An improved method for acute isolation of neurons from the hippocampus of adult rats suitable for patch-clamping study.
An improved method is described for fast and reliable isolation of neurons from hippocampus of adult rats by a combination of mechanical and enzymatic means. The procedure allows the isolation of neurons from 500-600-d-old rats (over 300 g), preserving the proximal dendritic structure without impairing the electrical characteristics of the cells. Morphologically distinct neurons can be recognized. Using cell-attached, inside-out and whole-cell configurations of patch clamp technique, it was shown that the enzymatically isolated neurons in hippocampus from rats weighing more than 300 g exhibited voltage-gated calcium, sodium and potassium currents, outwardly rectifying chloride channel and large conductance Ca(2+)-activated potassium channel currents. Approximately, 95% of healthy cells allowed the formation of giga-ohm seals.
Li
Xiao-Ming
XM
Department of Anatomy, The First Military Medical University, Guangzhou 510515 China.
Li
Jian-Guo
JG
Yang
Jian-Ming
JM
Hu
Ping
P
Li
Xiao-Wen
XW
Wang
Ying
Y
Qin
Lu-Ning
LN
Gao
Tian-Ming
TM
D000818:Animals; D002469:Cell Separation / Q000379:methods*; D006624:Hippocampus / Q000166:cytology*; D015640:Ion Channel Gating / Q000502:physiology; D009474:Neurons / Q000166:cytology; D018408:Patch-Clamp Techniques; D024681:Potassium Channels, Calcium-Activated / Q000502:physiology; D051381:Rats; D017208:Rats, Wistar
D024681:Potassium Channels, Calcium-Activated