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Review : Single-Cell mRNA Amplification: Implications for Basic and Clinical Neuroscience

Department of Pharmacology, Department of Psychiatry, University of Pennsylvania Medical School

Peter Crino

Department of Pharmacology, Department of Neurology, University of Pennsylvania Medical School

Marc Dichter

Department of Pharmacology, Department of Neurology, University of Pennsylvania Medical School, The Graduate Hospital Philadelphia, Pennsylvania

The application of molecular biological techniques to the study of single cells has provided a unique window for exploring the mechanisms that underlie integrated cellular function. Analysis of gene expression in indi vidual cells of the central nervous system is critical to understanding how distinct cell populations with differing chemical and anatomic phenotypes respond to pharmacological agents or are altered in disease states. Quantification of mRNA by single-cell analysis gives a high-resolution picture of changes in gene expression within individual cells, whereas more conventional types of mRNA analysis may obscure subtle changes in gene expression because of a lack of change in surrounding cells that are included in the mRNA sample. In addition, the sensitivity for detecting low levels of mRNA is enhanced when individual versus groups of cells are analyzed. With the advent of various mRNA amplification strategies, it is now possible to determine the mRNA composition or "expression profile" of individual cells. Information about relative levels of different mRNAs, the subcellular localization of mRNAs, and insight into cell-specific RNA splicing and RNA editing can be obtained. When these molecular data are combined with electrophysiological, morpho logical, immunohistochemical, and anatomical analyses, a detailed portrait of neuronal functioning can be obtained. Moreover, alterations in cellular functioning induced by physiological manipulation, drug adminis tration, or disease state can be monitored by combining these approaches. This precise cellular information may be useful in developing pharmaceuticals designed to alter mRNA levels or protein levels in a predictable manner (transcript-aided drug design) to elicit specific physiological states. The Neuroscientist 1:200-211, 1995

Key Words: KEY WORDS aRNA amplification • PCR • Expression profile • Transcript-aided drug design

The Neuroscientist, Vol. 1, No. 4, 200-211 (1995)
DOI: 10.1177/107385849500100404


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