Patch-clamp recording of amino acid-activated responses in "organotypic" slice cultures.
Patch-clamp recording of amino acid-activated responses in "organotypic" slice cultures.
复制标题
膜片钳记录“器官型”切片培养物中氨基酸激活的反应。
DOI：
10.1073/pnas.85.9.3221
复制
发表时间：
1988
影响因子：
11.1
通讯作者：
Gähwiler,BH
中科院分区：
文献类型：
--
作者：
Llano,I;Marty,A;Johnson,JW;Ascher,P;Gähwiler,BH
膜片钳记录技术用于研究大鼠小脑和海马培养的“器官型”切片中氨基酸激活通道的特性。海马锥体细胞对三种主要的谷氨酸能激动剂：N-甲基-D-天冬氨酸（N-Me-D-Asp）、使君子酸盐和红藻氨酸有反应，而浦肯野细胞仅对使君子酸盐和红藻氨酸有反应。对海马神经元外向斑块中记录的单通道事件的分析显示出大的电导事件（50 pS），这种事件在甘氨酸存在的情况下更频繁地发生。这些事件可能由 N-Me-D-Asp 产生，也可能由使君子酸产生（频率较低）。另一方面，在浦肯野神经元中从未观察到 50-pS 事件。这支持了 N-Me-D-Asp 和“非 N-Me-D-Asp”受体是不同分子实体的假设。浦肯野细胞的全细胞和外侧斑片记录的比较揭示了γ-氨基丁酸（GABA）和谷氨酸受体的明显空间分离：虽然GABA受体在体膜中以高密度存在，但使君子酸和红藻氨酸受体大多位于体外。结果表明，器官切片培养物适用于膜片钳方法。他们还表明，在这些培养物中，氨基酸受体根据细胞类型和细胞内区域具有特定的分布模式。
Patch-clamp recording techniques were used to study the properties of amino acid-activated channels in cultured "organotypic" slices from rat cerebellum and hippocampus. Hippocampal pyramidal cells responded to the three main glutamatergic agonists, N-methyl-D-aspartate (N-Me-D-Asp), quisqualate, and kainate, whereas Purkinje cells responded only to quisqualate and kainate. Analysis of single-channel events recorded in outside-out patches from hippocampal neurons showed large conductance events (50 pS), which occurred more frequently in the presence of glycine. These events could be produced by N-Me-D-Asp and also, at low frequency, by quisqualate. On the other hand, 50-pS events were never observed in Purkinje neurons. This supports the hypothesis that N-Me-D-Asp and "non-N-Me-D-Asp" receptors are distinct molecular entities. Comparison of whole-cell and outside-out patch recordings from Purkinje cells revealed a clear spatial segregation of gamma-aminobutyric acid (GABA) and glutamate receptors: although GABA receptors are found at high density in somatic membrane, quisqualate and kainate receptors are mostly extrasomatic. The results show that organotypic slice cultures are amenable to patch-clamp methods. They also show that, in these cultures, amino acids receptors have specific distribution patterns according to cell type and to region within a cell.
影响因子：
5.4
作者：
G. Shepherd
通讯作者：
G. Shepherd