Regulation of Mechanisms underlying signal transduction via sigma-1 receptor and activity of Tyrosine Hydroxylase.

通过 sigma-1 受体和酪氨酸羟化酶活性调节信号转导的机制。

基本信息

批准号：
09680781
负责人：
YAMAMOTO Hideko
金额：
$ 2.11万
依托单位：
Tokyo Institute of Psychiatry
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (C)
财政年份：
1997
资助国家：
日本
起止时间：
1997 至 1999
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-09680781/
关键词：
Tyrosine Hydroxylase V-1 NO sigma-1 receptors c-jun dopamine beta-hydroxylase mutation primary cultured neuronal cells GST融合蛋白質トランスジェニックマウスノルアドレナリンアドレナリン会合蛋白質 Xenopus oocyte 転写因子 CREB

项目摘要

(1). Mechanism underlying signal transduction via intracelluar sigma-1 receptors. Expression cloning of sigma-1 receptor (sR1) in Xenopus oocytes showed a binding ability for NE-100, a ligand of sR1, as observed in native tissues. Site-directed mutagenesis in the putative transmembrane (TN) domain of sR1 did not alter expression levels of mutant sR1. [ィイD13ィエD1H]NE-100 binding was abolished by the double mutation (S99! and LL105-106AA) or by the single mutation (Y103F). Scatchard analysis using [ィイD13ィエD1H](+)pentazocine as a ligand demonstrated that the single mutation (Y103F) resulted in low affinity, however, the double mutation (S99A, LL105-106AA) had no effect. These findings provide evidence that the putative TM domain of sR1 plays a critical role in ligand-binding.(2). Kinetic Characterization of the nitric oxide (NO) toxicity for PC12 cells: effects of half-life time of NO release. Effects of low concentrations of nitric oxide (NO) on cell viability using NO donors. Exposure of … More undifferentiated PC12 cells to low concentrations of NO donors resulted in cell death in a dose- and time-dependent manner. The NO-induced cell death was characterized as apoptosis-like cell death. Pretreatment with an antioxidant ascorbic acid completely prevented the cell death. These findings suggest that the cell toxicity induced by NO at low concentrations strongly depends upon the duration of expose to NO from NO-donors.(3). A novel protein containing cdc 10/SWI6 motifs regulates expression of mRNA encoding catecholamine biosynthesizing enzymes. Mechanisms that control expression of catecholaminergic biosynthesizing enzymes in a transmitter phenotype-specific manner, are poorly understood. We provide evidence that overexpression of a novel cdc10SWI6 motif-containing protein, V-1, elicits the coordinate up-regulation of above enzymes in PC12D, and catecholamine levels are increased. Furthermore, V-1 is strongly expressed in the cytoplasm of rat chromaffin cells of adrenal medulla. Thus, V-1 may act as a cytoplasmic protein/protein adapter and be involved in control of the catecholaminergic phenotype expression via an intracellular pathway signaling to the nucleus. Less

(1). 非洲爪蟾卵母细胞中 sigma-1 受体 (sR1) 的表达克隆显示出对 sR1 配体 NE-100 的结合能力，如在天然组织中所观察到的。 sR1 假定的跨膜 (TN) 结构域中的定向诱变不会改变突变体 sR1 的表达水平。 [D13D1H]NE-100 结合被双突变（S99！和 LL105-106AA）或单突变（Y103F）消除，使用 [D13D1H](+)喷他辛作为配体的斯卡查德分析证明单突变（Y103F）。）导致亲和力低，然而，双突变（S99A， LL105-106AA) 没有影响。这些发现证明 sR1 的推定 TM 结构域在配体结合中发挥着关键作用。(2)。 NO 释放的寿命。使用 NO 供体的低浓度一氧化氮 (NO) 对细胞活力的影响将未分化的 PC12 细胞暴露于低浓度的 NO 供体中导致细胞死亡。以剂量和时间依赖性方式死亡。NO诱导的细胞死亡被描述为用抗氧化剂抗坏血酸预处理完全阻止了细胞死亡。浓度取决于暴露于 NO 供体的 NO 的持续时间。(3). 含有 CDC 10/SWI6 基序的新型蛋白质可调节编码儿茶酚胺生物合成酶表达的 mRNA 的表达。我们对以递质表型特异性方式产生的儿茶酚胺能生物合成酶知之甚少，我们提供的证据表明，一种新型 cdc10SWI6 基序蛋白 V-1 的过度表达会引起 PC12D 中上述酶的协调上调，并且儿茶酚胺水平增加。此外，V-1 在大鼠肾上腺髓质嗜铬细胞的细胞质中强烈表达，因此，V-1 可能充当一种受体。细胞质蛋白/蛋白接头，并通过向细胞核发出信号的细胞内途径参与儿茶酚胺能表型表达的控制。