Repeated stress has been reported to cause reversible impairment to the hippocampus. Glutamatergic and cholinergic systems were proposed to be involved in responses seen after exposure to stress and cholinesterase inhibitors. Effects of concurrent exposure to repeated stress and chlorpyrifos (CPF) on concentrations of excitatory amino acids, activities of cholinergic enzymes, and maximum binding density (Bmax) and equilibrium dissociation rate constant (Kd) of NMDA and total muscarinic receptors were studied in Long-Evans rats. The study was divided into 4 experiments. The first experiment was to find the dose of CPF to use for studies on the interaction of stress and CPF. From the results obtained, 60% of the maximum tolerated dose was chosen. An experiment to determine effects of repeated stress and CPF on cholinergic enzymes and glutamate included groups of rats (n=7-8) that were handled 5 days/week; restrained 1 hour/day for 5 days/week; swum 30 minutes for 1 day/week; or restrained 4 days/week and swum for 1 day/week, for 28 days. On day 24, each group was injected either with corn oil or CPF 160 mg/kg sc 4 hours after restraint. On day 28, blood samples were collected for acetylcholinesterase (AChE) activity. Brains were dissected into hippocampus (HP) and cerebral cortex (CC) to determine activities of acetylcholinesterase (AChE), carboxylesterase (Cbxy), and choline acetyltransferase (ChAT), and glutamate and aspartate concentrations. CPF inhibited AChE activity in blood, CC and HP, but stress did not affect AChE activity. Repeated restraint with swim reduced Cbxy and CPF inhibited Cbxy. Restraint with swim had a statistical trend to increase concentrations of glutamate in the HP more than swim alone (p = 0.064); but CPF had no effect on glutamate in the HP. CPF decreased concentrations of elevated aspartate in the HP of rats that were restrained and swum. The results suggested that restraint with swim indirectly elevated acetylcholine in the CC, and tended to increase glutamate in the HP. The experiment designed to study the effects of concurrent exposure to stress and CPF on NMDA and total muscarinic receptors was designed similar to the previous study, except that endpoints were Bmax and Kd of NMDA and total muscarinic receptors in the HP and CC, and NMDA receptors in the hypothalamus (HT). Restrained rats had higher Kd of NMDA receptors in the HP than control and restrained with swim rats; however, Bmax was similar. CPF deceased Bmax and Kd of total muscarinic receptors in the CC of swum rats (237.64 &#177; 17.36 fmol/mg protein, 0.216 &#177; 0.023 nM) and CPF also decreased Bmax of total muscarinic receptors in the CC of restrained rats (229.08 &#177; 17.36 fmol/mg protein). There were no effects of stress, CPF, or interactions of stress and CPF on NMDA receptors in the CC or on total muscarinic receptors in the HP. In summary, CPF was capable of modulation of total muscarinic receptors of swum and restrained rats, suggesting that cholinergic transmission in the CC for cognition, sensory and motor activity may be modified. Furthermore, we examined effects of stress and CPF on concentrations of monoamines. Swim stress and CPF individually decreased concentrations of norepinephrine in the HP, whereas swim and restraint with swim decreased concentrations of norepinephrine but increased concentrations of DOPAC in the HT. Swim stress increased concentrations of dopamine in the HT more than control or restraint. CPF did not alter concentrations of norepinephrine, dopamine, or DOPAC in the HT. The interactions of repeated stress and CPF on serotonin approached significance in the HP (p = 0.06) and HT (p = 0.08). CPF increased serotonin concentrations in rats that were handled and restrained but not swum. CPF reduced the elevated concentrations of serotonin in restrained rats and restrained with swim rats (p < 0.05). Swim and restraint with swim were potential stress models that altered noradrenergic, dopaminergic, and serotonergic responses in the HT. In summary, repeated stressors had effects on glutamatergic, cholinergic, and monoamine systems. CPF had effects on cholinergic and monoamine systems but the interactions between stress and CPF were few. / Ph. D.

