Molecular Characterization of ADP-ribosyl Cyclase Coupled with Receptors

与受体偶联的 ADP-核糖基环化酶的分子表征

基本信息

批准号：
12480228
负责人：
HIGASHIDA Haruhiro
金额：
$ 8.45万
依托单位：
Kanazawa University
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
2000
资助国家：
日本
起止时间：
2000 至 2002
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-12480228/
关键词：
Ca signaling cyclic ADP-ribose Ca pool second messenger Signal transduction NAD ADPR cyclase cADPR サイクリックADPリボナース

项目摘要

We previously reported that stimulation of β-adrenergic and angiotensin II receptors modulates activity of ADP-ribosyl cyclase, a synthetic enzyme of cADP-ribose, and showed that cADP-ribose is a critical molecule in sympathetic potentiation of heart contraction, post-natal heart growth, and neuron-glia interaction. In the present study, we used the same strategy to explore the idea mat cADP-ribose is a second messenger downstream of the mGluRs. We first measured ADP-ribosyl cyclase activity in crude membranes from various rat and mouse nervous tissues in the presence or absence of glutamate and/or GTP. Second, the coupling preference of mGluRs to ADP-ribosyl cyclase was examined in NG108-15 neuroblastoma x glioma hybrid cells over-expressing each subtype.Glutamate stimulates ADP-ribosyl cyclase activity in rat or mouse crude membranes of retina via group III mGIuRs or in superior cervical ganglion via group I mGluRs. The retina of mGluR6 deficient mice showed no increase in the ADP-ribosyl cyclase level in response to glutamate. GTP enhanced the initial rate of basal and glutamate-stimutated cyclase activity. GTP-γ-S also stimulated basal activity. To determine whether the coupling mode of mGluRs to ADP-ribosyl cyciase is a feature common to individual cloned mGIuRs, we expressed each mGluR subtype in NG108-15 neuroblastoma x glioma hybrid cells. The glutamate-induced stimulation of the cyclase occurs preferentially in NG108-15 cells over-expressing mGluRs1, 3, 5, and 6. Cells expressing mGluR2 or mGluRs4 and 7 exhibit inhibition or no coupling, respectively. Glutamate-induced activation or inhibition of the cyclase activity was eliminated after pretreatment with cholera or pertussis toxin, respectively. Thus, the subtype-specific coupling of mGIuRs to ADP-ribosyl cyclase via G proteins suggests that some glutamate-evoked neuronal function is mediated by cADP-ribose.

我们之前报道过，刺激 β-肾上腺素能和血管紧张素 II 受体可调节 ADP-核糖基环化酶（cADP-核糖的合成酶）的活性，并表明 cADP-核糖是心脏收缩、产后心脏交感神经增强的关键分子。在本研究中，我们使用相同的策略来探索 cADP-核糖是神经元下游的第二信使的想法。我们首先测量了在存在或不存在谷氨酸和/或 GTP 的情况下来自各种大鼠和小鼠神经组织的粗膜中的 ADP-核糖基环化酶活性，其次，在 NG108-15 中检查了 mGluRs 与 ADP-核糖基环化酶的偶联偏好。神经母细胞瘤 x 神经胶质瘤杂交细胞过度表达每种亚型。谷氨酸刺激 ADP-核糖基环化酶活性在大鼠或小鼠的视网膜粗膜中，通过第 III 组 mGluR 或在上颈神经节中，通过第 I 组 mGluR。 mGluR6 缺陷小鼠的视网膜显示，谷氨酸对基础 GTP 的反应没有增加。和谷氨酸刺激的环化酶活性也刺激了mGluRs的基础活性。 ADP-核糖基环化酶是单个克隆 mGluR 的共同特征，我们在 NG108-15 神经母细胞瘤 x 神经胶质瘤杂交细胞中表达每种 mGluR 亚型，谷氨酸诱导的环化酶刺激优先发生在过表达 mGluRs1 的 NG108-15 细胞中，3。、5和6.表达mGluR2或的细胞mGluRs4和7分别表现出抑制或无偶联，谷氨酸诱导的环化酶活性的激活或抑制分别在用霍乱或百日咳毒素预处理后被消除，因此，mGluRs通过G蛋白与ADP-核糖基环化酶的亚型特异性偶联。表明一些谷氨酸诱发的神经元功能是由 cADP-核糖介导的。