An industrial collaboration to develop new microscopy tools to image ATP usage in muscle

工业合作开发新的显微镜工具来成像肌肉中 ATP 的使用情况

• 批准号: 2873383
• 金额: --
• 依托单位: University of Kent
• 依托单位国家: 英国
• 项目类别: Studentship
• 财政年份: 2023
• 资助国家: 英国
• 起止时间: 2023 至 无数据
• 项目状态: 未结题
• 来源: https://gtr.ukri.org/projects?ref=studentship-2873383
• 关键词: industrial; collaboration; develop; new; microscopy

Cardiac muscle contraction occurs when myosin II in the thick filament uses ATP to power the relative movement of the actincontaining thin filament. Given the vast numbers of myosin motors, if all were active, the heart would be rapidly drained of ATP. Therefore, to save energy, myosin can enter the 'super relaxed (SRX)' state when detached from actin, which has an inhibited ATPase. When needed, e.g., during exercise, these SRX myosins can be reactivated. Hypertrophic cardiomyopathy (HCM), the most common genetic heart disease, is thought to be due to hypercontractility, which may be caused by underpopulation of SRX. We will investigate how SRX is regulated in both relaxed and active conditions. Our approach is to study myosin and actin in their native myofibril environment. This highly ordered structure is capable of contracting in the presence of calcium and ATP. In this project, we will construct a device that uses glass microneedles to grab either end of a myofibril, this will enable the contractile force to be measured. Uniquely, we will combine this with single molecule imaging of ATP being turned over in the myofibrils ofporcine/human cardiac muscle, and stem-cell derived cardiomyocytes. This has not been achieved before and opens up a huge possibility for understanding how muscle is affected by calcium, phosphorylation, force, pharmacological interventions, and disease-associated mutations. By the end of this project, we will provide a comprehensive understanding of the role and regulation of myosin's SRX states in normal and diseased tissue.

当粗肌丝中的肌球蛋白II利用ATP为含肌动蛋白的细丝的相对运动提供动力时，就会发生心肌收缩。鉴于肌球蛋白马达的数量巨大，如果所有马达都处于活跃状态，心脏中的ATP就会迅速耗尽。因此，为了节省能量，肌球蛋白可以进入“超松弛（SRX）”状态时，从肌动蛋白，其中有一个抑制ATP酶。当需要时，例如，在运动过程中，这些SRX肌球蛋白可以被重新激活。肥厚型心肌病（HCM）是最常见的遗传性心脏病，被认为是由于过度收缩，这可能是由SRX的人口不足引起的。我们将研究SRX在放松和活跃条件下是如何调节的。我们的方法是研究肌球蛋白和肌动蛋白在他们的天然肌原纤维环境。这种高度有序的结构能够在钙和ATP存在的情况下收缩。在这个项目中，我们将构建一个设备，使用玻璃微针抓住肌原纤维的两端，这将使收缩力被测量。独特的是，我们将联合收割机与猪/人心肌肌原纤维和干细胞衍生的心肌细胞中ATP翻转的单分子成像相结合。这是以前从未实现过的，为了解肌肉如何受到钙，磷酸化，力，药物干预和疾病相关突变的影响开辟了巨大的可能性。在这个项目结束时，我们将提供一个全面的了解的作用和调节肌球蛋白的SRX状态在正常和病变组织。