Animal & Resource Core

项目摘要 此次中心更新包括动物和资源核心(C001)，反映了 研究企业，重点放在共享的遗传小鼠模型和集成 行为表型鉴定和高通量测序服务。为满足这些要求，C001将1) 建立和维护遗传小鼠模型群体；2)执行标准操作程序(SOP) 支持：基于酒精偏好水平的短期选择性育种，通过两个月的长期乙醇消费 瓶子选择(2BC)程序，以及集合转移程序对认知灵活性的评估； 简化核糖核酸/脱氧核糖核酸样本的提交和处理程序，以供 通过C001的测序部实现大规模并行测序共享资源(MPSSR)。集中化 C001中的这些角色和职责将使项目调查员从开发的负担中解放出来 不同的资源和程序集合，从而为解决相关问题提供了更大的自由度 实验操作和执行尖端分析，以解决特定的假设。动物与动物 资源核心计划将在整个中心范围内提供多种优势，包括降低研究成本 通过规模经济，通过标准化提高科学严谨性和实验重复性 畜牧业和蜂群维护实践，以及由专职工作人员在 恒定可控的环境。C001将与研究项目合作完成四个 明确的目标。在目标1中，核心将建立、维护和分发遗传小鼠模型，以支持 研究项目中心。模型是异种股票合作杂交(HS-CC)小鼠， 有选择地培育出高(HP)和低(LP)乙醇偏好的小鼠系，以及ALDH1L1-EGFP-Rpl10a 转基因小鼠。在目标2中，核心将执行行为表型服务，包括那些SOP 以及有助于解释基因组和蛋白质的显著效应的特征 饮酒的药理调控，如味觉偏好、运动活性和酒精 通行证。在目标3中，HS-CC小鼠的集合转换过程将得到优化，这与认知能力密切相关 在猕猴身上进行的测试，从而加强了中心项目之间的跨物种转换。 这一SOP将用于在HP和LP小鼠身上进行P001的研究，以解决认知与 灵活性与酒精摄入量和P001和P004的风险，以解决剂量依赖的后果 长期摄入乙醇。在目标4中，动物和资源核心的测序部将执行下一步- 为所有研究项目提供代测序服务，并将与生物信息学合作 核心(C002)，用于RNA-seq和全基因组dNaM-seq数据的比对和分析。

Project Summary The inclusion of an Animal & Resource Core (C001) in this Center renewal reflects the emergent needs of a research enterprise with an augmented focus on shared genetic mouse models and demands for integrated behavioral phenotyping and high-throughput sequencing services. To address these requirements, C001 will 1) establish and maintain colonies of genetic mouse models; 2) conduct standard operating procedures (SOPs) for: short-term selective breeding based on level of ethanol preference, chronic ethanol consumption via a two- bottle choice (2BC) procedure, and the evaluation of cognitive flexibility with a set-shifting procedure; and 3) provide a streamlined process for the submission and processing of RNA/DNA samples for sequencing by the Massively Parallel Sequencing Shared Resource (MPSSR) via the Sequencing Division of C001. Centralizing these roles and responsibilities within C001 will relieve project investigators from the burden of developing separate sets of resources and procedures, thereby providing greater latitude to address pertinent experimental manipulations and perform cutting-edge analyses to address specific hypotheses. Animal & Resource Core initiatives will offer multiple Center-wide advantages, including reduction of research costs through an economy of scale, enhanced scientific rigor and experimental reproducibility through standardized husbandry and colony maintenance practices, and implementation of optimized SOPs by dedicated staff in a constant and controlled environment. C001 will collaborate with the Research Projects in the completion of four Specific Aims. In Aim 1, the Core will establish, maintain and distribute genetic mouse models to support the Center Research Projects. The models are the Heterogeneous Stock Collaborative Cross (HS-CC) mice, selectively bred mouse lines for high (HP) and low (LP) ethanol preference, and Aldh1l1-EGFP-Rpl10a transgenic mice. In Aim 2, the Core will perform behavioral phenotyping services, including those SOPs mentioned above, but also for traits that aid in the interpretation of significant effects of genomic and pharmacological manipulations on ethanol drinking, such as tastant preference, locomotor activity, and ethanol clearance. In Aim 3, a set-shifting procedure will be optimized in HS-CC mice that closely parallels cognitive testing performed in rhesus macaques, thereby enhancing cross-species translation between Center projects. This SOP will be used to perform studies for P001 in HP and LP mice that address the relationship of cognitive flexibility with risk for ethanol intake and for P001 and P004 to address the dose-dependent consequences of chronic ethanol intake. In Aim 4, the Sequencing Division of the Animal & Resource Core will perform next- generation sequencing services for all Research Projects, and will work in conjunction with the Bioinformatics Core (C002) for the alignment and analysis of RNA-seq and genome-wide DNAm-seq data.