一种改进的适用于膜片钳记录的成年大鼠海马神经元急性分离法.pdfVIP

112 Acta Physiologica Sinica, February 25, 2004, 56(1): 112-117 Experimental Technique An improved method for acute isolation of neurons from the hippocampus of adult rats suitable for patch-clamping study 1,2 2 2 2 2 2 LI Xiao-Ming , LI Jian-Guo , YANG Jian-Ming , HU Ping , LI Xiao-Wen , WANG Ying , QIN Lu-Ning2, GAO Tian-Ming1,2,* Departments of 1Anatomy and 2Physiology, The First Military Medical University, Guangzhou 510515, China Abstract: 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 Ca2+-activated potassium channel currents. Approximately, 95% of healthy cells allowed the formation of giga-ohm seals. Key words: neuron; patch clamp; hippocampus; rat 1,2 2 2 2 2 2 2 1,2,* , , , , , , ,