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Regulatory mechanisms of acetylcholine synthesis and release by T cells.

机译:乙酰胆碱合成和T细胞释放的调节机制。

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Muscarinic and nicotinic acetylcholine (ACh) receptors are expressed in immune cells. ACh synthesized by choline acetyltransferase (ChAT) and released in T cells binds to these receptors. Furthermore, we have recently demonstrated the involvement of mediatophore, a homooligomer of a 16-kDa proteolipid subunit of vacuolar H(+)-ATPase, in ACh release from T cells. In this study, we investigated the effects of phorbol 12-myristate 13-acetate (PMA), dibutyryl cAMP (dbcAMP) and FK506, an immunosuppressant calcineurin inhibitor, on lymphocytic cholinergic activity in T cells.We determined the content and release of ACh in human leukemic T cell line MOLT-3 cells using a sensitive and specific radioimmunoassay for ACh. In addition, expression of ChAT mRNA and ChAT activity were investigated using reverse-transcription-polymerase chain reaction and Fonnum method, respectively.Phytohemagglutinin (PHA), a T-cell activator, up-regulated ChAT mRNA expression, synthesis and release of ACh. PMA, a protein kinase C (PKC) activator, and dbcAMP, a protein kinase A (PKA) activator, also increased ChAT activity and ACh synthesis by up-regulating ChAT gene expression. FK506 inhibited PHA-induced up-regulation of ChAT mRNA expression, suggesting the involvement of calcineurin-mediated pathways in ChAT gene transcription.Activation of PKC and PKA up-regulates ACh synthesis in T cells, and immunological activation triggers ChAT gene transcription through calcineurin-mediated pathways.
机译:肌肉蛋白和烟碱乙酰胆碱(ACH)受体在免疫细胞中表达。通过胆碱乙酰转移酶(聊天)合成,并在T细胞中释放与这些受体结合。此外,我们最近证明了来自T细胞的ACH释放的真空H(+) - ATP酶的16-KDA蛋白亚基的MOMOOLIGORER的参与。在这项研究中,我们研究了Phorbol 12-豆蔻酸酯13-乙酸酯(PMA),二丁酰磺醛蛋白(DBCAMP)和FK506,免疫抑制剂钙蛋白抑制剂在T细胞中淋巴细胞胆碱能活性的影响。我们确定了ACH中的含量和释放人白血病T细胞系Molt-3细胞使用敏感和特定的放射免疫测定用于ACH。此外,聊天mRNA的表达和ChAT活性使用逆转录 - 聚合酶链反应和Fonnum方法,respectively.Phytohemagglutinin(PHA),T细胞活化剂,上调ChAT的mRNA的表达,合成和乙酰胆碱释放的影响。 PMA,蛋白激酶C(PKC)活化剂和DBCAMP,一种蛋白激酶A(PKA)活化剂,也通过上调聊天基因表达增加了聊天活动和ACH合成。 FK506抑制PHA诱导的聊天mRNA表达的上调,表明钙突蛋白介导的途径在聊天基因转录中的累积。PKC和PKA up-Checlates在T细胞中的ACH合成,免疫激活触发通过钙素聊天聊天基因转录 - 介导的途径。

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