Uncovering complex expression patterns in schizophrenia

描述(由申请人提供)：我的目标是获得学科的先进和实用知识，这将有助于将微阵列获得的基因表达数据置于更广泛的生物学背景中。对于对基因组学感兴趣的科学家来说，理想的专业知识包括多个神经科学学科，包括(但不限于)群体遗传学、生物统计学和所研究组织的体视学评估。这些额外的专业知识将使我能够识别数据集之间的复杂关系，根据现有的连锁研究解释获得的微阵列数据，对我们的尸检样本进行有限的基因分型实验，并使用无偏见的体视学采样方法验证未发现的基因表达变化。 精神分裂症患者的前额叶皮质(PFC)功能障碍与工作记忆障碍有关，而颞叶上回(STG)的功能改变与精神病有关。此外，不同性别的精神分裂症的表现与发病年龄、症状病理学、病前病史、神经成像结果、药物反应和大脑结构的差异有关。 这项应用围绕两个关键问题：1)在不同的大脑区域内和跨不同的大脑区域是否存在与精神分裂症相关的表达谱；以及2)与精神分裂症相关的表达变化是否因性别而异？在此背景下，我们建议使用三个特定目标来检验七个特定假设：AIM1比较12名男性精神分裂症受试者和匹配对照的前额叶(PFC)和颞叶上皮层(STG)的基因表达模式。目的2.比较12例女性精神分裂症患者和对照组前额叶(PFC)和颞叶上皮质(STG)的基因表达谱。AIMS 1和2将使用相同的方法，并比较转录本：A)使用全基因组HG_U133A和B Affymetrix微阵列。B)使用定制的高灵敏度聚合物EDNA微阵列。这些EDNA聚合物阵列，包括我们的专利探针(专利申请正在进行中)，将使我们能够改进和有针对性地评估许多转录文本，这些转录文本太稀疏，无法被目前可用的微阵列检测到。目的3.利用原位杂交在转录水平上验证和定位微阵列未发现的基因表达变化到细胞类型A)；使用免疫组织化学在蛋白质水平上验证和定位；以及C)通过身体样本的基因分型来跟踪最有希望的表达变化。

DESCRIPTION (provided by applicant): 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.