Micromanipulators and Microscope Isolation Stage

显微操作器和显微镜隔离台

• 批准号: 440393-2013
• 负责人: Buck, Leslie
• 金额: $ 1.64万
• 依托单位: University of Toronto
• 依托单位国家: 加拿大
• 项目类别: Research Tools and Instruments - Category 1 (<$150,000)
• 财政年份: 2012
• 资助国家: 加拿大
• 起止时间: 2012-01-01 至 2013-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=506189
• 关键词: Micromanipulators; Microscope; Isolation; Stage

The Buck lab is investigating cellular mechanisms that permit some species, such as freshwater turtles, goldfish and snails to survive without oxygen (anoxia) for months during winter. We use both a liver and brain cell model to study the mechanisms of anoxia tolerance. Key to surviving anoxia is the shutting off of energy utilizing cellular activities, such as the synthesis of proteins and maintaining ion gradients across cell membranes. We have previously shown that protein synthesis decreases by 90%, the pump that maintains ion gradients decreases by 75%, and ion channel activity (pathways for ions to cross membranes) decreases (in one case increases). The ability to reduce the rate of these energy consuming reactions is something that mammals (humans) lack. However, basic biochemical pathways are common to almost all species, certainly amongst reptiles, fish, birds and mammals. Our goal is to determine the natural cellular pathways responsible for shutting off energy consuming processes in anoxia-tolerant species. We have already identified several possible pathways and discovered that brain excitatory signaling involving glutamate decreases, while inhibitory activity involving GABA is increases during anoxia. These results are remarkably similar to the way in which volatile anesthetics reduce brain activity in humans and we may have uncovered a natural anesthetic mechanism. Brain is an electrically excitable tissue and we are also investigating anoxic regulation of ion channels in liver, a non-excitable tissue. We will continue to explore the electrophysiological properties of brain and liver tissue to better understand which ion channels are regulated in a way that reduces energetic demands and permits anoxic survival. The equipment requested in this proposal is the basis of a fundamental electrophysiological technique in the Buck Lab to explore mechanisms of anoxia tolerance; however, it is a dual electrode system that requires two motorized micromanipulators for the fine scale positioning of electrodes and a sturdy microscope isolation stage to attach the manipulators. The new equipment will maintain and increase my research productivity and support the research and training of an increased number of students and postdoctoral fellows.

巴克实验室正在研究细胞机制，使一些物种，如淡水龟，金鱼和蜗牛在冬季无氧（缺氧）生存数月。我们使用肝脏和脑细胞模型来研究缺氧耐受的机制。缺氧存活的关键是关闭利用细胞活动的能量，例如蛋白质的合成和维持细胞膜上的离子梯度。我们之前已经表明，蛋白质合成减少了90%，维持离子梯度的泵减少了75%，离子通道活性（离子穿过膜的途径）降低（在一种情况下增加）。降低这些能量消耗反应速率的能力是哺乳动物（人类）所缺乏的。然而，基本的生物化学途径几乎对所有物种都是共同的，当然包括爬行动物、鱼类、鸟类和哺乳动物。我们的目标是确定在耐缺氧物种中负责关闭能量消耗过程的天然细胞途径。我们已经确定了几种可能的途径，并发现在缺氧期间，涉及谷氨酸的大脑兴奋性信号减少，而涉及GABA的抑制性活动增加。这些结果与挥发性麻醉剂减少人类大脑活动的方式非常相似，我们可能已经发现了一种天然的麻醉机制。大脑是一个电兴奋的组织，我们也正在研究缺氧调节的离子通道在肝脏，一个非兴奋组织。我们将继续探索脑和肝组织的电生理特性，以更好地了解哪些离子通道以减少能量需求并允许缺氧存活的方式进行调节。本提案中要求的设备是巴克实验室探索缺氧耐受机制的基本电生理技术的基础;然而，它是一个双电极系统，需要两个电动显微操作器用于电极的精细尺度定位和一个坚固的显微镜隔离台来连接操作器。 新设备将保持和提高我的研究生产力，并支持更多的学生和博士后研究员的研究和培训。