Gene profiling of laser-microdissected brain regions and sub-regions.
The application of transcriptomics and proteomics approaches to accurately dissected anatomically-defined brain regions and sub-regions remains a central focus of current neurobiological investigations as well as a necessary step towards single-neuron neurogenomics and neuroproteomics. A protocol is described for the simple, rapid, and reproducible laser microdissection of brain regions and sub-regions for microarray-based gene expression analyses from individual rats or mice using two rounds of in vitro transcription (IVT). The results presented also demonstrate that the current Affymetrix GeneChip arrays are well suited for this experimental design with high reproducibility and limited effects of the shortening of target RNA caused by the double IVT approach.
Sanna
Pietro Paolo
PP
Department of Neuropharmacology, The Scripps Research Institute, La Jolla, CA 92037, USA. psanna@scripps.edu
King
Alvin R
AR
van der Stap
Lena D
LD
Repunte-Canonigo
Vez
V
D000818:Animals; D001921:Brain / Q000502:physiology*; D020869:Gene Expression Profiling / Q000379:methods*; D007834:Lasers; D042282:Microdissection / Q000295:instrumentation / Q000379:methods*; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D051381:Rats