented a distinct grooming behavior after the restraint stress. Exposure to stress also promoted changes between the serotonergic, dopaminergic and noradrenergic concentrations between animals of high and low immobility in the prefrontal cortex, hypothalamus and midbrain. In addition, the immune profile revealed to be altered among these animals, especially in TNF- in the hypothalamus, showing an activation of stress-related systems. Considering the set of data presented, our results suggest a differentiated activation of the hypothalamic-pituitary-adrenal axis between mice of high and low immobility and, also, that the animals of low immobility present a resilience profile to the restraint stress, having high immobility animals the opposite profile

帕金森病（PD）是一種常見的神經退行性疾病，其特徵性的癥狀是運動功能減弱，常伴有認知障礙如工作記憶缺陷。大多數癥狀源於中腦多巴胺神經元的進行性缺失。目前的治療常隨時間進展誘發嚴重的副反應，促使我們進一步研究PD的病理生理學機制。一般認為基底神經節直接和間接通路不平衡的活動導致PD的運動缺陷，但目前關於基底神經節環路突觸特性改變的研究還很少。對PD認知障礙機制的研究則更為少見。 / 本研究中，我們首先關注在對基底神經節提供主要輸入的皮質紋狀體通路。應用全細胞膜片鉗技術結合皮質刺激，并利用在D2受體表達神經元表達綠色螢光蛋白的轉基因小鼠，我們發現PD狀態下，在間接通路表達D2受體的中型多棘神經元（D2 MSN）上記到的皮質紋狀體通路AMPA受體介導電流的成對脉沖比值（PPR）以及NMDA受體介導電流的PPR均降低。此外皮質至D2 MSN突觸間隙的谷氨酸水平也增加而不伴有谷氨酸轉運體的功能異常。這些結果證明PD狀態下皮質至間接通路D2 MSN的谷氨酸釋放選擇性增加。結合基底神經節的功能環路考慮，至間接通路紋狀體投射神經元的皮質谷氨酸釋放增加可能參與了PD的運動癥狀。 / 我們接下來研究了皮質-D2 MSN通路的下游環節即紋狀體至蒼白球通路傳遞的改變。在蒼白球（GP）神經元上應用全細胞膜片鉗記錄結合紋狀體刺激，我們發現在6-羥多巴損毀之後，紋狀體蒼白球通路的PPR降低，GP神經元記到的紋狀體刺激誘發的抑制性突觸后電流（eIPSC）的變異係數降低,以及GP神經元記到的微型IPSC的頻率增加，這些結果證明紋狀體至蒼白球的GABA釋放增加。突觸前III型代謝型谷氨酸受體介導的對紋狀體蒼白球傳遞的抑制作用消失導致了紋狀體蒼白球通路GABA釋放的增加。這一增加，通過影響間接通路的下游環節，也可能參與了PD的運動癥狀。 / 為探討認知障礙的機制，我們研究了參與工作記憶功能的邊緣前皮質至伏核（NAc）的投射。應用與第一部份相似的研究方法，我們發現多巴胺受體對邊緣前皮質NAc通路的傳遞存在高度精確和補償性的調節。在邊緣前皮質-NAc D1 MSN通路，D1和D2受體突觸前分別介導對該傳遞的抑制性和易化性調節。然而，在D2 MSN相關的邊緣前皮質-NAc通路，上述作用發生了反轉。在耗竭NAc多巴胺之後，D2 MSN上誘發到的興奮性突觸后電流增加，提示邊緣前皮質-NAc D2 MSN傳遞增加。此外，在多巴胺損毀的情況下，激活D1和D2受體不再調節邊緣前皮質NAc通路的傳遞。結合邊緣環路考慮，邊緣前皮質至D2 MSN的谷氨酸釋放增加可能參與了PD的認知障礙。 / 綜上所述，PD狀態下，繼多巴胺缺失之後，多條通路發生可塑性改變，這些改變可能參與PD的運動和認知癥狀。 / Parkinson’s disease (PD) is a common neurodegenerative disease with characteristic hypokinetic motor symptoms and cognitive impairments like working memory deficits. Most of the symptoms are derived from progressive loss of dopaminergic neurons in the midbrain. Current therapies often induce severe side effects with time, which promotes us to further investigate the pathophysiological mechanism of PD. It is generally thought that the imbalanced activity between direct and indirect pathways of the basal ganglia underlies the motor deficits in PD, but little is studied about the changes in synaptic properties of the sub-circuits. Even less is known about the mechanism responsible for the cognitive dysfunctions in PD. / In our study, we first focused on the corticostriatal pathway that provides a major input to the basal ganglia. Employing whole-cell patch-clamp recordings with cortical stimulation as well as by taking advantage of transgenic mice with green fluorescent protein co-expressed in the D2 receptor-expressing neurons, we found a selective increase in cortical glutamate release onto indirect-pathway D2 receptor-expressing medium-sized spiny neurons (D2 MSNs), as indicated by reduced corticostriatal AMPA paired-pulse ratios (PPRs) and NMDA PPRs in D2 MSNs as well as increased glutamate level in cortex-D2 MSN synaptic cleft without malfunction in glutamate transporters in parkinsonian states. Considering from the functional organization of the basal ganglia circuits, the increased corticostriatal glutamate release onto indirect-pathway striatal projection neurons may contribute to the motor symptoms of PD. / We next studied whether the striatopallidal transmission, downstream to the cortex-D2 MSNs pathway, is also altered in parkinsonian states. Combining whole-cell patch-clamp recordings in globus pallidus (GP) neurons with striatal stimulation, we demonstrated that the striatopallidal GABA release was increased following 6-hydroxydopamine lesion, as indicated by decreased striatopallidal PPRs, reduced coefficient of variation of striatally evoked inhibitory postsynaptic currents (eIPSCs) and elevated frequency of miniature IPSCs in GP neurons. The loss of tonic presynaptic group III metabotropic glutamate receptors-mediated inhibition on striatopallidal transmission accounted for the increased striatopallidal GABA release. The increase in the striatopallidal GABA release, through affecting the downstream of the indirect pathway, would also contribute to the motor symptoms in PD. / To investigate the underlying mechanism