Aziz Qadeer, Nobles Muriel, Tinker Andrew
Clinical Pharmacology, William Harvey Research Institute, Barts and the London School of Medicine and Dentistry, Queen Mary University of London, London, UK.
Bio Protoc. 2020 Jan 5;10(1):e3477. doi: 10.21769/BioProtoc.3477.
The cardiac conduction system allows the synchronized propagation of electrical activity through heart muscle. This is initiated by the spontaneous activity of the specialized pacemaker cells of the sino-atrial node (SAN). The SAN region underlies automaticity in mammals and therefore has a crucial role in the pathogenesis of cardiac disorders such as arrhythmia. Isolation of SAN tissue and SAN cells is critical to advance our understanding of SAN structure and function in health and disease. Initially, isolation of SAN tissue and SAN cells was carried out in the rabbit owing to its larger size and similar electrical properties to human. This protocol was optimized by Mangoni and Nargeot (2001) for use in mice to take advantage of advancements in transgenic models. Here, we provide a step-by-step guide to dissecting the SAN tissue and isolating pacemaker cardiomyocytes from mouse hearts using an enzyme digestion approach.
心脏传导系统使电活动能够在心肌中同步传播。这一过程由窦房结(SAN)中特殊起搏细胞的自发活动启动。在哺乳动物中,窦房结区域是自动节律性的基础,因此在心律失常等心脏疾病的发病机制中起着关键作用。分离窦房结组织和窦房结细胞对于增进我们对其在健康和疾病状态下的结构与功能的理解至关重要。最初,由于兔子体型较大且电特性与人类相似,窦房结组织和窦房结细胞的分离工作是在兔子身上进行的。Mangoni和Nargeot(2001年)对该方案进行了优化,以便用于小鼠,以利用转基因模型方面的进展。在此,我们提供一份使用酶消化法从小鼠心脏中解剖窦房结组织并分离起搏心肌细胞的分步指南。