The nervous system and immune system have broad and overlapping distributions in the body, and interactions of these ubiquitous systems are central to the field of neuroimmunology. Over the past two decades, there has been explosive growth in our understanding of neuroanatomical, cellular, and molecular mechanisms that mediate central modulation of immune functions through the autonomic nervous system. A major catalyst for growth in this field was the discovery that vagal nerve stimulation (VNS) caused a prominent attenuation of the systemic inflammatory response evoked by endotoxin in experimental animals. This effect was mediated by acetylcholine (ACh) stimulation of nicotinic receptors on splenic macrophages. Hence, the circuit was dubbed the “cholinergic anti-inflammatory pathway”. Subsequent work identified the α7 nicotinic ACh receptor (α7nAChR) as the crucial target for attenuation of pro-inflammatory cytokine release from macrophages and dendritic cells. Further investigation made the important discovery that cholinergic T cells within the spleen and not cholinergic nerve cells were the source of ACh that stimulated α7 receptors on splenic macrophages. Given the important role that inflammation plays in numerous disease processes, cholinergic anti-inflammatory mechanisms are under intensive investigation from a basic science perspective and in translational studies of animal models of diseases such as inflammatory bowel disease and rheumatoid arthritis. This basic work has already fostered several clinical trials examining the efficacy of VNS and cholinergic therapeutics in human inflammatory diseases. This review provides an overview of basic and translational aspects of the cholinergic anti-inflammatory response and relevant pharmacology of drugs acting at the α7nAChR.

Diss. (sammanfattning) Stockholm : Karol. inst., 2005. / Härtill 4 uppsatser.

published_or_final_version / Physiology / Doctoral / Doctor of Philosophy

Kateřina Janská Evaluation of the kinetics of acetylcholinesterase inhibitors in vitro Diploma thesis Charles University in Prag, Faculty of Pharmacy in Hradec Králové Pharmacy Department of Biological and Medical Sciences Supervisor: Doc. MUDr. Josef Herink, DrSc. Consultant: PharmDr. Vendula Šepsová The aim of the thesis was to determine the type of an inhibition of newly synthesized AChEI and to find out if AChEI structure changes influence the type of an inhibition. Altogether 12 substances (7 tacrine hybrides and 5 7-methoxy- donepezil hybrids) were investigated. The inhibition potential of the tested substances was studied in vitro on the human recombinant AChE. Spectrophotometric Ellman method was utilized as the measurement tool. The noncompetitive type of an inhibition for substances EN 1-5, PC-25 and PC-33, mixed type of an inhibition for substances PC-48 and PC-49, uncompetitive type of an inhibition for substances EN-6, EN-7 and competitive type of an inhibition for the substance PC-37 was determined. The greatest inhibition potential according to Ki values were found for substances EN-7 and PC-37. Substances PC-37 and PC-48 were determined as substances with the biggest affinity to the AChE. The type of an inhibition has been influenced by a substituent position in PC substances and by...

Adéla Dolejšová Testing the efficacy of potential therapeutics for Alzheimer,s disease Charles University in Prague, Faculty of Pharmacy in Hradec Kralove Pharmacy The aim of this thesis was to find out whether already implemented Ellman's method, which is used to analyse irreversible inhibitors AChE, is going to be a suitable technique for measuring reversible inhibitors. Furthermore, the efficiency of newly synthesized AChE inhibitors was established. These inhibitors will be used for treating AD or as prophylaxis against neural paralytic substances. Moreover, it was compared affect the efficacy of AChE inhibitors after intramuscular and intraperitoneal administration. The measurements were done in vivo on potkan species Wistar. In the first experiment standard AChE (tacrin, 7-MEOTA, donepezil, rivastigmin) inhibitors were applied to verify the method. In the second experiment the newly synthesizes AChE (K 298, K 344, K 474) inhibitors were medicine. The results confirmed the effectiveness of the tested method on commonly used inhibitors such as donepezil and rivastigmin. Out of the newly synthesized K 298, K 344 and K 474 inhibitors none was proven to have any significant inhibiting activity. Key words: acetylcholinesterase, acetylcholinesterase inhibitors, Alzheimer disease, cholinergic system,...

