Detection of gene expression in single neurons by patch-clamp and single-cell reverse transcriptase polymerase chain reaction

被引:17
作者
Chiang, LW [1 ]
机构
[1] Stanford Univ, Med Ctr, Dept Neurobiol, Stanford, CA 94305 USA
关键词
neuron; nitric oxide synthase; heme oxygenase; synaptic plasticity; calcium signaling;
D O I
10.1016/S0021-9673(98)00156-3
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Detection and quantitation of gene expression in single cells is especially important in the central nervous system where, at the cellular level, the synapse can be considered the single functional unit. For example, the consolidation of long-term memories may be mediated by persistent changes in the strength of synaptic transmission at individual synapses. In order to investigate the requirement for de novo RNA synthesis during long-term potentiation in individual neurons, we have combined single-cell electrophysiology with single-cell gene-expression methodology. Described are methods combining whole-cell patch-clamp and single-cell RT-PCR for the detection of a single mRNA species for nitric oxide synthase, or, through a multiplex strategy, for the simultaneous detection of several mRNAs including heme oxygenase 2, protein phosphatase inhibitor 1 protein, and several isoforms of the calcium/calmodulin dependent protein kinase II. (C) 1998 Pubished by Elsevier Science B.V.
引用
收藏
页码:209 / 218
页数:10
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