Comparison of three cellular processing approaches for studying voltage-gated sodium ion channels

Objective To establish and compare three cellular processing approaches for voltagegated sodium ion channels in which patch clamp technique is suitable to apply.Methods Cells were processed with acute isolation technique,brain slices separating technique and pCMV-SCN1A plasmid calcium-transfected HEK293 cells.Morphology was observed under microscopic vision and activation of sodium ion channel was monitored using patch clamp technique.Results The acute isolated neurons had normal morphology and relatively long processes.The success rate of patch clamp technique was over 50％,with the recorded cellular membrane capacity of (7.56± 3.47)pF (n=20) and serial resistence of (10.18±4.82) MΩ (n=20).Isolated neurons in brain slices were normal and plump.The success rate of patch clamp technique was over 30％,with the recorded cellular membrane capacity of (9.45±4.26)pF (n=10) and serial resistence of (12.53 ± 3.87)MΩ (n=10).Calcium-transfected cells were integrated and stereo.The success rate of patch clamp technique was approximately 20％,with the recorded cellular membrane capacity of (8.79±4.54) pF (n=10) and serial resistence of (14.12±4.26) MΩ (n=10).All these approaches could provide the current of sodium channel required for research.Conclusion These three cellular processing approaches,which are feasible for distinct researches due to different strengths and weaknesses,can record voltage-gated sodium ion channels conveniently and rapidly. Key words: Sodium channels ;  Patch-clamp technique;  Whole-cell recordings;  Epilepsy