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To establish a method to isolate single sinoatrial node cells of adult rats to observe the cells morphology structure and the expression of sodium channel subtype, and to provide experimental bases for the further research on the effects of sodium channel in sinoatrial node function.
To isolate adult rat sinoatrial node tissue and cut it into slices about 2mm width, digest the slices with type IIcollagenase combinate protease, and observe the cells morphology structure, then perform immunofluorescence staining with HCN4 and laser confocal imaging to identify cells and observe the expression of Nav1.1、Nav1.2、Nav1.3、Nav1.5、Nav1.6、Nav1.7、Nav1.8、Nav1.9.
The shape of the isolated cells is curved, spindle, and slender, but the main is spindle-shaped. The stripes of cells are clear, have good bioactivity and can survive 6-8 hours; Sodium channel Nav1.1、Nav1.5、Nav1.6、Nav1.7、Nav1.8、Nav1.9 are positive expression in adult rat sinoatrial node cells membrane without nucleus, but Nav1.2、Nav1.3 are negative expression.
The method of type IIcollagenase combinate protease to digest and isolate adult rat sinoatrial node cells is reliable, and Sodium channel subtypes are differently expressed in sinoatrial node cells.