Quantitative RT-PCR Analysis of Gene Expression Alterations in Selective Brain Regions of Patients with Mood Disorders
Jun Li; P.V. Choudary; S.J. Evans; M.P. Vawter; H. Tomita; J. Meador-Woodruff; E.G. Jones; W.E. Bunney; S.J. Watson; H. Akil; R.M. Myers
52th Annual Meeting of the American Society of Human Genetics. 2002.
We compared transcript levels for several hundred genes in post-mortem brain tissues between patients and controls in an effort to identify gene-expression alterations associated with mood disorders. We isolated total RNA from specific brain regions implicated in mood disorders, synthesized first-strand cDNA, and performed SYBR Green-based realtime-PCR experiments to quantitate transcript levels in each sample relative to a panel of reference genes. So far we have analyzed anterior cingulate cortex, dorsolateral prefrontal cortex and cerebellum from nine Bipolar patients, eleven Major Depressive Disorder (MDD) patients and 20 control individuals. The genes included in this study are approximately 150 carefully selected candidate genes covering a variety of functional categories, additional genes previously reported in literature as putatively associated with suicide, Bipolar and/or MDD, as well as genes that appeared to be up- or down-regulated in our cases as compared to controls according to our Affymetrix microarray analysis (reported elsewhere). This presentation will describe results of our quantitative RT-PCR analysis, as well as our methods for achieving high sensitivity and high throughput, while ensuring accuracy. Supported by: The Pritzker Neuropsychiatric Disorders Research Consortium, Pritzker Family Philanthropic Fund and NIH CONTE Center grant #L99MH60398.