Neurocircuitry Mapping and Genotyping Core

神经回路图谱和基因分型核心

基本信息

批准号：
8521480
负责人：
ROBERT J. HITZEMANN
金额：
$ 37.59万
依托单位：
OREGON HEALTH & SCIENCE UNIVERSITY
依托单位国家：
美国
项目类别：
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/8521480
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 sequencing protein function segregation tool transcriptome sequencing

项目摘要

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）：这是对U01赠款的竞争续订申请，标题为“神经循环映射和基因分型核心”； 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年）和b）制定定量RNASEQ的策略（Bottomly等，2011;附录A）。慢性间歇性乙醇（CIE）程序与适当的对照组将由Crabbe U01提供。由于HDID和对照是从8条近交菌株交叉得出的（Hitzemann etal。1994），鉴于掩盖了掩盖寡核苷酸阵列数据的数据，RNASEQ是有用的（参见Walter等，2007,2009）。 n = 32/group;先前的工作（Lancu等人，2010年）表明，这种大小的样本足以适合拟议的分析。样品是通过激光捕获微截断（LCM）收集的；分析的区域优先级将是杏仁核（CEA）> Infralimbic Cortex（IL）>前胞皮层（PL）的中央核。枕皮层（OC）将用作对照区域。 AIM 1专注于暴饮暴食，而AIM 2则重点关注慢性乙醇暴露在有限的2瓶选择范式中如何影响乙醇消耗。我们的工作假设是，共表达网络和单个基因的差异表达之间的差异具有最大的翻译价值（例如，参见Oti等人2008； Zhao等人，2010年）。在AIM 3中，将对乙醇暴露猕猴（Grant U01- iNLA应力）的样品进行测序。将来自CEA和皮质区域25和32的数据与特定目标1和2中获得的结果进行比较。