of cognitive deficits, we targeted the prelimbic cortex-nucleus accumbens (NAc) projection that is critical for working memory function. Using similar approaches as the first part, we observed highly precise and complementary modulations by dopamine receptors, with D1 and D2 receptors presynaptically mediating the inhibition and facilitation of the prelimbic cortex-NAc D1 MSN transmission, respectively, and reversed effects in D2 MSN-associated pathway. Following dopamine depletion in NAc, an enhanced prelimbic cortex-NAc D2 MSN transmission was indicated by selectively increased excitatory postsynaptic current evoked in D2 MSNs. Moreover, in the dopamine-depleted state, activating D1 and D2 receptors failed to modulate the prelimbic cortex-NAc transmission. Considering from the information flow in the limbic loop, the increased prelimbic cortical glutamate release onto D2 MSNs may contribute to the cognitive impairments in PD. / In conclusion, in the parkinsonian states, multiple pathways undergo plasticity changes subsequent to dopamine depletion, which may underlie the motor and cognitive symptoms in PD. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Detailed summary in vernacular field only. / Cui, Qiaoling. / Thesis (Ph.D.)--Chinese University of Hong Kong, 2012. / Includes bibliographical references (leaves 160-191). / Electronic reproduction. Hong Kong : Chinese University of Hong Kong, [2012] System requirements: Adobe Acrobat Reader. Available via World Wide Web. / Abstract also in Chinese. / Chapter Chapter 1 --- General introduction --- p.1 / Chapter 1.1 --- Parkinson’s disease --- p.1 / Chapter 1.1.1 --- Symptoms --- p.1 / Chapter 1.1.2 --- Etiology --- p.1 / Chapter 1.1.3 --- Pathology and pathophysiology --- p.2 / Chapter 1.1.4 --- Therapy --- p.5 / Chapter 1.1.4.1 --- L-DOPA and dopamine receptor agonists treatments --- p.5 / Chapter 1.1.4.2 --- Deep brain stimulation (DBS) and lesional surgery treatments --- p.6 / Chapter 1.1.4.3 --- Neural transplantation --- p.7 / Chapter 1.1.4.4 --- Treatment of nonmotor symptoms --- p.8 / Chapter 1.2 --- Basal ganglia --- p.8 / Chapter 1.2.1 --- Components of basal ganglia --- p.8 / Chapter 1.2.2 --- Pathways in basal ganglia --- p.8 / Chapter 1.2.2.1 --- Anatomical organization of the basal ganglia pathways --- p.8 / Chapter 1.2.2.2 --- Functional consequences of the basal ganglia pathways --- p.10 / Chapter 1.3 --- Striatum --- p.11 / Chapter 1.3.1 --- Anatomy of the striatum --- p.11 / Chapter 1.3.1.1 --- Cellular heterogeneity in the striatum --- p.12 / Chapter 1.3.1.1.1 --- MSNs --- p.12 / Chapter 1.3.1.1.1.1 --- Subpopulations --- p.13 / Chapter 1.3.1.1.1.2 --- Morphology --- p.13 / Chapter 1.3.1.1.1.3 --- Electrophysiological properties --- p.14 / Chapter 1.3.1.1.2 --- Cholinergic interneurons --- p.15 / Chapter 1.3.1.1.3 --- GABAergic interneurons --- p.17 / Chapter 1.3.1.2 --- Innervation of the striatum --- p.18 / Chapter 1.3.1.3 --- Output of the striatum --- p.21 / Chapter 1.3.2 --- Function of the striatum --- p.21 / Chapter 1.3.2.1 --- Function of the associative striatum --- p.22 / Chapter 1.3.2.2 --- Function of the sensorimotor striatum --- p.22 / Chapter 1.3.3 --- The corticostriatal system --- p.23 / Chapter 1.3.3.1 --- Anatomy of the corticostriatal system --- p.24 / Chapter 1.3.3.2 --- Physiology of the corticostriatal system --- p.25 / Chapter 1.3.3.3 --- Function of the corticostriatal system --- p.26 / Chapter 1.3.4 --- Striatum, corticostriatal system and PD --- p.26 / Chapter 1.4 --- GPe --- p.29 / Chapter 1.4.1 --- Anatomy of GPe --- p.29 / Chapter 1.4.1.1 --- Cellular heterogeneity in GPe --- p.29 / Chapter 1.4.1.2 --- Innervation of GPe --- p.31 / Chapter 1.4.1.3 --- Output of GPe --- p.33 / Chapter 1.4.2 --- Neurotransmission in GPe --- p.34 / Chapter 1.4.2.1 --- GABAA receptors in GPe --- p.34 / Chapter 1.4.2.2 --- GABAB receptors in GPe --- p.35 / Chapter 1.4.2.3 --- Evoked responses in GPe from direct striatal and pallidal stimulations --- p.37 / Chapter 1.4.3 --- GPe, striatopallidal system and PD --- p.38 / Chapter 1.5 --- NAc --- p.40 / Chapter 1.5.1 --- Anatomy of NAc --- p.40 / Chapter 1.5.1.1 --- Subregions --- p.40 / Chapter 1.5.1.2 --- Cell heterogeneity in NAc --- p.42 / Chapter 1.5.1.2.1 --- MSNs --- p.42 / Chapter 1.5.1.2.2 --- Interneurons --- p.42 / Chapter 1.5.1.3 --- Inervation of NAc --- p.43 / Chapter 1.5.1.4 --- Output of NAc --- p.43 / Chapter 1.5.2 --- Function of NAc --- p.43 / Chapter 1.5.3 --- The prefrontal cortex (PFC)-NAc system --- p.43 / Chapter 1.5.4 --- NAc, PFC-NAc system and PD --- p.44 / Chapter 1.6 --- Objectives --- p.45 / Chapter Chapter 2 --- General methods --- p.51 / Chapter 2.1 --- Electrophysiological experiments --- p.51 / Chapter 2.1.1 --- Slice preparation --- p.51 / Chapter 2.1.2 --- Whole-cell patch-clamp recordings --- p.52 / Chapter 2.1.3 --- Uncaging experiment --- p.54 / Chapter 2.1.4 --- Data analysis and statistics --- p.54 / Chapter 2.2 --- Dopamine depletion --- p.55 / Chapter 2.2.1 --- 6-hydroxydopamine (6-OHDA) injection into medial forebrain bundle (MFB) --- p.55 / Chapter 2.2.2 --- 6-OHDA injection into NAc --- p.56 / Chapter 2.2.3 --- Reserpine treatment --- p.56 / Chapter 2.3 --- Limb-use asymmetry test (cylinder test) --- p.57 / Chapter 2.4 --- Tyrosine hydroxylase (TH) immunohistochemistry and analysis --- p.57 / Chapter 2.4.1 --- TH immunohistochemistry of SNc and striatal slices --- p.57 / Chapter 2.4.2 --- TH immunohistochemistry and analysis of NAc slices --- p.58 / Chapter 2.5 --- Tracing study --- p.59 / Chapter 2.6 --- Genotyping and quantitative polymerase chain reaction (qPCR) --- p.60 / Chapter Chapter 3 --- Alteration of corticostriatal glutamatergic transmission onto D2 MSNs in PD models --- p.62 / Chapter 3.1 --- Summary --- p.62 / Chapter 3.2 --- Introduction --- p.63 / Chapter 3.3 --- Materials --- p.65 / Chapter 3.3.1 --- Animals --- p.65 / Chapter 3.3.2 --- Chemicals --- p.66 / Chapter 3.4 --- Results --- p.66 / Chapter 3.4.1 --- Comparison of corticostriatal paired-pulse ratios (PPRs) between hemizygotes and homozygotes of D2-EGFP BAC transgenic mice --- p.66 / Chapter 3.4.2 --- Corticostriatal AMPA PPR was specifically decreased in D2 MSNs following dopamine depletion --- p.67 / Chapter 3.4.2.1 --- Corticostriatal AMPA PPR was specifically decreased in D2 MSNs following reserpine treatment --- p.67 / Chapter 3.4.2.2 --- Corticostriatal AMPA PPR was specifically decreased in D2 MSNs following 6-OHDA lesion --- p.68 / Chapter 3.4.3 --- Increased glutamate release underlying reduction of corticostriatal PPR in D2 MSNs in parkinsonian states --- p.69 / Chapter 3.4.3.1 --- Effect of γ-DGG on the corticostriatal eEPSCs of D2 MSNs --- p.70 / Chapter 3.4.3.2 --- Effect of γ-DGG on the corticostriatal eEPSCs of D2 MSNs in the presence of CTZ --- p.70 / Chapter 3.4.3.3 --- Decay kinetics of eEPSCs of D2 MSNs in the presence of CTZ or PEPA were not consistently altered following dopamine depletion --- p.71 / Chapter 3.4.3.4 --- Corticostriatal NMDA PPR was decreased in D2 MSNs following dopamine depletion --- p.72 / Chapter 3.4.4 --- AMPA receptor occupancy was increased in D2 MSNs following dopamine depletion --- p.73 / Chapter 3.4.5 --- Increased postsynaptic AMPA receptor desensitization contributing to the reduction of corticostriatal PPR in D2 MSNs of parkinsonian states --- p.74 / Chapter 3.4.5.1 --- Effect of CTZ on the corticostriatal AMPA PPR of D2 MSNs --- p.74 / Chapter 3.4.5.2 --- Effect of PEPA on the corticostriatal AMPA PPR of D2 MSNs --- p.75 / Chapter 3.4.6 --- Loss of dopamine D2 receptor activation did not contribute to the increased corticostriatal glutamate release onto D2 MSNs in the parkinsonian states --- p.75 / Chapter 3.4.7 --- Postsynaptic Ca2+ involved in the modification of the corticostriatal transmission in D2 MSNs of parkinsonian state --- p.77 / Chapter 3.5 --- Discussion --- p.78 / Chapter 3.5.1 --- Corticostriatal glutamate release onto D2 MSNs was increased in the parkinsonian states --- p.78 / Chapter 3.5.2 --- AMPA receptor occupancy was increased in D2 MSNs following dopamine depletion --- p.80 / Chapter 3.5.3 --- Postsynaptic AMPA receptor desensitization was increased in D2 MSNs following dopamine depletion --- p.81 / Chapter 3.5.4 --- Loss of dopamine D2 receptor activation did not contribute to the increased corticostriatal glutamate release onto D2 MSNs in the parkinsonian states --- p.81 / Chapter 3.5.5 --- Postsynaptic Ca2+ involved in the modification of the corticostriatal transmission in D2 MSNs of parkinsonian state --- p.82 / Chapter 3.5.6 --- The increased corticostriatal glutamate release onto D2 MSNs and PD --- p.83 / Chapter Chapter 4 --- Alteration of striatopallidal GABAergic transmission in 6-OHDA lesioned PD model --- p.98 / Chapter 4.1 --- Summary --- p.98 / Chapter 4.2 --- Introduction --- p.99 / Chapter 4.3 --- Materials --- p.101 / Chapter 