Jansen Hailey J, Rose Robert A
Libin Cardiovascular Institute of Alberta, Department of Cardiac Sciences, Department of Physiology and Pharmacology, Cumming School of Medicine, University of Calgary.
Libin Cardiovascular Institute of Alberta, Department of Cardiac Sciences, Department of Physiology and Pharmacology, Cumming School of Medicine, University of Calgary;
J Vis Exp. 2019 Jul 25(149). doi: 10.3791/59588.
The electrophysiological properties of atrial myocytes importantly affect overall cardiac function. Alterations in the underlying ionic currents responsible for the action potential can cause pro-arrhythmic substrates that underlie arrhythmias, such as atrial fibrillation, which are highly prevalent in many conditions and disease states. Isolating adult mouse atrial cardiomyocytes for use in patch-clamp experiments has greatly advanced our knowledge and understanding of the cellular electrophysiology in the healthy atrial myocardium and in the setting of atrial pathophysiology. In addition, studies using genetic mouse models have elucidated the role of a vast array of proteins in regulating atrial electrophysiology. Here we provide a detailed protocol for the isolation of cardiomyocytes from the atrial appendages of adult mice using a combination of enzymatic digestion and mechanical dissociation of these tissues. This approach consistently and reliably yields isolated atrial cardiomyocytes that can then be used to characterize cellular electrophysiology by measuring action potentials and ionic currents in patch-clamp experiments under a number of experimental conditions.
心房肌细胞的电生理特性对整体心脏功能有重要影响。负责动作电位的潜在离子电流的改变可导致心律失常的促心律失常基质，如心房颤动，心房颤动在许多情况和疾病状态中非常普遍。分离成年小鼠心房心肌细胞用于膜片钳实验，极大地推进了我们对健康心房心肌和心房病理生理学背景下细胞电生理的认识和理解。此外，使用基因小鼠模型的研究阐明了大量蛋白质在调节心房电生理中的作用。在这里，我们提供了一个详细的方案，用于使用酶消化和这些组织的机械解离相结合的方法，从成年小鼠的心房附属物中分离心肌细胞。这种方法始终如一地可靠地产生分离的心房心肌细胞，然后可用于通过在多种实验条件下的膜片钳实验中测量动作电位和离子电流来表征细胞电生理。
