Dubach J Matthew, Das Saumya, Rosenzweig Anthony, Clark Heather A
Biomedical Engineering Group, The Charles Stark Draper Laboratory, 555 Technology Square, Cambridge, MA 02139, USA.
Proc Natl Acad Sci U S A. 2009 Sep 22;106(38):16145-50. doi: 10.1073/pnas.0905909106. Epub 2009 Sep 3.
Regulation of sodium flux across the cell membrane plays a vital role in the generation of action potentials and regulation of membrane excitability in cells such as cardiomyocytes and neurons. Alteration of sodium channel function has been implicated in diseases such as epilepsy, long QT syndrome, and heart failure. However, single cell imaging of sodium dynamics has been limited due to the narrow selection of fluorescent sodium indicators available to researchers. Here we report, the detection of spatially defined sodium activity during action potentials. Fluorescent nanosensors that measure sodium in real-time, are reversible and are completely selective over other cations such as potassium that were used to image sodium. The use of the nanosensors in vitro was validated by determining drug-induced activation in heterologous cells transfected with the voltage-gated sodium channel Na(V)1.7. Spatial information of sodium concentrations during action potentials will provide insight at the cellular level on the role of sodium and how slight changes in sodium channel function can affect the entirety of an action potential.
跨细胞膜的钠通量调节在动作电位的产生以及心肌细胞和神经元等细胞的膜兴奋性调节中起着至关重要的作用。钠通道功能的改变与癫痫、长QT综合征和心力衰竭等疾病有关。然而,由于研究人员可用的荧光钠指示剂选择范围狭窄,钠动力学的单细胞成像一直受到限制。在此我们报告,在动作电位期间对空间定义的钠活性进行检测。实时测量钠的荧光纳米传感器是可逆的,并且对用于成像钠的其他阳离子(如钾)具有完全选择性。通过确定在转染了电压门控钠通道Na(V)1.7的异源细胞中药物诱导的激活,验证了纳米传感器在体外的使用。动作电位期间钠浓度的空间信息将在细胞水平上提供关于钠的作用以及钠通道功能的微小变化如何影响整个动作电位的见解。