Integrated analysis of three-dimensional molecular dynamics of actin and myosin, and macro-dynamics of cardiac muscle and left ventricle

肌动蛋白、肌球蛋白三维分子动力学、心肌、左心室宏观动力学综合分析

• 批准号: 15300175
• 负责人: TSUJIOKA Katsuhiko
• 金额: $ 10.37万
• 依托单位: Kawasaki Medical School
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2003
• 资助国家: 日本
• 起止时间: 2003 至 2005
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-15300175/
• 关键词: synchrotron radiation x-ray; cardiac muscle; actin; myosin head; crossbridge; x-ray diffraction; laser diffraction; srcomere; レーザ回折

The purpose of this project is to analyze the molecular interaction between actin and myosin in isolated cardiac muscle and whole heart with time- resolved way in one cardiac cycle.We assembled the short angle diffraction apparatus with x-ray energy 12.4keV and wave length 1 A at BL45XU beam line in Spring-8 (Super Photon Ring 8GeV). We measured x-ray diffraction pattern by using x-ray image intensifier and cooled CCD. For the measurement of diffraction pattern from the left ventricle we used BL40XU, which has higher energy than BL45XU. In this case the camera length was 3m and x-ray energy was 16keV. We controlled the x-ray energy to the suitable strength with changing the width of front-end slit. We measured the sarcomere length by laser diffraction pattern and compared it with lateral diffraction pattern by x-ray, and thus analyze the three-dimensional dynamics between molecular and cellular level.We got the equatorial diffraction from isolated and perfused papillary muscle from rig … More ht ventricle of rat. From the ratio between (1,0)/(1,1) diffraction the mass transfer from the myosin molecule to actin molecule, which is the index of crossbridge formation, was measured. We analyzed the change of molecular interaction between actin and myosin with the change of sarcomere length and afterload in isometric and isotonic contraction. In addition we analyzed the x-ray diffraction pattern from left ventricular wall in isolated Langendorff-perfusion heart.As the mass transfer from myosin molecular to actin increased with cardiac muscle length in isometric contraction, we concluded the amount of crossbridge formation is the molecular basis of the Starling mechanism of the heart. In isotonic contraction the developed force also increased with the amount of crossbridge formation. However, the amount of mass-transfer from the myosin molecular to actin in small afterloaded contraction was interestingly smaller than that at the same developed force in isometric contraction. The developed pressure in isovolumically contracting left ventricle was in accordance with the mass-transfer, indicating the molecular basis of the Staring mechanism of the heart in whole heart. The diffraction pattern from the left ventricular wall changed with regular manner depending on x-ray irradiation portion at subepicardium, mid wall and subendocardium. We concluded that the change of the three-dimensional molecular structure at the different portion of the left ventricular free wall could be estimated in time resolved manner in isovolumically contracting isolated and perfused heart. Less

本课题的目的是在Spring-8（Super Photon Ring 8 GeV）的BL-45 XU束线上组装了能量为12.4keV、波长为1A的短角X射线衍射装置，在一个心动周期内，用时间分辨的方法分析离体心肌和整个心脏中肌动蛋白和肌球蛋白之间的分子相互作用。我们用X射线像增强器和冷却CCD测量了X射线衍射图。对于来自左心室的衍射图案的测量，我们使用BL 40 XU，其具有比BL 45 XU更高的能量。在这种情况下，相机长度为3米，X射线能量为16 keV。通过改变前端狭缝的宽度来控制X射线能量到合适的强度。我们用激光衍射图测量了乳头肌的肌节长度，并与X射线横向衍射图进行了比较，从而分析了分子水平和细胞水平的三维动力学。 ...更多信息 HT大鼠心室。从（1，0）/（1，1）衍射之间的比率，测量从肌球蛋白分子到肌动蛋白分子的质量传递，这是横桥形成的指数。我们分析了在等长收缩和等张收缩中，随着肌节长度和后负荷的变化，肌动蛋白和肌球蛋白分子间相互作用的变化。此外，我们还分析了Langendorff灌流心脏左室壁的X-射线衍射图谱，发现在等长收缩过程中，随着心肌长度的增加，肌球蛋白分子向肌动蛋白的质量传递增加，因此，心肌心肌桥的形成是心脏Starling机制的分子基础。在等张收缩中，所产生的力也随着横桥形成量的增加而增加。然而，有趣的是，在小的后负荷收缩中，从肌球蛋白分子到肌动蛋白的质量转移量小于在等长收缩中相同的力。等容收缩时左心室内的压力变化与质量传递过程一致，表明心脏在全心脏范围内存在凝视机制的分子基础。心外膜下、中壁和内膜下X线照射部位不同，左室壁衍射花样呈规律性变化。我们的结论是，在等容收缩离体心脏和灌流的左心室游离壁的不同部分的三维分子结构的变化可以估计在时间分辨的方式。少

项目成果

An X-ray diffraction study on contraction of rat papillary muscle with different afterloads.

不同后负荷大鼠乳头肌收缩的 X 射线衍射研究。

• 发表时间: 2003
• 期刊: Adv Exp Med Biol 538
• 作者: Saito K;et al.;Okuyama H
• 通讯作者: Okuyama H

Analysis of crossbridge dynamics of left ventricular free wall in beating heart.

心脏跳动时左心室游离壁的横桥动力学分析。

• 发表时间: 2005
• 期刊: Spring-8 User Experiment Report No.14(2004B)
• 作者: Fumihiko Kajiya;Naoto Yagi;Juichiro Shimizu;Satoshi Mohri;Kazufumi Nakamura;Katsushi Hashimoto;Taro Morimoto;Hiroshi Okuyama;Hiroko Toyota;Katsuhiko Tsujioka
• 通讯作者: Katsuhiko Tsujioka

Transmurally difference in crossbridge dynamics and sarcemere shortening during relaxation of isovolumic beat

等容搏动松弛过程中跨桥动力学和肌节缩短的跨壁差异

• 发表时间: 2005
• 期刊: Circulation Journal 69:Suppl I
• 作者: Juichiro S;et al.
• 通讯作者: et al.

Direct observation and quantitative analysis of spatiotemporal dynamics of individual living monocytes during transendothelial migration

活单核细胞跨内皮迁移过程中时空动态的直接观察和定量分析

• 发表时间: 2004
• 期刊: Atherosclerosis 177・1
• 作者: Ken Hashimoto

Mochizuki S, Miyasaka T, Goto M, Ogasawara Y, Yada T, Akiyama M, Neishi Y, Toyoda T, Tomita J, Koyama Y, Tsujioka K, Kajiya F, Akasaka T, Yoshida K: "Measurement of acetylcholine-induced endothelium-derived nitric oxide in aorta using a newly developed ca

Mochizuki S、Miyasaka T、Goto M、Ogasawara Y、Yada T、Akiyama M、Neishi Y、Toyoda T、Tomita J、Koyama Y、Tsujioka K、Kajiya F、Akasaka T、Yoshida K：“乙酰胆碱诱导的内皮细胞的测量-