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TO DEVELOP A NEW SYSTEM TO CONTIMUOUSLY MONITOR THE FUNCTIONS AT THE CELLULAR AND MOLECULAR LEVELS OF THE VASCULAR ENDOTHELIAL AND SMOOTH MUSCLE CELLS IN VIVO.

开发一种新系统，在体内连续监测血管内皮细胞和平滑肌细胞的细胞和分子水平的功能。

基本信息

批准号：
06557045
负责人：
KANAIDE Hideo
金额：
$ 7.87万
依托单位：
KYUSHU UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (A)
财政年份：
1994
资助国家：
日本
起止时间：
1994 至 1996
项目状态：
已结题

项目摘要

1. THE DEVELOPMENT OF A NEW SYSTEM FOR MONITORING THE FUNCTIONS OF THE VASCULAR ENDOTHELIAL AND SMOOTH MUSCLE CELLS : The system we have developed has following characteristics ; (1) Using a grating and a chopper mirror driven by a stepping motor, it became possible to alternately (10 Hz) illuminate the tissue sample with 340 and 380 nm excitation lights for the measurement of intracellular calcium concentration, [Ca] i. (2) the chopper system is controlled through a personal computer by investigator designed software. (3) Fluorescence images are obtained using a chopper mirror and a SIT camera. A display flame is 640x400 dots and a 8 bit flame. (4) Fluorescence ratio (F340/F380) images can be obtained at 5 Hz. (5) It is possible to simultaneously record changes in the [Ca] i levels at 30 points in an image on the TV monitor. (6) Low cost for making the system.2. NEW FINDINGS OBTAINED USING THIS SYSTEM,FRONT-SURFACE FLUORIMETRY AND TECHNIQUES IN MOLECULAR BIOLOGY : (1) Using the newly developed technique, the simultaneous fluorimetry of fura-2 and diaminonaphthalene, we found the [Ca] i-NO production relationships in the endothelial cells in situ. (2) Using front-sueface fluorimetry, reverse transcription polymerase chain reaction, and the renal and coronary arteries, we found that both adrenomedullin and endothelin might act as autocrine/paracrione signal transmitters in these tissues.3. HUMANA PRESS (NJ : U.S.A) asked us to write a chapter "Measurement of [Ca] i in muscle strips using front-surface fluorimetry" of a book on "Calcium signaling protocols (edited by DG Lambert)", a part of the Methods in Molecular Biology series. It is in press.

1.血管内皮细胞和平滑肌细胞功能监测系统的研制：我们研制的系统具有以下特点：（1）使用由步进电机驱动的光栅和斩波镜，可以用340和380 nm激发光交替（10 Hz）照射组织样品，以测量细胞内钙浓度，[Ca] i. (2)斩波器系统通过个人计算机由研究者设计的软件控制。(3)使用斩波镜和SIT相机获得荧光图像。显示火焰是640 x400点和8位火焰。(4)荧光比率（F340/F380）图像可以在5 Hz下获得。(5)可以在电视监视器的图像中同时记录30个点的[Ca] i水平变化。(6)系统制作成本低.利用该系统、前表面荧光法和分子生物学技术获得的新发现：（1）利用新发展的荧光法，即Fura-2和二氨基萘同时荧光法，我们在原位发现了内皮细胞[Ca] i-NO的产生关系。(2)利用荧光光谱法、逆转录聚合酶链反应以及肾动脉和冠状动脉，我们发现肾上腺髓质素和内皮素在这些组织中可能都是自分泌/旁分泌信号传递者. HUMANA PRESS（NJ：U.S.A）要求我们撰写一本关于“钙信号协议（由DG Lambert编辑）”的书的章节“使用前表面荧光法测量肌条中的[Ca] i”，这是分子生物学方法系列的一部分。它正在印刷中。