4.3.1 --- Animals --- p.101 / Chapter 4.3.2 --- Chemicals --- p.101 / Chapter 4.4 --- Results --- p.101 / Chapter 4.4.1 --- Striatopallidal paired-pulse ratio (PPR) was decreased following 6-OHDA lesion --- p.101 / Chapter 4.4.1.1 --- Striatopallidal PPR was unchanged following reserpine treatment --- p.102 / Chapter 4.4.1.2 --- Striatopallidal PPR was decreased following 6-OHDA lesion --- p.102 / Chapter 4.4.2 --- Increased striatopallidal GABA release underlying the reduction of striatopallidal PPR following 6-OHDA lesion --- p.103 / Chapter 4.4.2.1 --- CV of eIPSC1 in GP neurons was reduced following 6-OHDA lesion --- p.103 / Chapter 4.4.2.2 --- mIPSCs frequency was increased in GP neurons following 6-OHDA lesion --- p.104 / Chapter 4.4.3 --- Mechanism for the increased striatopallidal GABA release following 6-OHDA lesion --- p.105 / Chapter 4.4.3.1 --- Loss of dopamine D2 receptor activation did not contribute to the increased striatopallidal GABA release following 6-OHDA lesion --- p.105 / Chapter 4.4.3.2 --- GABAB receptor modulation did not contribute to the increased striatopallidal GABA release following 6-OHDA lesion --- p.106 / Chapter 4.4.3.3 --- Loss of presynaptic tonic group III mGluR inhibition accounted for the increased striatopallidal GABA release following 6-OHDA lesion --- p.107 / Chapter 4.5 --- Discussion --- p.109 / Chapter 4.5.1 --- Striatopallidal GABA release was increased in the parkinsonian state --- p.109 / Chapter 4.5.2 --- Mechanism underlying the increased striatopallidal GABA release in the parkinsonian state --- p.111 / Chapter 4.5.2.1 --- Loss of dopamine D2 receptor activation did not contribute to the increased striatopallidal GABA release following 6-OHDA lesion --- p.111 / Chapter 4.5.2.2 --- GABAB receptor modulation did not contribute to the increased striatopallidal GABA release following 6-OHDA lesion --- p.112 / Chapter 4.5.2.3 --- Loss of presynaptic tonic group III mGluR inhibition accounted for the increased striatopallidal GABA release following 6-OHDA lesion --- p.113 / Chapter 4.5.3 --- The increased striatopallidal GABA release and PD --- p.114 / Chapter 4.5.4 --- The striatopallidal group III mGluR system and PD --- p.116 / Chapter Chapter 5 --- Role of D1 and D2 receptors in prelimbic cortex-nucleus acumbens transmission in normal and parkinsonian states --- p.128 / Chapter 5.1 --- Summary --- p.128 / Chapter 5.2 --- Introduction --- p.129 / Chapter 5.3 --- Materials --- p.131 / Chapter 5.3.1 --- Animals --- p.131 / Chapter 5.3.2 --- Chemicals --- p.131 / Chapter 5.4 --- Results --- p.132 / Chapter 5.4.1 --- Prelimbic cortex innervated both D1 MSNs and D2 MSNs in core subregion of NAc- --- p.132 / Chapter 5.4.2 --- D1 and D2 receptors presynaptically modulated the D1 MSN-associated prelimbic cortex-NAc transmission in opposite manner --- p.133 / Chapter 5.4.3 --- D1 and D2 receptors presynaptically modulated the D2 MSN-associated prelimbic cortex-NAc transmission in a reverse manner --- p.134 / Chapter 5.4.4 --- Effects of D1 and D2 receptor antagonists on the prelimbic cortex-Nac transmission --- p.135 / Chapter 5.4.5 --- Basal synaptic transmission was enhanced in D2 MSN-associated prelimbic cortex-NAc pathway following NAc dopamine depletion --- p.136 / Chapter 5.4.6 --- D1 and D2 receptor modulation of the prelimbic cortex-NAc transmission disappeared following dopamine depletion --- p.137 / Chapter 5.5 --- Discussion --- p.138 / Chapter 5.5.1 --- Prelimbic cortex innervated both D1 MSNs and D2 MSNs in core subregion of NAc- --- p.138 / Chapter 5.5.2 --- Prelimbic cortex-NAc projections were presynaptically modulated by D1 and D2 receptors in a highly precise and complementary pattern --- p.138 / Chapter 5.5.3 --- Glutamatergic transmission was selectively enhanced in D2 MSN-associated prelimbic cortex-NAc pathway following NAc dopamine depletion --- p.140 / Chapter 5.5.4 --- D1 and D2 receptor modulation of the prelimbic cortex-NAc transmission was lost following dopamine depletion --- p.142 / Chapter Chapter 6 --- General discussion --- p.154 / Chapter 6.1 --- Enhanced corticostriatal glutamate release, enhanced striatopallidal GABA release and motor deficits in PD --- p.154 / Chapter 6.2 --- Enhanced prelimbic cortical glutamate release onto accumbal D2 MSNs and cognitive deficits in PD --- p.155 / Abbreviations --- p.158 / References --- p.160

