ABSTRACT
Objective To establish a method to isolate single sinoatrial node cells of adult rats and observe the cells'morphological structure and the expression of sodium channel subtype so as to provide experimental basis for further research on the role of sodium channel in sinoatrial node function.Methods We isolated adult rat sinoatrial node tissue and cut it into slices about 2 mm in width,digested the slices with type Ⅱ collagenase combinate protease, and observed the cells' morphological structure. We then performed immunofluorescence staining with HCN4 and laser confocal imaging to identify cells and observe the expressions of Nav1 .1 ,Nav1 .2 , Nav1.3,Nav1.5,Nav1.6,Nav1.7,Nav1.8 and Nav1.9.Results The isolated cells had spindle-like,arc-like, and slender and curved shapes,but were mainly spindle-shaped.The stripes of the cells were clear,had good bioactivity and could survive 6-8 hours.Sodium channels Nav1.1,Nav1.5,Nav1.6,Nav1.7,Nav1.8,and Nav1.9 were positively expressed in adult rat sinoatrial node cells,while Nav1 .2 and Nav1 .3 were negatively expressed. Conclusion The method of digesting and isolating adult rat sinoatrial node cells with type Ⅱ collagenase combinate protease is reliable,and sodium channel subtypes are differently expressed in sinoatrial node cells.
ABSTRACT
Objective To establish a method to isolate single sinoatrial node cells of adult rats and observe the cells'morphological structure and the expression of sodium channel subtype so as to provide experimental basis for further research on the role of sodium channel in sinoatrial node function.Methods We isolated adult rat sinoatrial node tissue and cut it into slices about 2 mm in width,digested the slices with type Ⅱ collagenase combinate protease, and observed the cells' morphological structure. We then performed immunofluorescence staining with HCN4 and laser confocal imaging to identify cells and observe the expressions of Nav1 .1 ,Nav1 .2 , Nav1.3,Nav1.5,Nav1.6,Nav1.7,Nav1.8 and Nav1.9.Results The isolated cells had spindle-like,arc-like, and slender and curved shapes,but were mainly spindle-shaped.The stripes of the cells were clear,had good bioactivity and could survive 6-8 hours.Sodium channels Nav1.1,Nav1.5,Nav1.6,Nav1.7,Nav1.8,and Nav1.9 were positively expressed in adult rat sinoatrial node cells,while Nav1 .2 and Nav1 .3 were negatively expressed. Conclusion The method of digesting and isolating adult rat sinoatrial node cells with type Ⅱ collagenase combinate protease is reliable,and sodium channel subtypes are differently expressed in sinoatrial node cells.