<div> <p>Fluorescent proteins and the biosensors created with them have been used extensively to monitor chemical species inside and outside of the cell. They have been used to increase our knowledge of cellular function in normal and diseased states. Fluorescent biosensors are advantageous because they can be genetically encoded, do not require exogenous reagents, and can be quantitative. Fluorescent biosensors are also able to measure analytes with high spatial and temporal resolutions, enabling measurements at the scale of physiological events. In this thesis efforts have made to increase the available fluorescent biosensor tools for imaging cellular events. This work includes creation of new sensors for two molecules not yet detectable via fluorescent protein biosensor, acetylcholine and adenosine diphosphate. Efforts were also made to improve the current available biosensors for adenosine triphosphate and cellular redox, to make them more compatible with multiplex and deep tissue imaging. Here I present my work to design, characterize and utilize these fluorescent biosensors.</p> </div> <br>

A farmacodependência é considerada uma doença crônica e sujeita à recaídas, na qual o indivíduo perde o controle sob a utilização de determinada droga de abuso. Conforme o usuário persiste com o uso da droga, ocorrem alterações anatômicas, fisiológicas e neuroquímicas no sistema nervoso central (SNC), as quais podem culminar no desenvolvimento de um comportamento compulsivo. A neurobiologia deste processo é complexa e envolve mecanismos de plasticidade em diferentes sistemas neurotransmissores. O principal deles é o sistema mesocorticolímbico dopaminérgico, constituído por neurônios da área ventral do tegmento mesencefálico (VTA) que se projetam para o núcleo accumbens (NAc) e ao córtex pré-frontal (CPF), diretamente relacionado aos processos motivação e recompensa. Contudo, o mesmo não é suficiente para elucidar a complexidade da doença, o que levou ao entendimento da presença de outros sistemas neurotransmissores neste processo. Sabe-se que o sistema colinérgico muscarínico está diretamente envolvido em diferentes doenças neuropsiquiátricas, incluindo a farmacodependência. Além disso, os receptores colinérgicos muscarínicos (mAChRs) estão densamente presentes em regiões límbicas, onde acetilcolina e dopamina interagem por neuromodulação. Diante disto, o objetivo deste trabalho foi investigar as possíveis alterações plásticas no sistema colinérgico muscarínico resultantes de tratamentos com cocaína que mimetizaram o consumo compulsivo humano. Para tanto, foram realizados ensaios comportamentais com camundongos Swiss machos adultos em campo aberto, tratados durante um (acute binge paradigm, 30 mg/kg) ou 14 dias (escalating dose binge paradigm, 15 - 30 mg/kg) com cocaína. Os animais receberam 3 injeções intraperitoneais (i.p.) de cocaína com intervalos de 60 minutos, durante os quais a atividade locomotora foi avaliada. Após a análise comportamental, os animais foram eutanasiados por decapitação para a remoção do encéfalo e dissecação do estriado, CPF e hipocampo, regiões cerebrais cruciais para o processo fisiopatológico da farmacodependência. Componentes do sistema dopaminérgico (receptores D1 e D2) e colinérgico muscarínico (M1-M5 mAChRs, ChAT, VAChT e AChE) foram avaliados por Immunoblotting. O sangue dos animais foi coletado para a realização das dosagens de cocaína e benzoeilecgonidina por UPLC-MS/MS. O desempenho locomotor total dos animais tratados com cocaína foi superior ao dos animais controle. O grupo tratado com escalonamento de dose desenvolveu sensibilização comportamental aos efeitos psicoestimulantes da cocaína no segundo dia de tratamento e, a partir dele, a atividade locomotora total manteve a mesma magnitude. Além disso, conforme o aumento da dose, os animais mantiveram um nível de atividade superior ao basal, mesmo após o término do experimento. As análises de Immunoblotting mostraram alterações dopaminérgicas e colinérgicas. No estriado observou-se redução da densidade de D2R após o tratamento de 14 dias e aumento na densidade de M3 mAChR após o tratamento agudo. Já no hipocampo observou-se redução de D1R e aumento de D2R, M1 e M5 mAChR após o tratamento crônico; e um aumento na densidade de M3 mAChR após o tratamento agudo. No CPF, foi evidenciada redução de M3 e de M5 mAChR após o tratamento cônico de 14 dias. Em relação às moléculas colinérgicas, observou-se, após o tratamento crônico, aumento da quantidade de ChAT em todas as estruturas estudadas. Além disso, VAChT