Analysis of calcium oscillations in vascular smooth muscle cells

血管平滑肌细胞钙振荡分析

• 批准号: 07457021
• 负责人: IINO Masamitsu
• 金额: $ 4.61万
• 依托单位: The University of Tokyo
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 1995
• 资助国家: 日本
• 起止时间: 1995 至 1996
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07457021/
• 关键词: Vascular smooth muscle; Calcium ion; Inositol trispohophate; Blood pressure regulation; Digital imaginf; Fluorescent Ca^<2+> indicator; Ca^<2+> channel antagonist; 蛍光カルシウム指示薬

Using cooled CCD camera or confocal laser scanning microscope, we carried out digital imaging of intracellular Ca^<2+> concentration of vascular smooth muscle cells within intact arterial tissue. Ca^<2+> indicator loaded arterial tissue was mounted in an experimental chamber on the stage of an inverted microscope and stimulated with field electrical stimulation or by solution exchange. We could observe Ca^<2+> concentration change within individual smooth muscle cells, and confirmed our previous results that the intracellular Ca^<2+> concentration change takes place as Ca^<2+> oscillations and waves when the arterial tissue was stimulated with the sympathetic nerve transmitter, noradrenaline. The current experimental system had a greater dynamic range than that in our previous measurements. We studied the effect of dihydropyridine Ca^<2+> channel antagonists on the Ca^<2+> oscillations. In the presence of nicardipine at a concentration enough to inhibit high K-induced intracellular Ca^<2+> increase in vascular smooth muscle cells the frequency of the noradrenaline-induced Ca^<2+> oscillation was significantly inhibited. This new finding suggest that Ca^<2+> channel antagonist can inhibit Ca^<2+> store-mediated Ca^<2+> signalling. We are now expanding our work and preparing for simultaneous measurement of intracellular Ca^<2+> concentrations of vascular smooth muscle cells and endothelial cells to study the effect of endothelium derived relaxing factor on the Ca^<2+> signalling in vascular smooth muscle cells.

使用冷却的CCD摄像头或共聚焦激光扫描显微镜，我们对完整动脉组织内血管平滑肌细胞的细胞内CA^<2+>进行了数字成像。 Ca^<2+>指示剂负载的动脉组织被安装在倒置显微镜阶段的实验室中，并用场电刺激或溶液交换刺激。我们可以观察到单个平滑肌细胞内的Ca^<2+>浓度变化，并证实了我们先前的结果，即当动脉组织与交感神经透射蛋白促进动脉组织刺激时，细胞内Ca^<2+>浓度变化发生在Ca^<2+>振荡和波浪中。当前的实验系统的动态范围比我们先前测量的动态范围更大。我们研究了二氢吡啶Ca^<2+>通道拮抗剂对CA^<2+>振荡的影响。在足够浓度以抑制高k诱导的细胞内Ca^<2+>血管平滑肌细胞增加的症状的存在下，去甲肾上腺素诱导的Ca^<2+>振荡的频率显着抑制。这一新发现表明Ca^<2+>通道拮抗剂可以抑制Ca^<2+>商店介导的Ca^<2+>信号传导。现在，我们正在扩大工作，并准备同时测量血管平滑肌细胞和内皮细胞的细胞内CA^<2+>浓度，以研究内皮衍生的放松因子对血管平滑肌细胞中CA^<2+>信号传导的影响。

项目成果

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