MOLECULAR PROFILE OF LAMINA-SPECIFIC ALTERATIONS IN THE DLPFC IN SCHIZOPHRENIA

精神分裂症 DLPFC 层特异性改变的分子谱

• 批准号: 7553453
• 负责人: Karoly Mirnics
• 金额: $ 18.65万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-07-01 至 2008-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7553453
• 关键词: Affect; Alkaline Phosphatase; Area; Biological Markers; Collaborations; DNA Microarray Chip; DNA Microarray format; Data; Depth; Dissection; Exhibits; Figs - dietary; Gene Expression; Gene Expression Alteration; Genes; Harvest; Human; Impaired cognition; In Situ Hybridization; Individual; Interneurons; Investigation; Label; Lasers; Latex Bead; Link; Localized; Location; Microscopy; Molecular; Molecular Profiling; Monkeys; Neuroglia; Neurons; Numbers; Parvalbumins; Pattern; Population; Population Projection; Prefrontal Cortex; Pyramidal Cells; Schizophrenia; Technology; Testing; Transcript; base; cDNA Arrays; cell type; density; design; gamma-Aminobutyric Acid; hippocampal pyramidal neuron; interest; mRNA Expression; research study; selective expression

As summarized in the Center Overview, abnormalities in the circuitry of the dorsolateral prefrontal cortex (DLPFC) in schizophrenia may be more prominent in layer 3 than in other cortical layers. For example, DLPFC layer 3 pyramidal neurons exhibit somatodendritic morphological abnormalities, and these changes may preferentially affect a subpopulation of pyramidal neurons in this laminar location (see Project 1-Lewis). In order to explore the molecular counterpart to the morphological alterations observed in layer 3, we propose to test the following three hypotheses: 1) DNA microarrays can uncover multiple expression alterations in DLPFC layer 3 of subjects with schizophrenia that are not evident when all layers are analyzed together; 2) Some expression differences observed in layer 3 of subjects with schizophrenia are specific for subpopulations of projection neurons; and 3) Molecular markers that show expression alterations in subjects with schizophrenia are present in pyramidal neurons that share a common projection target. These hypotheses, which are complementary to those proposed in Project 1-Lewis, will be assessed in the following three specific aims. In Aim 1, using high density cDNA microarrays, we will compare deep layer 3 transcriptomes to whole DLPFC harvests in control subjects. This will identify genes that show expression enrichment in DLPFC deep layer 3. Following this, using the same microarrays and laser dissection microscopy we will identify mRNA expression differences in DLPFC layer 3 between pairs of subjects with schizophrenia and matched controls. In Aim 2 we will determine the cellular localization of the most promising expression changes using in situ hybridization and identify markers that show altered transcript levels in a subpopulation of projections neurons. In Aim 3 we will determine whether the subpopulations of DLPFC pyramidal neurons affected in schizophrenia share common projection targets by combining retrograde tracing of monkey DLPFC neurons with in situ hybridization using markers validated in Aim 2. These investigations have a number of conceptual and technical links with other Center projects and depend upon support provided by all of the proposed cores.

正如中心概述中总结的那样，与其他皮质层相比，精神分裂症中背外侧前额叶皮层（DLPFC）的异常可能更为突出。例如，DLPFC第3层锥体神经元表现出体状形态学异常，这些变化可能会优先影响该层流位置中金字塔神经元的亚群（请参阅项目1-LEWIS）。为了探索分子 与在第3层观察到的形态变化的对应物相对于以下三个假设来测试以下三个假设：1）DNA微阵列可以发现患有精神分裂症的受试者的DLPFC第3层的多个表达变化，而当所有层一起分析时，这些层次都不明显； 2）在精神分裂症患者的第3层中观察到的一些表达差异是针对投影神经元亚群的； 3）显示的分子标记 具有共同投射靶标的锥体神经元中存在精神分裂症受试者的表达改变。这些假设将在以下三个特定目标中评估，这些假设将与项目1-Lewis中提出的假设进行互补。在AIM 1中，使用高密度cDNA微阵列，我们将比较深层3个转录组与对照受试者中的整个DLPFC收获。这将识别显示表达富集的基因 在DLPFC深3层中。此后，使用相同的微阵列和激光解剖显微镜，我们将在具有精神分裂症和匹配对照对象的受试者对之间确定DLPFC层3中DLPFC层中的mRNA表达差异。在AIM 2中，我们将使用原位杂交确定最有希望的表达变化的细胞定位，并确定在投影神经元亚群中显示出变化的转录水平的标记。目标 3我们将确定在精神分裂症中受影响的DLPFC锥体神经元的亚群是否通过将猴子DLPFC神经元的逆行追踪与AIM 2验证的原位杂交相结合，共享共同的投影目标，这些研究在AIM 2中验证了。这些研究与其他概念和技术链接在其他中心项目上有许多概念和技术链接。