At the neuronal synapse, neurotransmitter-filled synaptic vesicles (SVs) fuse with the presynaptic plasma membrane during activity. Following exocytosis, SVs must be retrieved for neurotransmission to be maintained. Several modes of SV recycling have been identified. During mild neuronal activity, clathrin-mediated endocytosis has been regarded as the dominant SV retrieval mode, however the recently identified ultrafast endocytosis mode may also be important in this condition. During elevated activity, activity-dependent bulk endocytosis (ADBE) is the dominant SV retrieval pathway. In ADBE, large invaginations are formed from the plasma membrane, which then undergo scission to create bulk endosomes. In a second distinct step, SVs bud from these endosomes and specifically repopulate the reserve SV pool. However, since its first identification, only few molecules have been shown to participate in ADBE. The aim of this PhD was to identify novel molecules and elucidate the molecular mechanism of ADBE. To achieve this, two independent biochemical approaches were designed to purify and enrich bulk endosomes from primary neuronal cultures. In the first approach, bulk endosomes and SVs were labelled with a dye, FM1-43, using a strong stimulus. Cells were broken mechanically and the post nuclear supernatant, that contains all intracellular organelles, was collected. The supernatant was then subjected to subcellular fractionation using discontinuous Nycodenz gradients. This stimulated sample was always processed in parallel with a basal sample, where no neuronal stimulus was applied, in order to visualise activity dependent FM loading. After different fractionation protocols were applied, bulk endosomes were efficiently separated from SVs, as revealed by tracking fluorescence in different fractions. The fractionation results were further validated by electron microscopy, where bulk endosomes and SVs were labelled with horseradish peroxidase and purified using the established protocol. Immunoblotting against selected SV cargo proteins from stimulated bulk endosome and SV samples, indicated the specific and preferential localisation of VAMP4 on bulk endosomes, in contrast to other SV cargo. The molecular identity of bulk endosomes was also approached by submitting the bulk endosome fractions to semi-quantitative mass spectrometry. This analysis revealed many different proteins that were identified in bulk endosome samples and quantification approaches further indicated proteins that can be localised on bulk endosomes and have a potential role in ADBE. A second magnetic isolation approach was designed, to purify bulk endosomes using a completely different methodology. In this case, bulk endosomes were specifically labelled with iron nanoparticles, which are preferentially taken up by bulk endosomes since they are larger than SVs. The cells were broken as before and post nuclear supernatant was acquired. In this case, the supernatant was submitted to magnetic isolation that separated iron beads labelled structures from all other intracellular organelles. An extensive immunoblotting analysis of magnetic bulk endosomes validated that VAMP4 and syndapin I, two essential ADBE proteins, were enriched in these purified samples. These magnetic bulk endosomes were also analysed using semi-quantitative MS and revealed many proteins with a potential role in ADBE. Significant overlap between the two independent methods was observed, further validating these approaches. Combining these two methods with bioinformatics tools allowed the identification of the molecular signature of ADBE as well as novel key candidates for this process. Specific molecules were investigated for their role in ADBE and SV recycling using a variety of different real-time fluorescent imaging assays. A major focus was on rab small GTPases. High molecular weight dextran uptake was used to specifically study the role of these proteins in ADBE, as it preferentially reports uptake via larger bulk endosomes. A pH sensitive chimeric protein, synaptophysin-pHluorin, was used to investigate the role of these proteins in CME. Additional imaging assays were used to answer emerging questions regarding the function and localisation of these targets in the presynapse. Using these approaches, rab11A and rab35 were found to promote ADBE and accelerate clathrin-mediated endocytosis. This effect was specific to high intensity stimulation, while SV exocytosis was not affected. Further research on the role of both novel and established ADBE molecules will provide key future insights into the mechanism of both bulk endosome generation/scission and subsequent SV reformation. A very promising group is rab proteins and now evidence for their implication in SV recycling is presented here. Identification and characterisation of new targets will allow to investigate the role of ADBE in neurotransmission in both physiology and pathophysiology.

