Developing a zebrafish model of neurovascular coupling (NVC)

开发神经血管耦合 (NVC) 斑马鱼模型

• 批准号: NC/P001173/1
• 负责人: Clare Howarth
• 金额: $ 45.02万
• 依托单位: University of Sheffield
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2016
• 资助国家: 英国
• 起止时间: 2016 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=NC%2FP001173%2F1
• 关键词: Developing; zebrafish; model; neurovascular; coupling

The blood supply to the brain is tightly controlled by a process called "neurovascular coupling". This increases blood supply to areas of the brain which are active, although how this is achieved is not completely understood. Neurovascular coupling is defective in conditions such as stroke and dementia, and understanding this process might ultimately lead to strategies to improve these diseases.Because neurovascular coupling can currently only be studied in a living brain with an intact blood supply, its study requires the use of animals, commonly mice and rats. These experiments require surgery to expose the brain, and imaging of the brain in animals that are either awake and restrained, or anaesthetised, which inevitably induces distress and suffering. We therefore wish to develop a model that can provide scientific insight into neurovascular coupling but reduces the number of invasive animal experiments.The zebrafish has two key advantages for the study of neurovascular coupling. Firstly, because it is transparent and we can generate "transgenic" animals in which certain cells fluoresce different colours, we can image the brain without any instrumentation. Secondly, we can stop the heart without the animal dying, as it obtains sufficient oxygen from the water surrounding it. The tiny zebrafish embryo is not considered to be a free living animal until it is older than 5d, and using these is not considered to be an animal experiment by government legislation (called non-protected).We will exploit these advantages to develop a model of neurovascular coupling in non-protected zebrafish embryos. We have generated genetically modified zebrafish in which either the blood vessels or the nerves fluoresce brightly when levels of calcium in these cells increase. Because calcium levels increase in cells that are "activated", these transgenic lines allow us to image which parts of the brain are activated in response to a stimulus such as light or sound. We can then measure blood flow in different parts of the brain in response to this stimulus, and perform experiments to test whether this response is affected by certain drugs or alterations in conditions such as increased levels of carbon dioxide or alteration in blood flow.If we find that neurovascular coupling exists and is controlled in the same ways in zebrafish as in mammals, our new model would be able to replace the use of protected animals and to provide new scientific insights into neurovascular coupling.

大脑的血液供应由称为“神经血管耦合”的过程紧密控制。这增加了大脑区域的血液供应，尽管如何实现这一目标。神经血管耦合在中风和痴呆等疾病中有缺陷，并且了解这一过程可能最终导致改善这些疾病的策略。由于目前只能在活着的大脑中研究具有完整的血液供应的神经血管耦合，因此其研究需要动物的使用，通常需要小鼠和大鼠。这些实验需要手术才能暴露大脑，并在动物中对大脑进行成像，这些动物要么清醒，约束或麻醉，这不可避免地会引起困扰和痛苦。因此，我们希望开发一个模型，可以为神经血管耦合提供科学见解，但减少了侵入性动物实验的数量。斑马鱼在研究神经血管耦合方面具有两个关键优势。首先，因为它是透明的，并且我们可以生成“转基因”动物，其中某些细胞会荧光不同，因此我们可以在没有任何仪器的情况下对大脑进行成像。其次，我们可以在不死的情况下停止心脏，因为它从周围的水中获得了足够的氧气。斑马鱼胚胎在年龄大于5d之前才被认为是一种自由活跃的动物，并且使用这些动物不被认为是政府立法的动物实验（称为非保护）。我们将利用这些优势来开发一种神经血管耦合模型，以在非保护Zebraf胚胎中。我们已经产生了转基因的斑马鱼，当这些细胞中的钙水平增加时，血管或神经荧光明亮。由于钙的水平增加了“激活”的细胞，因此这些转基因线使我们能够成像响应刺激（例如光或声音）激活大脑的哪些部分。然后，我们可以根据这种刺激来测量大脑不同部位的血液流动，并执行实验以测试该反应是否受某些药物的影响或诸如诸如增加二氧化碳水平或血液流动水平的变化之类的变化的影响，如果我们发现神经血管耦合的存在，并且在Zeemals中的使用方式与乳腺癌中的动力相同，可以用来替换乳腺癌，并且能够以新的方式控制疗法，从而可以替换疗法的疗法，并且可以替换疗法的疗程进入神经血管耦合。