TY - JOUR
T1 - Analysis of gene expression in single live neurons
AU - Eberwine, James
AU - Yeh, Hermes
AU - Miyashiro, Kevin
AU - Cao, Yanxiang
AU - Nair, Suresh
AU - Finnell, Richard
AU - Zettel, Martha
AU - Coleman, Paul
PY - 1992
Y1 - 1992
N2 - The present here a method for broadly characterizing single cells at the molecular level beyond the more common morphological and transmitter/receptor classifications. The RNA from defined single cells is amplified by microinjecting primer, nucleotides, and enzyme into acutely dissociated cells from a defined region of rat brain. Further processing yields amplified antisense RNA. A second round of amplification results in >106-fold amplification of the original starting material, which is adequate for analysis - e.g., use as a probe, making of cDNA libraries, etc. We demonstrate this method by constructing expression profiles of single live cells from rat hippocampus. This profiling suggests that cells that appear to be morphologically similar may show marked differences in patterns of expression. In addition, we characterize several mRNAs from a single cell, some of which were previously undescribed, perhaps due to "rarity" when averaged over many cell types. Electrophysiological analysis coupled with molecular biology within the same cell will facilitate a better understanding of how changes at the molecular level are manifested in functional properties. This approach should be applicable to a wide variety of studies, including development, mutant models, aging, and neurodegenerative disease.
AB - The present here a method for broadly characterizing single cells at the molecular level beyond the more common morphological and transmitter/receptor classifications. The RNA from defined single cells is amplified by microinjecting primer, nucleotides, and enzyme into acutely dissociated cells from a defined region of rat brain. Further processing yields amplified antisense RNA. A second round of amplification results in >106-fold amplification of the original starting material, which is adequate for analysis - e.g., use as a probe, making of cDNA libraries, etc. We demonstrate this method by constructing expression profiles of single live cells from rat hippocampus. This profiling suggests that cells that appear to be morphologically similar may show marked differences in patterns of expression. In addition, we characterize several mRNAs from a single cell, some of which were previously undescribed, perhaps due to "rarity" when averaged over many cell types. Electrophysiological analysis coupled with molecular biology within the same cell will facilitate a better understanding of how changes at the molecular level are manifested in functional properties. This approach should be applicable to a wide variety of studies, including development, mutant models, aging, and neurodegenerative disease.
KW - Amplified, antisense rna
KW - Expression profile
KW - Mrna complexity
KW - Pyramidal cell
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U2 - 10.1073/pnas.89.7.3010
DO - 10.1073/pnas.89.7.3010
M3 - Article
C2 - 1557406
AN - SCOPUS:0026588147
SN - 0027-8424
VL - 89
SP - 3010
EP - 3014
JO - Proceedings of the National Academy of Sciences of the United States of America
JF - Proceedings of the National Academy of Sciences of the United States of America
IS - 7
ER -