mostrou-se aumentado no hipocampo após ambos os tratamentos. As dosagens plasmáticas revelaram a presença de 20,38 ± 3,4 ng/mL de cocaína e 224,6 ± 24,02 ng/mL de benzoilcgonina (BZE) nos animais do grupo agudo e, nos do grupo crônico, 62,26 ± 10,56 ng/mL e 375,1 ± 25,62 ng/mL de cocaína e BZE respectivamente. / Drug addiction is a chronic releapsing disorder characterized by the loss of control in limiting drug intake. As the drug use persists, anatomical, physiological and neurochemical changes occur in the central nervous system (CNS), which may lead to the development of compulsive behaviors. The neurobiology of this process is complex and involves mechanisms of plasticity in different neurotransmitter systems. The main one is the mesocorticolimbic dopaminergic system, composed by neurons from the ventral tegmental area (VTA) that projects to the nucleus accumbens (NAc), which is directly related to motivation and reward processes. However, just dopamine is not enough to elucidate the complexity of the disease, leading to the comprehension of another neurotransmitters system involved. It is known that the cholinergic system is involved in different neuropsychiatric disorders, including drug addiction. Furthermore, cholinergic muscarinic receptors (mAChRs) are densely present in limbic regions, where acetylcholine and dopamine interact by neuromodulation. Considering that, the aim of this study was to evaluate the existence of neuroadaptative changes in the cholinergic muscarinic system induced by cocaine in a compulsive-like behavior model in mice. Swiss-Webster adult male mice received 3 daily injections (i.p) of cocaine or saline, with a 60-min interval among them, either acutely (acute binge paradigm) or for 14 consecutive days (escalating dose binge paradigm). The locomotor activity was monitored in the open field during 60 min, in 5 min bins, after each injection. After behavioral analysis animals were euthanized by decapitation and the brain regions of striatum, hippocampus and prefrontal cortex, involved in the pathophysiology of addiction were dissected. Dopaminergic receptors (D1R and D2R), cholinergic muscarinic receptors (M1-M5 mAChRs), choline acetylytransferase (ChAT), acetylcholine vesicular transporter and acetylcholinesterase (AChE) were quantified by Immunoblotting. Blood samples were collected with heparin and plasma was separated and stored with 2% sodium fluorite at -80ºC for cocaine and benzoilecgonine quantification by UPLC-MS/MS. In the open field, animals treated with cocaine showed an increase in locomotor activity compared to control. Cocaine induced behavioral sensitization, in the escalating dose group on day 2, and after that the locomotor activity had the same magnitude until day 14th. These animals also kept the locomotor activity elevated even after the last injection. Immunobltting shows dopaminergic and cholinergic changes. An increase in M3 was observed in both hippocampus and striatum of animals acutely treated. After 14 days, there was an increase in M1, M5 and D2 and a decrease in D1 in hippocampus. There was also a decrease in D2 in the striatum; and finally, there was a decrease in M5 and M3 in the prefrontal cortex. ChAT densities were higher in all regions after the chronic treatment. Besides that, VAChT were higher in the hippocampus after both acute and chronic treatments. UPLC-MS/MS for cocaine and benzoilecgonine demonstrated the presence of 20,38 ± 3,4 ng/mL of cocaine and 224,6 ± 24,02 ng/mL of BZE in the acute binge group; and, 62,26 ± 10,56 ng/mL and 375,1 ± 25,62 ng/mL of cocaine and BZE, respectively in the escalating dose animals.

Introdução: Os pacientes com bronquiolite obliterante (BO) costumam apresentar acometimento importante da função pulmonar que resulta em hipoxemia crônica e limitação da atividade física. Não há terapêutica de grande eficácia na BO, a resposta aos broncodilatadores costuma ser pobre, entretanto, não se conhece a resposta broncodilatadora a um agente anticolinérgico de longa ação como o brometo de tiotrópio (BT). Já se demonstrou eficácia e segurança do BT em adultos com doença pulmonar obstrutiva crônica (DPOC) com resposta broncodilatadora significativa e sustentada em dose única diária. Objetivo: Avaliar se existe melhora do grau de obstrução brônquica e do aprisionamento aéreo, através de medidas funcionais, após o uso de dose única de brometo de tiotrópio por via inalatória comparado a placebo em crianças e adolescentes com BO. Métodos: Ensaio clínico prospectivo, duplo cego, randomizado, placebocontrolado