DEVELOPMENT OF A NEW EXPERIMENTAL APPARATUS FOR STUDYING OF MECHANICAL ACTIVITY OF ISOLATED ARTERIOLES.

开发用于研究离体小动脉机械活动的新实验装置。

• 批准号: 07557056
• 负责人: OHHASHI Toshio
• 金额: $ 1.86万
• 依托单位: Shinshu University School of Medicine
• 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-07557056/
• 关键词: pressurized arteriole; mechanical activity; outer diameter; inner diameter; smooth muscle cells; endothelial cells; vasoactive substances; apparatus for studying mechanical activity of isolated arterioles; 血管内皮細胞; 内皮細胞; 特殊臓器槽; 生体顕微鏡; 微小ガラスカニュー

[1] DEVELOPMENT OF A NEW EXPERIMENTAL APPARATUS FOR STUDYING OF MECHANICAL ACTIVITY OF ISOLATED ARTERIOLESPreparation and dissectionExperimental materials, i. e., brain, spinal cord, or chick pouch, were rapidly removed and placed in a cooled dissection chamber filled with MOPS-buffered physiological salt solution containing 1% BSA.The chamber is placed on a recessed Plexiglas well, and chilled fluid is circulated through the well jacket from a cooling thermopump. The isolated arteriole was carefully transferred from the dissection chamber to a temperature-controlled cannulation chamber mounted on a stage of an inverted microscope.CannulationThe isolated arteriole was cannulated at both ends by using a system of concentric glass pipettes mounted on a micromanipulator which was equipped on the stage of the inserted microscope. By applying vacuum to the lumen of the holding pipette, one end of the arteriole was gently pulled into the pipette. The perfusion pipette was inserted into the l … More umen of the arteriole. Then the other end of the arteriole was cannulated in the same manner.InstrumentationAfter the cannulation, both micropipettes were connected to water manometers used to adjust intraluminal pressure via independent reservoirs. Another fluid reservoir pressurized by the manometer could be inserted into the fluid path between the arteriole and the upstream reservoir by menas of a three-way liquid switch.The image of the cannulated arteriole was recorded using a video camera and displayd on a television monitor. Arteriolar luminal diameters were measured manually using a video caliper incorporated with MacLab Chart data acquisition system.[2] PHYSIOLOGICAL APPLICATIONWe investigated the responses of isolated rabbit spinal arterioles to flow with special reference to endogenous nitric oxide (NO) and prostaglandins (PGs). The arterioles (-100mum in diameter) were cannulated at both ends with microglass pipettes, and perfused at constant pressure (-60mmHg). The arterioles developed spontaneous tone at 37ﾟC by -26% of their maximum dilation. Treatment with indomethacin (10mum) produced a significant decrease in the arteriolar diameter, whereas treatment with a nitric oxide production inhibitor (L-NAME) produced no significant change. In the presence of 10muM indomethacin, however, 30muM L-NAME caused a marked decrease in the arteriolar diameter, which was completely reversed with additional administration of 1mM L-arginine. These findings suggest that both NO and vasodilative PGs released from the endothelial cells may play important roles in the flow-mediated vasodilation in the rabbit spinal arterioles. Less

[1] 开发一种用于研究离体小动脉机械活动的新型实验装置准备和解剖实验材料，i．例如，大脑、脊髓或雏鸡育儿袋，被快速取出并放置在充满含有 1% BSA 的 MOPS 缓冲生理盐溶液的冷却解剖室中。该室放置在凹进的有机玻璃孔上，冷冻流体从冷却热泵通过孔夹套循环。将分离的小动脉小心地从解剖室转移到安装在倒置显微镜载物台上的温控插管室。插管使用安装在显微操作器上的同心玻璃移液管系统在两端对分离的小动脉进行插管，显微操作器配备在插入显微镜的载物台上。通过向固定移液管的内腔施加真空，将小动脉的一端轻轻拉入移液管中。将灌注移液管插入小动脉腔内。然后以相同的方式对小动脉的另一端进行插管。插管后，将两个微量移液器连接到水压计，用于通过独立的储液器调节腔内压力。另一个由压力计加压的流体储存器可以通过三路液体开关插入到小动脉和上游储存器之间的流体路径中。使用摄像机记录插管小动脉的图像并显示在电视监视器上。使用与 MacLab Chart 数据采集系统结合的视频卡尺手动测量小动脉管腔直径。 [2]生理学应用我们研究了离体兔脊髓小动脉对血流的反应，特别是内源性一氧化氮（NO）和前列腺素（PG）。用微型玻璃吸管在小动脉（直径-100μm）的两端插管，并在恒压（-60mmHg）下灌注。小动脉在 37°C 时产生自发张力，其最大扩张度为 -26%。用吲哚美辛 (10mum) 治疗可使小动脉直径显着减小，而用一氧化氮生成抑制剂 (L-NAME) 治疗则没有产生显着变化。然而，在 10μM 吲哚美辛存在的情况下，30μM L-NAME 会导致小动脉直径显着减小，而另外施用 1mM L-精氨酸则完全逆转了这种情况。这些发现表明，内皮细胞释放的 NO 和血管舒张 PG 可能在兔脊髓小动脉的血流介导的血管舒张中发挥重要作用。较少的

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