A ivermectina é uma lactona macrocíclica utilizada para o tratamento de parasitoses na espécie humana e amplamente empregada em medicina veterinária como endectocida. Em mamíferos, diversas evidências indicam que as lactonas macrocíclicas interagem com canais de cloro mediados pelo ácido gama-aminobutírico (GABA). Sabe-se que o sistema GABAérgico está envolvido com a manifestação do comportamento sexual. Assim, no presente trabalho foram estudados, em ratos, os efeitos da ivermectina na coordenação motora, na motivação e no desempenho sexual de machos, bem como no comportamento sexual de fêmeas; além disso, avaliaram-se os níveis séricos de testosterona e as concentrações de diferentes neurotransmissores e seus metabólitos no hipotálamo e estriado, áreas do sistema nervoso central relacionadas, respectivamente, com o comportamento sexual e atividade motora. No presente estudo foram usadas as doses de 0,2 e 1,0 mg/kg, por via intraperitoneal, de uma formulação comercial de ivermectina; estas doses foram escolhidas considerando que a primeira é a dose usualmente empregada terapeuticamente e a segunda baseada em achados prévios que mostraram interferência no comportamento sexual de ratos machos. Os resultados mostraram que a administração de ivermectina em ratos: promoveu prejuízo na coordenação motora de machos; não interferiu na motivação sexual e na ereção peniana de machos; prejudicou o comportamento sexual de ratas; não alterou o peso relativo dos testículos, epidídimos, próstata e vesícula seminal, porém aumentou o peso relativo do fígado; não alterou o índice gonadossomático de machos, bem como não foram observadas alterações no estudo histopatológico dos diferentes órgãos; reduziu os níveis séricos de testosterona de machos; diminuiu os níveis de GABA hipotalâmico e estriatal, bem como reduziu a atividade dos sistemas serotoninérgico e dopaminérgico hipotalâmicos e, no estriado, reduziu a atividade do sistema dopaminérgico. Os resultados foram discutidos considerando a interferência da ivermectina nos sistemas de neurotransmissores central. / Ivermectin is a macrocyclic lactone used for the treatment of parasitic infections in humans and widely used in veterinary medicine as endectocide. In mammals, a number of evidence indicate that interact with the macrocyclic lactones chloride channels mediated by gamma-aminobutyric acid (GABA). It is known that the GABAergic system is involved in the manifestation of sexual behavior. Thus, in the present work were studied in rats, the effects of ivermectin on motor coordination, motivation and sexual performance of males and females in sexual behavior; furthermore, evaluated the serum levels of testosterone and different concentrations of neurotransmitters and their metabolites in the hypothalamus and striatum areas of the central nervous system related, respectively, sexual behavior and motor activity. In the present study doses of 0.2 and 1.0 mg / kg were used, intraperitoneally, a commercial formulation of ivermectin; these doses were chosen considering that the first dose is usually employed therapeutically and the second based on previous findings have shown that interference with sexual behavior of male rats. The results showed that administration of ivermectin in rats: impairment in motor coordination promoted male; did not interfere on the sexual motivation and the penile erection in male; impaired sexual behavior of female rats; did not alter the relative weight of the testes, epididymis, prostate and seminal vesicles, but increased the relative liver weight; did not alter the gonadosomatic index (GSI) of males and no changes were observed in the histopathological study of different organs; reduced serum testosterone levels of male; decreased levels of hypothalamic and striatal GABA and reduced the activity of the serotonergic and dopaminergic systems hypothalamic and the, reduced the activity of the dopaminergic system striatal. The results were discussed considering the interference of ivermectin in central neurotransmission systems.