e cruzado em pacientes com BO estáveis na faixa etária de 6 a 16 anos. Espirometrias e pletismografias foram realizadas antes e aos 30, 60, 120, 180 minutos e 24 horas após a inalação de 18 mcg de tiotrópio ou placebo. Após 7-14 dias, os medicamentos foram invertidos e os procedimentos repetidos. As mudanças nos parâmetros de função pulmonar em cada momento foram comparadas com o basal através da análise de variância (ANOVA) e pós-teste de Tukey e as diferenças entre todos os momentos versus o basal nos grupos tiotrópio versus placebo foram comparados usando o teste de Friedman. Resultados: Trinta pacientes participaram do estudo (23 do sexo masculino, 7 do feminino; idade 10,9±2,8a ), com valores basais de função pulmonar (% do previsto) de CVF, VEF1, VEF1/CVF, FEF25-75%, CI, CPT, VR, VR/CPT, resistência das vias aéreas (raw) e condutância (sGaw) de 75±15, 48±14, 59±11, 22±11, 64±23, 120±19, 281±101, 49±13, 250±65 e 23±9, respectivamente. Diferenças estatisticamente significantes foram observadas após inalação do tiotrópio nos seguintes parâmetros comparados ao tempo basal: CVF em 60/120/180 min/24h, VEF1 em 30/60/120/180min, VEF1/CVF em 60/120/180min, FEF25- 75% em 60/120/180min, VR em 30/60/120/180min, CPT em 30/120/180min, VR/CPT em 30/60/120/180min, raw em 30/60/120/180min/24h e sGaw em 30/60/120/180min/24h. Na fase placebo não houve diferença estatisticamente significante em nenhum parâmetro funcional em nenhum momento após a administração. As diferenças entre as medidas funcionais comparando o grupo tiotrópio versus o grupo placebo foram estatisticamente significantes. Conclusões: O brometo de tiotrópio, após dose única, diminuiu agudamente o grau de obstrução e de aprisionamento aéreo por até 24 horas em crianças com BO. Estudos de longo prazo são necessários para se avaliar o papel do BT na terapêutica desses pacientes / Introduction: Patients with bronchiolitis obliterans (BO) usually have severe airflow obstruction that results in chronic hypoxemia and limitation of physical activity. There is no efficient therapy for BO, bronchodilator response is usually poor, however, the bronchodilator response to a long action anticholinergic agent such as tiotropium bromide (TB) is not known. Efficacy and safety of TB with one daily administration has already been shown in chronic obstructive pulmonary disease (COPD) with significant and sustained bronchodilator response. Objective: Verify through functional measurements whether the level of bronchial obstruction and air trapping was improved by the administration of a single dose of TB by inhalation when compared to placebo in children and adolescents with BO. Methods: A randomized, double blind, placebo-controlled, crossover, prospective study in stable BO patients, 6 to 16 years of age. Spirometry and plethysmography were performed before and at 30, 60, 120 and 180 minutes and 24 hours after inhalation of 18 mcg of tiotropium or a placebo. After 7-14 days, the drugs were inverted, and the procedures were repeated. The changes in lung function parameters at each time point were compared to the baseline by analysis of variance (ANOVA) and Tukey\'s post-test and the differences in all time points assessment versus baseline in tiotropium versus placebo groups were compared using the Friedman test. Results: Thirty patients were enrolled in the study (23 male, 7 female; age 10.9±2.8 y) with baseline lung function values (% predicted) of FVC, FEV1, FEV1/FVC, FEF25-75%, IC, TLC, RV, RV/TLC, airway resistance (raw) and conductance (sGaw) of 75±15, 48±14, 59±11, 22±11, 64±23, 120±19, 281±101, 49±13, 250±65 and 23±9, respectively. Statistically significant differences were observed after tiotropium inhalation in the following parameters compared to baseline: FVC at 60/120/180min/24h, FEV1 at 30/60/120/180min, FEV1/FVC at 60/120/180min, FEF25-75% at 60/120/180min, RV at 30/60/120/180min, TLC at 30/120/180min, RV/TLC at 30/60/120/180min, raw at 30/60/120/180min/24h and sGaw at 30/60/120/180min/24h. For the placebo group, no significant differences were observed in any lung function parameters at any time. The differences between the functional measurements comparing the tiotropium versus placebo groups were statistically significant. Conclusions: Tiotropium bromide, after a single dose, acutely decreased airway obstruction and air trapping for up to 24 hours in children with BO. Long-term studies are necessary to evaluate the role of BT in the management of these patients

