原代培养海马神经元的多种电压门控钙通道电流成分

Diverse components of voltage-gated calcium channel currents in primary cultured hippocampal neurons

本文旨在探讨原代培养的海马神经元上电压门控钙通道电流(voltage-gated calcium channel currents,VGCCs)的记录和分离。本文建立了一种稳定、高效的记录海马神经元VGCCs的海马神经元培养体系。此法获得的成熟的海马神经元克服了急性分离的海马神经元的缺陷,细胞破膜后20min电流不发生明显衰减。应用全细胞膜片钳技术证实,在本实验条件下,不损伤神经细胞膜的电学特性,可成功地记录到海马神经元的多种VGCCs成分(如L,N,P/Q,T等)。

The present study was to record and separate the voltage-gated calcium channel currents （VGCCs） in primary cultured hippocampal neurons. An improved method is described for efficient and stable recording of VGCCs from primary cultured hippocampal neurons. The procedure allows the obtained hippocampal neurons overcome the defects from acutely isolated hippocampal neurons and currents do not rundown apparently after 20 min of membrane rupture. Using whole-cell configurations of patch clamp technique, it is shown that the method does not damage the neuron membrane electrical properties, and successfully records a variety of VGCCs components in hippocampal neurons （such as L, N, P/Q, T, etc.）.