Development of a measurement system of autonomic nervous activity in genetic pathological-model mice

遗传病理模型小鼠自主神经活动测量系统的研制

• 批准号: 10557009
• 负责人: KANNAN Hiroshi
• 金额: $ 2.94万
• 依托单位: Miyazaki Medical College
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 1998
• 资助国家: 日本
• 起止时间: 1998 至 2001
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-10557009/
• 关键词: autonomic nervous activity; cardiovascular system; awake animals; renal excretory function; hypothalamic single unit activity; unit recording electrode; micro-movable manipulator; multiple data processing; 遺伝性多飲マウス; アンジオテンシンII; 循環系応答; 飲水行動; 動脈

Techniques to create transgenic organisms or animals with targeted mutations ('knockout' mutants) have become increasingly important tools in neuroscience over the past few years. As always, new techniques, besides providing new tools to investigate problems or to test hypothesis, also give rise to unforeseen difficulties which take time to overcome. So far, most genetic animals constructed have been mice, whose brains are quite hard to manipulate (e.g., stimulation of discrete areas and unit recordings are difficult to achieve with mice in the freely-moving condition). Therefore, we devised the following procedures to perform physiological studies in mice.(1) Construction of a metabolic cage for small animals such as mice that is regulated by means of a personal computer and allows the experimenter to record cardiovascular parameters without disturbing the animal's movement or twisting the recording lead wires and vascular catheters. For studies involving the central nervous system, i … More t is often desirable to record, stimulate, and/or deliver drugs to discrete regions of the brain. Although this can be carried out somewhat routinely in anesthetized preparations, it has long been recognized that anesthesia may confound the interpretation of studies of the central regulation of cardiovascular function and body fluid balance. Therefore, it was necessary to develop a procedure for carrying out similar experiments in animals that are awake. The floor of the plastic cage was upturned to release the twist of the cable leads and catheters as the animal moved ; this was performed by signals detected with an inertia sensor.(2) A system for simultaneously displaying multiple parameters on a monitor screen with variable sweep speed was constructed to elucidate the long-term trend of signals whereby the amplitude of the action potentials and the noise levels provide cue information about the relative location of the recording electrodes and neurons under study. This system is useful to evaluate the advancement of the recording electrodes. In addition, computer programs for multi-variable analysis and three-dimensional illustrations were constructed.(3) Recording methods for single-unit activity in freely-moving animals have been improved as follows : (a) First, we improved the recording electrodes, which are made of stainless micro wires coated with Formvar. We found that electrodes with high impedance are suitable to record single-unit activity with a high signal-to noise ratio efficiently. Therefore, stainless micro wires were insulated several times with a thin Epoxy solution, (b) Second, a miniature skull-mounted microdrive that permits discrete dorsoventral movement of the fine-wire recording electrodes was constructed to advance the fine-wire electrode as well as to isolate single-unit activities, (c) Third, a head-mounted voltage follower was employed to condition and stabilize the neuronal signals prior to transmission or for further signal processing. These methods were followed in order to integrate these three elements into a working electrophysiological recording procedure in freely-moving small animals. Less

在过去的几年里，创造具有靶向突变（“敲除”突变体）的转基因生物或动物的技术已经成为神经科学中越来越重要的工具。与以往一样，新技术除了提供新的工具来研究问题或检验假设外，也会引起无法预料的困难，需要时间来克服。到目前为止，大多数构建的遗传动物都是老鼠，它们的大脑很难操纵（例如，离散区域的刺激和单位记录难以用处于自由移动条件下的小鼠实现）。因此，我们设计了以下程序来在小鼠中进行生理学研究。(1)为小鼠等小动物构建代谢笼，通过个人计算机进行调节，并允许实验者在不干扰动物运动或扭曲记录引线和血管导管的情况下记录心血管参数。对于涉及中枢神经系统的研究，i ...更多信息 通常需要记录、刺激和/或递送药物到脑的离散区域。虽然这可以在麻醉制剂中进行，但人们早就认识到麻醉可能会混淆对心血管功能和体液平衡的中枢调节研究的解释。因此，有必要开发一种在清醒的动物中进行类似实验的程序。当动物移动时，将塑料笼的地板向上翻，以释放电缆导线和导管的扭曲;这是通过惯性传感器检测到的信号来执行的。(2)一个系统，用于同时显示多个参数在监视器屏幕上与可变的扫描速度，以阐明信号的长期趋势，从而动作电位的幅度和噪声水平提供线索信息的相对位置的记录电极和神经元的研究。该系统可用于评估记录电极的进展。此外，还编制了多变量分析和三维图形的计算机程序。(3)自由运动动物的单单位活动的记录方法已经改进如下：（a）首先，我们改进了记录电极，其由涂覆有Formvar的不锈钢微丝制成。我们发现，具有高阻抗的电极适合于以高信噪比有效地记录单个单位活动。因此，用薄环氧树脂溶液将不锈钢微丝绝缘若干次。（B）第二，构造允许细线记录电极的离散背腹移动的微型颅骨安装微驱动器，以推进细线电极以及隔离单个单元活动。（c）第三，使用头戴式电压跟随器在传输之前调节和稳定神经元信号或用于进一步的信号处理。遵循这些方法，以便将这三个元素整合到自由移动的小动物的工作电生理记录程序中。少

项目成果

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