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一种在毒蕈碱胆碱能刺激下测量小鼠神经母细胞瘤细胞中鸟苷 3':5'-环磷酸相对变化的方法。

A method for measuring relative changes in guanosine 3':5'-cyclic monophosphate in mouse neuroblastoma cells on muscarinic cholinergic stimulation.

作者信息

Strange P G

出版信息

Biochem J. 1978 Nov 15;176(2):583-90. doi: 10.1042/bj1760583.

DOI:10.1042/bj1760583
PMID:217365
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC1186268/
Abstract

A convenient, inexpensive assay was developed for measuring relative changes in cyclic GMP in whole mouse neuroblastoma cells (clone NIE 115) based on labelling the cellular GTP pool with [8(-3)H]guanine. The time course of cell labelling and the distribution of radioactivity among possible products were studied; GTP is the only major labelled species. Radioactive cyclic GMP produced from the radioactive GTP on cell stimulation is isolated by column chromatography nad its identity has been rigorously established by paper chromatography and ion-exchange chromatography. The assay was used to study the time course of the cyclic GMP changes that occur after stimulation of neuroblastoma cells with carbamoylcholine and the dependence of the cyclic GMP changes on the carbamoylcholine concentration. The assay gives results comparable with those obtained by using a radioimmunoassay for cyclic GMP and should be applicable to other whole-cell and tissue-slice systems.

摘要

基于用[8(-3)H]鸟嘌呤标记细胞内GTP池，开发了一种简便、廉价的测定方法，用于测量全小鼠神经母细胞瘤细胞（克隆NIE 115）中环状GMP的相对变化。研究了细胞标记的时间进程以及放射性在可能产物中的分布；GTP是唯一主要的标记物种。细胞受刺激后，由放射性GTP产生的放射性环状GMP通过柱色谱法分离，其身份已通过纸色谱法和离子交换色谱法严格确定。该测定方法用于研究用氨甲酰胆碱刺激神经母细胞瘤细胞后环状GMP变化的时间进程以及环状GMP变化对氨甲酰胆碱浓度的依赖性。该测定方法给出的结果与使用环状GMP放射免疫测定法获得的结果相当，并且应该适用于其他全细胞和组织切片系统。

相似文献

1
A method for measuring relative changes in guanosine 3':5'-cyclic monophosphate in mouse neuroblastoma cells on muscarinic cholinergic stimulation.一种在毒蕈碱胆碱能刺激下测量小鼠神经母细胞瘤细胞中鸟苷 3':5'-环磷酸相对变化的方法。
Biochem J. 1978 Nov 15;176(2):583-90. doi: 10.1042/bj1760583.
2
Muscarinic responses and binding in a murine neuroblastoma clone (N1E-115). Selective loss with subculturing of the low-affinity agonist site mediating cyclic GMP formation.毒蕈碱反应及在小鼠神经母细胞瘤克隆株（N1E - 115）中的结合。介导环鸟苷酸形成的低亲和力激动剂位点在传代培养时选择性丧失。
Mol Pharmacol. 1984 Sep;26(2):156-63.
3
Histamine H1 receptor-mediated guanosine 3',5'-monophosphate formation by cultured mouse neuroblastoma cells.
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4
Involvement of calcium channels in short-term desensitization of muscarinic receptor-mediated cyclic GMP formation in mouse neuroblastoma cells.钙通道在小鼠神经母细胞瘤细胞毒蕈碱受体介导的环磷酸鸟苷形成的短期脱敏中的作用。
Proc Natl Acad Sci U S A. 1980 Nov;77(11):6897-901. doi: 10.1073/pnas.77.11.6897.
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Voltage-sensitive calcium channels regulate guanosine 3',5'-cyclic monophosphate levels in neuroblastoma cells.电压敏感性钙通道调节神经母细胞瘤细胞中环磷酸鸟苷水平。
Proc Natl Acad Sci U S A. 1978 Dec;75(12):6295-9. doi: 10.1073/pnas.75.12.6295.
6
Regulation of synthesis of guanosine 3':5'-cyclic monophosphate in neuroblastoma cells.神经母细胞瘤细胞中3':5'-环磷酸鸟苷合成的调控
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Muscarinic responses and binding in a murine neuroblastoma clone (N1E-115). Mediation of separate responses by high affinity and low affinity agonist-receptor conformations.小鼠神经母细胞瘤克隆株（N1E-115）中的毒蕈碱反应与结合。高亲和力和低亲和力激动剂-受体构象对不同反应的介导作用。
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8
Phorbol ester-induced inhibition of cyclic GMP formation mediated by muscarinic receptors in murine neuroblastoma cells.
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9
Inhibition by lithium of cyclic GMP formation without inhibition of nitric oxide generation in the mouse neuroblastoma cell (N1E-115).锂对小鼠神经母细胞瘤细胞（N1E - 115）中环鸟苷酸形成的抑制作用，而不抑制一氧化氮的生成。
Neuropsychopharmacology. 1994 Oct;11(2):119-24. doi: 10.1038/npp.1994.41.
10
Effects of cholinergic agents and sodium ions on the levels of guanosine and adenosine 3':5'-cyclic monophosphates in neuroblastoma and neuroblastoma X glioma hybrid cells.
FEBS Lett. 1975 Nov 1;59(1):74-9. doi: 10.1016/0014-5793(75)80344-9.

本文引用的文献

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Neurotransmitter synthesis by neuroblastoma clones (neuroblast differentiation-cell culture-choline acetyltransferase-acetylcholinesterase-tyrosine hydroxylase-axons-dendrites).神经母细胞瘤克隆体的神经递质合成（神经母细胞分化 - 细胞培养 - 胆碱乙酰转移酶 - 乙酰胆碱酯酶 - 酪氨酸羟化酶 - 轴突 - 树突）
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Effects of collagen and of aspirin on the concentration of guanosine 3':5'-cyclic monophosphate in human blood platelets: measurement by a prelabelling technique.胶原蛋白和阿司匹林对人血小板中3':5'-环磷酸鸟苷浓度的影响：采用预标记技术进行测定
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Elevation of cyclic guanosine 3',5'-monophosphate levels in dog thyroid slices caused by acetylcholine and sodium fluoride.乙酰胆碱和氟化钠引起犬甲状腺切片中环磷酸鸟苷水平升高。
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Radioimmunoassay for cyclic nucleotides. I. Preparation of antibodies and iodinated cyclic nucleotides.环核苷酸的放射免疫测定。I. 抗体及碘化环核苷酸的制备。
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8
Separation of cyclic 3',5'-nucleoside monophosphates from other nucleotides on aluminum oxide columns. Application to the assay of adenyl cyclase and guanyl cyclase.在氧化铝柱上从其他核苷酸中分离环3',5'-单磷酸核苷。应用于腺苷酸环化酶和鸟苷酸环化酶的测定。
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A simple, sensitive protein-binding assay for guanosine 3':5'-monophosphate.一种用于3':5'-环磷酸鸟苷的简单、灵敏的蛋白质结合测定法。
Proc Natl Acad Sci U S A. 1971 Apr;68(4):736-9. doi: 10.1073/pnas.68.4.736.
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A radioisotopic method for measuring the formation of adenosine 3',5'-cyclic monophosphate in incubated slices of brain.一种用于测量脑切片孵育过程中3',5'-环磷酸腺苷形成的放射性同位素方法。
J Neurochem. 1969 Dec;16(12):1609-19. doi: 10.1111/j.1471-4159.1969.tb10360.x.

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