Unit

UNIT 2.5 Analysis of Early Gene Responses

  1. Ronald S. Duman

Published Online: 1 MAY 2001

DOI: 10.1002/0471141755.ph0205s00

Current Protocols in Pharmacology

Current Protocols in Pharmacology

How to Cite

Duman, R. S. 2001. Analysis of Early Gene Responses. Current Protocols in Pharmacology. 00:2.5:2.5.1–2.5.20.

Author Information

  1. Yale University School of Medicine and Connecticut Mental Health Center, New Haven, Connecticut

Publication History

  1. Published Online: 1 MAY 2001
  2. Published Print: MAR 1998

Abstract

Early response genes, also referred to as immediate early genes, are rapidly induced in response to a variety of extracellular stimuli, and have been used as markers of cellular activation. Expression of immediate early genes, such as c-fos, can be used to determine the influence of specific drugs on receptor regulation of intracellular signaling pathways. This unit describes two techniques for the analysis of mRNA from immediate early genes. Northern blot analysis provides a method for size-fractionation of native mRNA by gel electrophoresis and visualization of a specific mRNA in a pool of total RNA isolated from a tissue of interest. Thus, this analysis provides important information on the size and abundance of the mRNA of interest and on the specificity of the hybridization probe. However, this approach does not provide information on the cellular localization of the mRNA. Such data may be obtained by in situ hybridization of mRNA in tissue sections. In addition to the northern and in situ hybridization techniques, this unit provides two support protocols for radiolabeling either RNA or DNA probes along with information on the selection of an appropriate probe and hybridization conditions.