Neurocircuitry Mapping and Genotyping Core

神经回路图谱和基因分型核心

• 批准号: 8328777
• 负责人: ROBERT J. HITZEMANN
• 金额: $ 29.49万
• 依托单位: OREGON HEALTH & SCIENCE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2001
• 资助国家: 美国
• 起止时间: 2001-09-27 至 2016-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8328777
• 关键词: Affect; Alcohol consumption; Alcoholism; Alternative Splicing; Amygdaloid structure; Animal Model; Animals; Area; Brain; Brain region; Breeding; Cell Nucleus; Cells; Chronic; Code; Complement; Complex; Confusion; Control Animal; Control Groups; Data; Data Analyses; Detection; Development; Dissection; Ethanol; Funding; Gene Expression; Gene Expression Profile; Genes; Genotype; Grant; Hand; Human; Inbred Strain; Inbreeding; Individual; Laboratories; Lasers; Lead; Libraries; Macaca; Macaca mulatta; Maps; Masks; Microarray Analysis; Modeling; Mus; National Institute on Alcohol Abuse and Alcoholism; Neurosciences; Occipital lobe; Oligonucleotide Microarrays; Pathway Analysis; Predisposition; Procedures; Proteins; RNA Splicing; Reading; Research; Research Personnel; Sample Size; Sampling; Services; Source; Stress; Tissues; Transcript; Weight; Work; alcohol exposure; alcoholism therapy; animal breeding; base; binge drinking; cDNA Arrays; drinking; insight; member; new therapeutic target; next generation; protein function; segregation; tool

DESCRIPTION (provided by applicant): This is a competing renewal application for a U01 grant entitled "Neurocircuitry Mapping and Genotyping Core"; the application is submitted as a member of the NIAAA sponsored "Integrative Neuroscience Initiative on Alcoholism (INIA)-West (G. Koob, PI). The application continues the focus of the current funding period on both research and core activities. Key core activities of the current funding period were a) the mastery of the use of the Weighted Gene Co-variance Network Analysis (WGCNA) for moderate to large sample sizes (lancu et al. 2010) and b) the development of a strategy for and implementation of quantitative RNAseq (Bottomly et al, 2011; Appendix A). With these tools in hand, we propose 1) to directly sequence the transcriptome ( ~ 25,000,000 75 bp reads/sample) in both replicate High Drinking in the Dark (HDID) mouse lines and in the HS/NPT control animals and 2) to sequence the transcriptome HDID animals that have completed the chronic intermittent ethanol (CIE) procedure with the appropriate control groups. The tissues needed for this analysis will be provided by the Crabbe U01. As the HDID and controls are derived from a 8- way inbred strain cross (Hitzemann et al. 1994), RNAseq is particularity useful, given that masking oligonucleotide array data is never optimal (see Walter et al. 2007,2009). N = 32/group; previous work (lancu et al. 2010) has illustrated that samples of this size are adequate for the proposed analyses. Samples are collected by laser capture micro-dissection (LCM); the regional priority for analysis will be the central nucleus of the amygdala (CeA) > the infralimbic cortex (IL) > the prelimbic cortex (PL). The occipital cortex (OC) will be used as a control region. Aim 1 focuses on binge drinking whereas aim 2 focuses on how chronic ethanol exposure affects ethanol consumption in limited access 2-bottle choice paradigm. Our working hypothesis is that differences between co-expression networks and not the differential expression of individual genes have the greatest translational value (see e.g. Oti et al. 2008; Zhao et al. 2010). In Aim 3, samples from ethanol exposed macaques (Grant U01- INlA-Stress) will be sequenced. Data from the CeA and cortical areas 25 and 32 will be compared to the results obtained in specific aims 1 and 2.

描述（由申请人提供）：这是一项 U01 拨款的竞争续展申请，题为“神经电路图谱和基因分型核心”；该申请是作为 NIAAA 赞助的“酗酒综合神经科学计划 (INIA)-West（G. Koob，PI）的成员提交的。该申请继续将当前资助期的重点放在研究和核心活动上。当前资助期的关键核心活动是 a) 掌握对中等到大样本量的加权基因协方差网络分析 (WGCNA) 的使用 (lancu et 等人。 2010）和b）定量 RNAseq 策略的制定和实施（Bottomly 等人，2011；附录 A）。有了这些工具，我们建议 1) 在重复的黑暗中饮酒 (HDID) 小鼠系和 HS/NPT 对照动物中直接对转录组（约 25,000,000 个 75 bp 读数/样本）进行测序，2) 对已完成实验的 HDID 动物的转录组进行测序 慢性间歇性乙醇（CIE）程序与适当的对照组。此分析所需的组织将由 Crabbe U01 提供。由于 HDID 和对照源自 8 路近交系杂交（Hitzemann 等人，1994 年），因此 RNAseq 特别有用，因为掩蔽寡核苷酸阵列数据从来都不是最佳的（参见 Walter 等人，2007，2009）。 N= 32人/组；先前的工作（lancu 等人，2010）表明，这种规模的样本足以进行拟议的分析。通过激光捕获显微切割（LCM）收集样品；分析的区域优先顺序是杏仁核中央核 (CeA) > 边缘下皮层 (IL) > 边缘皮层 (PL)。枕骨 皮质（OC）将用作控制区域。目标 1 侧重于酗酒，而目标 2 侧重于在有限的 2 瓶选择范式中长期接触乙醇如何影响乙醇消耗。我们的工作假设是共表达网络之间的差异而不是单个基因的差异表达具有最大的翻译价值（参见例如 Oti 等人 2008；Zhao 等人 2010）。在目标 3 中， 来自乙醇暴露的猕猴（Grant U01-INlA-Stress）的样本将被测序。来自 CeA 和皮质区域 25 和 32 的数据将与特定目标 1 和 2 中获得的结果进行比较。