与红外可视膜片钳技术相结合的南方鲇面叶脑片急性分离培养技术

探讨南方鲇初级味觉中枢—面叶脑片的急性分离与不同孵育液中体外培养方法,并建立硬骨鱼类脑片可视膜片钳技术,在脑片局部神经回路水平上深入研究鱼类味觉初级中枢神经元在信息编码中的生理功能,阐述鱼类味觉信号识别的机制。结果表明:①温度25℃、2 h以内、HEPES+Vc+TU+kynurenic acid+低钙脑脊液是最佳体外培养条件。②运用可视法脑片膜片钳实验技术发现南方鲇面叶神经元可分为两种类型。生理条件下,多数细胞处于静息状态,无或少频率适应现象。椭圆形比梨形神经元膜阻抗更高,诱发放电频率却较低。

Acute Isolation and Culture of Facial Lobe Slice from Southern Catfish (Silurus meridionalis Chen) Recorded by Infrared Visual Patch Clamp Techenique

The dissociation and culture in different artificial cerebrum spinal fluids (ACSF) of facial lobe slice isolated from Southern catfish (Silurus meridionalis Chen) are described. To observe the physiological functions in information coding of primary gustatory central neurons and to elaborate the mechanism of information identification in fish, we established the teleost infrared visual patch clamp method. According to experimental requirement, low calcium ACSF contained HEPES, Vc, TU and kynurenic acid could be selected for normal brain slices culture at the temperature of 25 ℃within 2 h. The morphology and electrophysiological characteristics of neurons were observed non invasively by brain slice patch clamp with infrared videomicroscopy. There were two types of neurons (pyriform and oval neurons) in facial lobe. They all had no or low rate of spontaneous discharges and frequency adaptation. Pyriform neuron had lower membrane resistance but with higher evoked discharge frequency.