A resposta imune pode ser regulada tanto pelo SNS quanto pelo SNP. Estudos recentes têm identificado uma via colinérgica anti-inflamatória entre as fibras eferentes do nervo vago e direcionadas ao sistema imune. Desta forma, tem-se postulado que esta conexão provê um controle neural da inflamação aguda de uma forma direta e reflexa, sendo então chamada de reflexo inflamatório. Assim, pareceu-nos relevante estudar as influências do SNP na função das DCs, o que foi feito na vigência de um processo inflamatório do tipo antígeno específico produzido por OVA. Para tanto utilizamos: o Betanecol (agonista muscarínico), a Atropina (antagonista muscarínico), a Anabasina (agonista nicotínico) e a Mecamilamina (antagonista nicotínico). No presente trabalho observou-se que: A Anabasina aumentou a porcentagem de células que expressam as moléculas co-estimulatórias B7.1 e B7.2 no baço de camundongos e diminuiu a produção de IL-12 no sobrenadante de co-cultura de células de baço, enquanto que o Betanecol não produziu qualquer efeito no fenótipo das DCs e na produção de citocinas; A Mecamilamina e a Atropina não foram capazes de alterar o fenótipo de DCs de baço e nem a produção de citocinas numa co-cultura de células de baço. A Anabasina, por sua vez: diminuiu a expressão das moléculas co-estimulatórias B7.1 e B7.2 nas DCs de linfonodo; diminuiu a expressão de MHC-II e aumentou a expressão das moléculas co-estimulatórias B7.1 e B7.2 em DCs de baço; diminuiu a expressão de IL-12 intracelular e aumentou a expressão de NF-B de DCs de cultura de hepatócitos; diminuiu os níveis séricos de TNF e MCP-1 e aumentou os níveis séricos de IL-6; diminuiu a resposta de 10 hipersensibilidade do tipo tardia; aumentou a expressão de MHC-II, diminuiu a expressão das moléculas co-estimulatórias B7.1 e B7.2; diminuiu a produção de IL-12 e aumentou a produção de IL-10 nas DCs de medula óssea; aumentou a expressão de mNAChRa7 de DCs maduras provenientes de medula óssea. Os dados obtidos deste trabalho sugerem que o sistema colinérgico diminua a apresentação de antígenos específicos por DCs, atuando de forma anti-inflamatória e produzindo um shift da resposta Th1 para Th2; sugerem, ainda, que estes achados relacionam-se à estimulação da subunidade 7 do receptor nicotínico, com consequente aumento de atividade das DCs e subsequente aumento da expressão de mNAChRa7 / Immune responses might be regulated by the SNS and by PNS. Recently, it was shown the existence of a cholinergic anti-inflammatory pathway that connects vagus nerve afferent/efferent fibers to immune system cells within some organs. These connections make possible a neural control of the inflammatory response both throught a direct and reflex mechanism; the so called inflammatory reflex. Therefore, we thougth that it would be relevant to study the influences of PNS on DCs function in an antigen specific inflammatory process induced by OVA. As pharmacological tools: Bethanechol (muscarinic agonist), Atropine (muscarinic antagonist), Anabasine (nicotinic agonist) and Mecamylamine (nicotinic antagonist) were used. We showed that anabasine increased the percentage of splenocytes expressing co-stimulatory molecules (B7.1 and B7.2) and decreased IL-12p40 production in co-cultured (adherent:non-adherent) splenocytes supernatant. Bethanechol had no effects on DCs phenotype and cytokines production whatsoever. Mecamylamine and atropine also had no effects on DCs phenotype and on cytokines production, as well. Anabasine: decreased co-stimulatory molecules (B7.1 and B7.2) expression on DCs present in lymph nodes.Anabasine also decreased MHC-II expression, while increased the co-stimulatory molecules (B7.1 and B7.2) expression on DCs present in the spleen.Additionally, anabasine decreased intracellular IL-12 expression, while increased NF-B expression in splenocytes culture. Interestingly, anabasine decreased both TNF and MCP-1 and increased IL-6 serum levels. Anabasine also decreased a delayed type hypersensitivity (DTH) response in OVA-sensitized mice. Moreover, Anabasine increased both MHC-II expression and 12 IL-10 productions, while decreased both co-stimulatory molecules (B7.1 and B7.2) expression and IL-12 production in bone marrow derived DCs. Finnally, anabasine increased mNAChRa7 expression in mature bone marrow derived DCs. Taken together, these data suggest that the cholinergic system decreases antigen-specific presentation by DCs, leading to an anti-inflammatory effect, which in turn induces to a shift from Th1 to Th2 responses. Moreover, our data strongly suggest that nicotinic receptor 7 subunit is involved with PNS activity