Development of the System for Measurement of Intramyocardial Ion Dynamics in Isolated, Perfused Hearts

离体灌注心脏心肌内离子动力学测量系统的开发

• 批准号: 07558123
• 负责人: KUSUOKA Hideo
• 金额: $ 7.49万
• 依托单位: Osaka University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (A)
• 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-07558123/
• 关键词: Myocardium; Intracellular Ion Concentration; Perfused Heart; Fluorescent Dye; 灌流心

It is important to measure intramyocardial ion dynamics for the investigation of physiology and pathophysiology of hearts. The measurements of intracellular ion concentration, the method using fluorescent dyes has advantages including high temporal resolution. However, it has been difficult to apply fluorescent dyes into isolated, perfused hearts. To load dyes to perfused hearts, we developed the system using the iontophoresis technique. In this study, we completed the following items ;1.We optimized the method to produce microelectrodes for iontophoresis of dyes.2.We determined the optimal conditions for the current and the optinal positioning of the electrode.3.We developed the method to stop or attenuate the beating of the hearts. First, the calcium concentration in the perfusate was decreased, and then, potasium concentration was increased or BDM was added to the perfusate.4.In our method, microelectrode was induced above the heart, and iontophoresis was performed from the top of the stage of microscope. In contrast, the ultraviolet beam was introduced to the heart from the bottom of the microscope. Thus, it was necessary to change the orientation of the heart between the procedures, and caused the disorientation. To avoid this, we developed the method to keep the orientation of hearts.Our study indicates the feasibility of the iontophoresis in isolated, perfused hearts, and we accomplished some basic part of the techniques required for this approach. Nevertheless, there are still many remaining problems to make this technique more practical.In addition, we also evaluated the different effects of proteins on Ca^<2+> ion measurements using fluorescent dye or NMR technique.

心肌内离子动力学的测定对心脏生理和病理生理的研究具有重要意义。在细胞内离子浓度的测量中，使用荧光染料的方法具有包括高时间分辨率的优点。然而，将荧光染料应用于离体灌注的心脏是困难的。为了将染料加载到灌注的心脏中，我们开发了使用离子电渗技术的系统。本研究主要完成了以下几个方面的工作：1.优化了染料离子导入微电极的制备方法; 2.确定了最佳的电流条件和电极的最佳位置; 3.建立了使心脏停止或减弱跳动的方法。先降低灌流液中钙离子浓度，再升高钾离子浓度或加入BDM。4.在心脏上方诱导微电极，从显微镜载物台上方进行离子导入。相比之下，紫外线光束从显微镜的底部引入心脏。因此，有必要在手术之间改变心脏的方向，并导致定向障碍。为了避免这种情况的发生，我们开发了保持心脏方向的方法，我们的研究表明了离子导入在离体灌流心脏中的可行性，并完成了这种方法所需的一些基本技术。此外，我们还研究了蛋白质对荧光染料法和核磁共振法测定Ca^2+的影响。

项目成果

Shinichi Matsuda: "The effects of proteins on Ca^<2+> measurement : different effects on fluorescent and NMR methods" Cell Calcium. 20.5. 425-430 (1996)

Shinichi Matsuda：“蛋白质对 Ca^2 测量的影响：对荧光和 NMR 方法的不同影响”细胞钙。

Shinichi Matsuda: "The effects of proteins on Ca^<2+> measurement : different effects on in fluorescent and NMR methods" Cell Calcium. 20.5. 425-430 (1996)

Shinichi Matsuda：“蛋白质对 Ca^2 测量的影响：对荧光和 NMR 方法的不同影响”细胞钙。

Shinichi Matsuda: "The effects of proteins on 〔Ca^<2+>〕 measurement : different effects on fluorescent and NMR methods" Cell Calcium. 20.5. 425-430 (1996)

Shinichi Matsuda：“蛋白质对[Ca^<2+>]测量的影响：对荧光和NMR方法的不同影响”细胞钙20.5（1996）。