O glifosato é um herbicida utilizado na agricultura para o controle de plantas daninhas. No Brasil o glifosato é comercializado com vários nomes, dentre eles Roundup®; esses produtos são acrescidos de ingredientes ditos inertes que permitem a penetração do glifosato através da membrana plasmática da planta, potencializando a sua ação, além de propiciar maior estabilidade e potencial de bioacumulação. Considerando que a presença de certas substâncias químicas em formulações comerciais de glifosato pode aumentar sua toxicidade e que há carência de estudos sobre a toxicologia do desenvolvimento com essas formulações, o presente estudo investigou os efeitos da exposição perinatal de ratos ao Roundup®, a fim de avaliar o comportamento e neuroquímica cerebral materna, bem como o desenvolvimento físico e neurocomportamental da prole, desde o nascimento até a idade adulta. Para tanto, ratas prenhes receberam 50, 100 e 150 mg/kg do 15° dia de gestação até o 7° dia da lactação. Os resultados mostraram que o glifosato- Roundup® não alterou o peso materno durante os períodos de exposição ao herbicida e tampouco o consumo de água e ração das ratas, sugerindo ausência de toxicidade materna. Por outro lado, observou-se que o número de filhotes vivos de todos os grupos de ratas expostas ao herbicida foi menor do que aquele das fêmeas do grupo controle. Nas ratas expostas ao glifosato-Roundup® observou-se redução de atividade geral no campo aberto na maior dose, prejuízo no comportamento materno (CM) e no comportamento materno agressivo (CMA), bem como no hipotálamo aumento da atividade do sistema dopaminérgico e redução da atividade do sistema serotoninérgico, enquanto no hipocampo e estriato houve diminuição da atividade do sistema dopaminérgico. Esses achados sugerem que o aumento da atividade do sistema dopaminérgico hipotalâmico, por reduzir a liberação de prolactina, um hormônio essencial para a expressão do CM, prejudicou a manifestação desse comportamento. No mesmo sentido, a serotonina e dopamina por modularem os efeitos da ocitocina, um peptídeo importante no controle e na eliciação do CM e CMA, podem ter contribuído para os presentes achados. Quanto ao estudo dos parâmetros de desenvolvimento físico e neurocomportamental da prole, não foram modificados pela administração do herbicida. No presente trabalho foi avaliada também a atividade geral da prole na infância e na idade adulta, mostrando que as alterações foram pontuais tanto na prole masculina como na feminina, não caracterizando alterações motoras ou emocionais. Quanto ao comportamento de brincar de luta, a administração do glifosato-Roundup® reduziu apenas o pinning tanto de ratos machos como de fêmeas, sem alterar outros parâmetros deste comportamento. A duração e frequência do pinning são consideradas como parâmetros que expressam o início do comportamento social do rato. Desta forma, nestes animais não foram estabelecidos os padrões de comportamento social durante o desenvolvimento, porém aqueles relacionados com o comportamento de luta e sexual foram preservados. Na idade adulta, não foram observadas alterações na interação social de ratos e ratas expostos ao herbicida, ou seja, não promoveu efeitos em longo prazo no comportamento social. Quanto aos achados de neuroquímica cerebral da prole adulta exposta ao herbicida, foram encontradas alterações nos diferentes sistemas de neurotransmissão, contudo a ausência de alterações relevantes na atividade geral e na interação social das proles masculina e feminina adultas, não foi possível especular sobre o envolvimento desses sistemas de neurotransmissão com esses comportamentos. Esses achados em conjunto, mostraram que a exposição perinatal a diferentes doses de glifosato- Roundup® promoveu prejuízo no comportamento materno, os quais foram associados aos sistemas de neurotrnasmissão cerebrais, e na prole, embora não tenham sido observados efeitos no desenvolvimento físico e neurocomportamental, ocorreram alterações comportamentais na infância que se estenderam até a idade adulta. / Glyphosate is a herbicide used in agriculture to control weeds. In Brazil glyphosate is marketed under several names, including Roundup ®. Ingredients considered inert are added to these products in order to allow the penetration of glyphosate across the plasma membrane of the plant, increasing its action, as well as providing greater stability and bioaccumulation potential. Considering that the presence of certain chemicals in commercial formulations of glyphosate may increase its toxicity and that there are insufficient data on the toxicology of development using these formulations, the present study investigated the effects of perinatal exposure of rats to Roundup®, to assess behavior and maternal brain neurochemistry, as well as physical and neurobehavioral development of the offspring from birth to adulthood. To this end, pregnant rats received 50, 100 and 