Biomedical Engineering Reference
In-Depth Information
Chapter 9
Detecting Neural Activity-Dependent
Immediate Early Gene Expression
in the Brain
Kazuhiro Wada , Chun-Chun Chen , and Erich D. Jarvis
Abstract
In this chapter, we present an in situ hybridization protocol with
radioactive probe that has been successfully and easily used on detecting mRNA
expression level and patterns, in multiple tissue types and developmental stages. To
detect behaviorally regulated, i.e., motor or sensory, mRNA expression of immedi-
ate early genes (IEGs) within cells and tissues in vivo, in situ hybridization is a
powerful method for discovering neural activity correlations and novel neural struc-
tures. Compared with nonradioactive probe methods such as digoxigenin (DIG)
labeling, the radioactive probe hybridization method provides a semi-linear relation
between signal intensity and targeted mRNA amounts for quantitative analysis.
Furthermore, this method allows us high-throughput mRNA expression analysis for
100-200 sides with 400-1,000 tissue sections simultaneously. This method allows
identifying the possible signifi cance and function of interested genes in the nervous
system under specifi c behaviors.
Keywords
Activity-dependent gene expression High-throughput in situ hybrid-
ization Immediate early genes (IEGs) Radioactive in situ hybridization Songbird
Vocalization
9.1
Introduction
In nature, many types of behaviors are observed in a wealth of animal species at
specifi c conditions and at specifi c developmental stages with species-specifi c fea-
tures. To understand the neural basis of such behaviors, one critical step is to
