Establishment of a method to isolate sinoatrial node cells of adult rats and observation of the expression of sodium subtypes
10.7652/jdyxb201706025
- VernacularTitle:成年大鼠窦房结活细胞分离方法的建立及钠通道各亚型表达的观察
- Author:
juan Ya NI
1
;
yuan Hong BAI
;
wen Jing ZHANG
;
dong Guo YANG
;
qun Ai MA
Author Information
1. 西安交通大学第二附属医院心血管内科
- Keywords:
rat;
sinoatrial node cell;
isolation;
sodium channel;
expression
- From:
Journal of Xi'an Jiaotong University(Medical Sciences)
2017;38(6):912-916,925
- CountryChina
- Language:Chinese
-
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.
