Pyramidal neuron hypoactivity in schizophrenia cortex

精神分裂症（SZ）影响世界人口的1%。肌营养不良短蛋白结合蛋白的等位基因变体 1（DTNBP1）基因，现在是SZ的首选候选基因，与健康人的纤维束损伤有关 以及青春期前的脑容量减少都是SZ症状然而，下游影响减少， 对dysbindin知之甚少。功能失调的Dysbindin降低铜转运P型ATP酶 （ATP 7A）和铜转运蛋白CTR 1。ATP 7A和CTR 1促进铜在血液之间的转运 和大脑铜在SZ中的作用一直存在争议，许多研究发现， 患者体内的铜然而，这些研究并没有考虑到铜可能会在血液中积聚，因为它 无法进入大脑。铜减少实验操作产生脱髓鞘，多巴胺 增加，和SZ样行为障碍。总之，dysbindin/铜改变可导致SZ- 像病理学如受损的白色物质完整性、过量的多巴胺和改变的线粒体活性， 然而这种结合途径仍然在很大程度上未被研究。总的来说，我假设铜转运改变 有助于SZ病理学，可能通过降低dysbindin表达。我会在尸体上测试这个 SZ脑、首次发作抗精神病药物初治SZ患者和dysbindin敲除小鼠模型。 具体目标1：我已经学习了蛋白质印迹法，现在正在学习免疫组化分析。 具体目标2A.我们将测试死后黑质和海马体的铜状态， 这种状态与处理和未处理SZ中的铜转运蛋白ATP 7A/CTR 1的关系。此外，委员会认为， 我们将评估抗精神病药物与这些变量相关的作用机制。 具体目标2B。为了发现潜在的精神分裂症生物标志物，我们将评估外周铜状态， 及其与铜转运蛋白和DTNBP1、SLC31A1（CTR 1）和ATP 7A甲基化状态的关系 首次发作、抗精神病药物初治SZ患者接受抗精神病药物治疗6周之前和之后。 具体目标2C。在dysbindin敲除、杂合和野生型小鼠中，我们将测试外周， 海马和黑质铜状态，以及与铜转运蛋白ATP 7A/CTR 1，认知缺陷， 探讨奎替利治疗前后抗精神病药物介导的变化及其机制。 具体目标3：这些研究将提供SZ脑铜状态的第一个证据，并解决 SZ血液中过量铜与铜缺乏引起的SZ样症状之间的并列关系。 此外，这些研究将为风险基因dysbindin及其对铜的影响提供新的数据 转运蛋白（以前未研究的途径在SZ）和抗精神病药物抢救的潜在机制， 铜饥饿缺陷，这可能会产生新的药物开发目标。我将学习定量RT- PCR、甲基化测定、免疫组织化学、电子显微镜和组织切片、脑收集， 科学写作，实验室和动物管理，以及所有这些技能需要，以及熟练的教学。

Schizopphrenia (SZ) affects 1% of the world population. Allelic variants of the dystrobrevin binding protein 1 (DTNBP1) gene, now a top SZ candidate gene, are associated with fiber tract impairments in healthy people and reduced brain volume in preteens- both SZ symptoms. However, the downstream effects of decreased dysbindin are poorly understood. Dysfunctional dysbindin decreases the copper-transporting P-type ATPase (ATP7A) and the copper transporter CTR1. ATP7A and CTR1 facilitate copper transport between the blood and the brain. The role of copper in SZ has been controversial, with many studies finding increased blood copper in patients. However, these studies have not considered copper may build up in the blood because it cannot get into the brain. Copper-decreasing experimental manipulations produce demyelination, dopamine increases, and SZ-like behavioral impairments. Taken together, dysbindin/copper alterations can result in SZ- like pathology such as impaired white matter integrity, excess dopamine, and altered mitochondrial activity, and yet this combined pathway remains largely unstudied. Overall, I hypothesize copper transport alterations contribute to SZ pathology, potentially through decreased dysbindin expression. I will test this in postmortem SZ brain, first-episode antipsychotic-naïve SZ patients, and a dysbindin knockout mouse model. Specific Aim 1: I have learned western blotting, and am now learning immunohistochemical analysis. Specific Aim 2A. We will test the copper state of postmortem substantia nigra and hippocampus, and the relationship of this state to copper transporters ATP7A/CTR1 in both treated and untreated SZ. Furthermore, we will assess the mechanism of action of antipsychotics in relation to these variables. Specific Aim 2B. To discover potential schizophrenia biomarkers, we will assess peripheral copper state and its relationship to copper transporters and methylation status of DTNBP1, SLC31A1(CTR1), and ATP7A before and after 6 weeks of antipsychotic treatment in first-episode, antipsychotic-naïve SZ patients. Specific Aim 2C. In dysbindin knockout, heterozygous, and wild-type mice, we will test peripheral, hippocampal, and nigral copper state, and the relation to copper transporters ATP7A/CTR1, cognitive deficits, and the mechanism of antipsychotic mediated changes before and after 28 days quetiapine treatment. Specific Aim 3: These studies will provide the first evidence of SZ brain copper state, and resolve the juxtaposition between excess copper in SZ blood and the SZ-like symptoms induced by copper deficit. Furthermore, these studies will provide new data about risk gene dysbindin and its effects on copper transporters (a previously unstudied pathway in SZ) and a potential mechanism of antipsychotic rescue of copper starvation deficits, which could yield novel targets for drug development. I will learn quantitative RT- PCR, methylation assay, immunohistochemistry, electron microscopy and tissue sectioning, brain collection, scientific writing, lab and animal management, and all that these skills entail, as well as proficient teaching.