Regulatory mechanisms of acetylcholine synthesis and release by T cells
Regulatory mechanisms of acetylcholine synthesis and release by T cells
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DOI：
10.1016/j.lfs.2012.04.031
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发表时间：
2012-11-27
期刊：
影响因子：
6.1
通讯作者：
Kawashima, Koichiro
中科院分区：
文献类型：
--
作者：
Fujii, Takeshi;Takada-Takatori, Yuki;Kawashima, Koichiro
目的：毒蕈碱和烟碱乙酰胆碱 (ACh) 受体在免疫细胞中表达。由胆碱乙酰转移酶 (ChAT) 合成并在 T 细胞中释放的 ACh 与这些受体结合。此外，我们最近证明了介导细胞（液泡 H+-ATP 酶的 16 kDa 蛋白脂质亚基的同源寡聚物）参与 T 细胞的 ACh 释放。在本研究中，我们研究了佛波醇12-肉豆蔻酸酯13-乙酸酯（PMA）、二丁酰cAMP（dbcAMP）和免疫抑制剂钙调神经磷酸酶抑制剂FK506对T细胞淋巴细胞胆碱能活性的影响。主要方法：我们采用灵敏、特异的方法测定了人白血病T细胞系MOLT-3细胞中ACh的含量和释放。 乙酰胆碱放射免疫测定法。此外，分别使用逆转录聚合酶链反应和 Fonnum 方法研究了 ChAT mRNA 的表达和 ChAT 活性。 主要发现：植物血凝素 (PHA)，一种 T 细胞激活剂，上调 ChAT mRNA 表达、ACh 的合成和释放。 PMA（一种蛋白激酶 C (PKC) 激活剂）和 dbcAMP（一种蛋白激酶 A (PKA) 激活剂）也通过上调 ChAT 基因表达来增加 ChAT 活性和 ACh 合成。 FK506 抑制 PHA 诱导的 ChAT mRNA 表达上调，表明钙调磷酸酶介导的途径参与 ChAT 基因转录。意义：PKC 和 PICA 的激活上调 T 细胞中的 ACh 合成，免疫激活通过钙调磷酸酶介导的途径触发 ChAT 基因转录。 (C) 2012 Elsevier Inc. 保留所有权利。
Aims: 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.Main methods: 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.Key findings: 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.Significance: Activation of PKC and PICA up-regulates ACh synthesis in T cells, and immunological activation triggers ChAT gene transcription through calcineurin-mediated pathways. (C) 2012 Elsevier Inc. All rights reserved.