My objective is to acquire advanced and working knowledge of disciplines that will help place the microarray-obtained gene expression data into a broader biological context. For a scientist interested in genomics, ideal expertise encompasses multiple neuroscience disciplines, including (but not limited to) population genetics, biostatistics and stereological assessment of the studied tissue. This additional expertise will enable me to identify complex relationships between datasets, interpret the obtained microarray data in the light of existing linkage studies, perform limited genotyping experiments on our postmortem samples and verify the uncovered gene expression changes using non-biased, stereological sampling methods. Dysfunction of the prefrontal cortex (PFC) in schizophrenia has been associated with deficits of working memory, while functional changes in the superior temporal gyrus (STG) have been related to psychosis. Furthermore, the presentation of schizophrenia across genders has been associated with differences in age of onset, symptomathology, premorbid history, neuroimaging findings, drug responsiveness and brain structure. This application is focused around two critical questions: 1) Is there a schizophrenia-related expression profile within and across different brain regions and 2) Are schizophrenia-related expression changes different across genders? In this context, we propose to test seven specific hypotheses using 3 specific aims:
Aim1 Compare gene expression pattern in 12 MALE subjects with schizophrenia and matched controls across the prefrontal (PFC) and superior temporal (STG) cortices.
Aim 2. Compare gene expression pattern in 12 FEMALE subjects with schizophrenia and matched controls across the prefrontal (PFC) and superior temporal (STG) cortices.
Aims 1 and 2 will share the same methodology, and compare the transcriptomes: A) Using whole genome HG_U133A and B Affymetrix microarrays. B) Using custom-made, high-sensitivity polymer eDNA microarrays. These eDNA polymer arrays, involving our proprietary probes (patent application in progress) will allow us an improved and targeted assessment of many transcripts that are too sparse to be detected by the currently available microarrays.
Aim 3. Verify and localize the microarray-uncovered gene expression changes to cell types A) at transcript level using in situ hybridization; B) at protein level using immunohistochemistry and C) follow up the most promising expression changes with genotyping of postmortem samples.
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