Daniels A J, Lazarowski E R, Matthews J E, Lapetina E G
Medicinal Biochemistry Division, Wellcome Research Laboratories, Research Triangle Park, N.C. 27709.
Biochem Biophys Res Commun. 1989 Dec 29;165(3):1138-44. doi: 10.1016/0006-291x(89)92721-6.
The intracellular concentration of free Ca2+ was monitored by measuring the fluorescence of fura-2 loaded Human Erythroleukemia Cells. Neuropeptide Y (NPY) increased intracellular Ca2+ in a dose-dependent manner and the 50% effective concentration was 2 nM. Chelation of extracellular Ca2+ by EGTA did not reduce the NPY-mediated increase in cytoplasmic Ca2+, indicating that the increase in fluorescence was due to the release of intracellular Ca2+. A second dose of NPY, after intracellular Ca2+ had returned to basal levels, failed to elicit a response, indicating that the NPY receptor had undergone desensitization. In similar experiments, NPY increased the formation of inositol phosphates, suggesting that the mobilization of Ca2+ from intracellular stores in HEL cells was secondary to the generation of inositol phosphates and stimulation of phospholipase C.
通过测量负载fura-2的人红白血病细胞的荧光来监测细胞内游离Ca2+的浓度。神经肽Y(NPY)以剂量依赖性方式增加细胞内Ca2+,50%有效浓度为2 nM。EGTA对细胞外Ca2+的螯合作用并未降低NPY介导的细胞质Ca2+增加,表明荧光增加是由于细胞内Ca2+的释放。在细胞内Ca2+恢复到基础水平后,再次给予NPY未能引发反应,表明NPY受体已发生脱敏。在类似实验中,NPY增加了肌醇磷酸的形成,提示HEL细胞中细胞内储存的Ca2+的动员继发于肌醇磷酸的生成和磷脂酶C的刺激。