150 mg/kg from the 15th gestation day to the 7th lactation day. The results showed that glyphosate-Roundup® did not alter maternal weight during neither periods of exposure to the herbicide nor the consumption of water and feed the rats, suggesting the absence of maternal toxicity. On the other hand, it was observed that the number of living offspring of all groups of rats exposed to herbicide was smaller than that of the female control group. In rats exposed to glyphosate-Roundup® it was possible to observe a reduction in general activity in the open field at the highest dose, impairment in maternal behavior (CM) and maternal aggressive behavior (CMA), as well as in the hypothalamus there was an increase in the dopaminergic system activity and a reduction of the serotonergic system activity, whereas the hippocampus and striatum showed decreased activity of the dopaminergic system. These findings suggest that the increased activity of the hypothalamic dopaminergic system by reducing the release of prolactin, an essential hormone for the expression of CM, damaged the manifestation of this behavior. Similarly, serotonin and dopamine, which modulate the effects of oxytocin, an important peptide in the control and elicitation of the CM and CMA, may have contributed to the present findings. Regarding the study of the parameters of physical and neurobehavioral offsprings development, no modification was caused by the administration of the herbicide. The present study also evaluated the overall activity of the offspring in childhood and adulthood, showing that the changes were punctual in both male and female offspring in not characterizing motor or emotional changes. Regarding the behavior of play fighting, the presence of glyphosate-Roundup® reduced only the pinning from both male and female rats, without changing other parameters of this behavior. The duration and frequency of pinning are considered parameters which express the beginning of the social behavior of the rats. Thus, during the development of these animals, the patterns of social behavior have not been established, but those related to fighting and sex behavior were preserved. In adulthood, no changes were observed in the social interaction of male and female rats exposed to the herbicide, i. e., it did not promote long-term effects on social behavior. In the present study, considering the findings of brain neurochemistry of adult offspring exposed to the herbicide, it was possible to observe in the hypothalamus an increase in dopamine turnover in both male and female adult offspring and an increased turnover of serotonin only in male offspring. In the hippocampus, the most relevant findings were increased turnover of serotonin in males and in females the opposite effect. In the striatum an increased turnover of serotonin was found in both males and females, and dopamine only in females. Taking these data together and considering the absence of material changes in general activity measured in the open field and social interaction of male and female adult offspring, it is not possible to speculate about the relation of these neurotransmitter systems with these behaviors.

Macrophages and other cells of the innate immune system recognize foreign particles that could be potentially dangerous and respond by initiating an inflammatory response. The biologically active chemical mediators of this response called pro-inflammatory cytokines are produced in various myeloid derived immune cells and can affect other cells of the body. Interleukin-1β, a pro-inflammatory cytokine, has been shown to have direct effects on dorsal root ganglion (DRG) cell bodies including the upregulation and direct release of a nociceptive neurotransmitter called substance P (SP). Using a zymosan-induced model of systemic inflammation, we hypothesized that murine DRG neurons and the nerve processes associated with them in the dorsal horn of the spinal cord (SC) at the L1 level will show an upregulation of SP expression in response to inflammation in the peritoneum. Experimental mice were treated with a zymosan suspension (500mg/kg, intraperitoneal injection), and control mice received sterile filtered solution (intraperitoneal injection). Both DRG and SC specimens were collected after in situ fixation and subjected to immunofluorescence staining to label SP. Using confocal microscopy, fluorescence microscopy, and image analysis software this expression of SP was quantified and compared. In both tissue specimen groups, an increase in SP expression was discovered in zymosan treated mice. The exact cause of this increase was not specifically determined in this experiment. This experiment provided valuable insight about how a systemic inflammatory response can affect sensory nerve function. Successful methods for further experimentation were identified and information about the zymosan model of inflammation was obtained