WP03 - Gene Identification

The objective of WP3 is the generation and validation of candidate genes in a hypothesis-free approach.

Brain tissue from rats showing extremes in impulsive and reward-related behaviour (one deliverable from WP1) will be studied for differential gene expression in relevant brain regions such as the nucleus accumbens, prefrontal cortex and amygdala. For screening purposes, an Affymetrix platform and a Serial Analysis of Gene Expression (SAGE) platform will be used. Our research consortium set up a patented micro-method for transcriptome characterization using the SAGE technique. An in-house generated database consisting of > 1.2 million SAGE tags sampled from the rat brain will be screened to identify genes preferentially expressed in brain regions involved in reward-related behavioural traits. Furthermore, a highly standardized Affymetrix platform will be used for the search of candidate genes. Selected genes will be confirmed with an independent method namely realtime RT-PCR (SYBR green or Taq-Man). Confirmed, selected genes will be further analysed by in situ hybridization and if possible selected genes will be also evaluated on the protein level by Western blotting and immunohistochemistry.

WP leader is Professor Dr. Rainer Spanagel. Rainer Spanagel is the head of the Department Psychopharmacology, CIMH. He is working on drug-induced reward processes since 1989 and has published more than 100 peer-reviewed papers. He is Co-ordinator of the NGFN program Genetics on Alcoholism and has combined his behavioural work with gene expression profiling in a very successful EU funded project (TARGALC; 5th EU-program).



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