THE DEVELOPMENT OF LASER-FLUOROMETRIC MICROSCOPE FOR MEASUREMENT OF INTRACELLULAR CALCIUM IONS SINGLE LIVING CELLS.

用于测量单个活细胞内钙离子的激光荧光显微镜的开发。

• 批准号: 63870038
• 负责人: YAZAKI Yoshio
• 金额: $ 15.36万
• 依托单位: TOKYO UNIVERSITY, FACULTY OF MEDICINE.
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Developmental Scientific Research
• 财政年份: 1988
• 资助国家: 日本
• 起止时间: 1988 至 1989
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-63870038/
• 关键词: INTRACELLULAR CALCIUM IONS; HYPERTENSION; LASER MICROSCOPE; VASCULAR SMOOTH MUSCLE CELLS; INDO-1; SPONTANEOUSLY HYPERTENSIVE RATS; ABNORMAL CALCIUM HANDLING; INTRACELLULAR CALCIUM ION ELEVATING FACTOR; インドー1(INDC-1); レーザー螢光顕微鏡; Indoー1; 血管平滑筋

We have developed an accurate and sensitive system for the measurement of cytoplasmic free calcium concentrations ([Ca^<++>]_i) of a single vascular smooth muscle cell (VSMC) by using UV-laser and Indo-1. [Ca^<++>]_i in spontaneously hypertensive rat (SHR) was elevated and maintained after the 6th passage culture. And moreover, marked elevation of [Ca^<++>]_i after stimulation with caffeine and angiotensin-II occurred in VSMC in some populations of 4-week-old SHR even when blood pressure and basal [Ca^<++>]_i had not yet been elevated. In 8-week-old rats, the basal [Ca^<++>]_i from SHR was elevated compared with Wistar Kyoto rats. We have attempted to characterize yet unspecified circulating factor(s) from SHR serum that elevates [Ca^<++>]_i in VSMC. Our results showed that 1) the maintenance of high [Ca^<++>]_i levels in VSMC in SHR is genetically regulated and it is one of the mechanisms in SHR hypertension, 2) abnormal calcium regulation in VSMC from SHR is expressed even prior to overt hypertension, and 3) the [Ca^<++>]_i elevating factor(s) exists in 8W-SHR serum, but not in 4W-SHR, 4W- or 8W- WKY serum.

我们建立了一个用紫外激光和Indo-1测定单个血管平滑肌细胞（VSMC）胞浆游离钙浓度（[Ca^++]_i）的系统。[Ca自发性高血压大鼠（SHR）心肌细胞的[M]_i在第6代时升高并维持不变。此外，在血压和基础[Ca ^<++>] i尚未升高的情况下，咖啡因和血管紧张素-II刺激4周龄SHR的某些群体的VSMC，可使[Ca ^<++>] i显著升高。在8周龄大鼠中，SHR的基础[Ca^<++>]_i高于Wistar京都大鼠。我们试图从SHR血清中鉴定升高VSMC [Ca^<++>]_i的未指明的循环因子。结果表明：（1）SHR血管平滑肌细胞内高[Ca^++] i水平的维持受遗传调节，是SHR高血压的机制之一;（2）SHR血管平滑肌细胞内钙调节异常早于高血压发生;（3）8 W-SHR血清中存在[Ca^++] i升高因子，而4 W-SHR血清中不存在[Ca^++] i升高因子。4 W-或8 W- WKY血清。

项目成果

Sugiyama T.Yoshizumi M.: "Abnormal calcium handling in vascular smooth muscle cells in spontaneously hypertensive rats." J.Hypertens.

Sugiyama T.Yoshizumi M.：“自发性高血压大鼠血管平滑肌细胞中的钙处理异常。”

Komuro I.Kurihara H.他: "Endothelin stimulate c-fos and c-myc expression and proliferation of vascular smooth muscle cells." FEBS Lett.238. 249-252 (1988)

Komuro I. Kurihara H. 等人：“内皮素刺激血管平滑肌细胞的 c-fos 和 c-myc 表达和增殖。” FEBS Lett.238 (1988)。

Komuro I, Kurihara H, Sugiyama T, Yoshizumi M, Takaku F, Yazaki Y.: "Endothelin stimulate c-fos and c-myc expression and proliferation of vascular smooth muscle cells." FEBS Lett.238. 249-252 (1988)

Komuro I、Kurihara H、Sugiyama T、Yoshizumi M、Takaku F、Yazaki Y.：“内皮素刺激 c-fos 和 c-myc 表达以及血管平滑肌细胞的增殖。”

Yanagisawa M.Kurihara H.他: "A novel potent vasoconstrictor peptide produced by vascular endothelial cells." Nature.332. 411-415 (1988)

Yanagisawa M. Kurihara H. 等人：“一种由血管内皮细胞产生的新型有效血管收缩肽。”Nature.332-415（1988）。

Yoshizumi M.Kurihara H.: "Hemodynamic shear stress stimulates endothelin production by cultured endothelial cells." Biochem Biophys Res Commun.161. 859-864 (1989)

Yoshizumi M.Kurihara H.：“血流动力学剪切应力刺激培养的内皮细胞产生内皮素。”