GENETICS OF DEVELOPMENTAL PLASTICITY IN THE ADIPOCYTE

脂肪细胞发育可塑性的遗传学

• 批准号: 6167215
• 负责人: Leslie Paul Kozak
• 金额: $ 41.96万
• 依托单位: LSU PENNINGTON BIOMEDICAL RESEARCH CTR
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-09-01 至 2005-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6167215
• 关键词: adipocytes; artificial chromosomes; brown fat; cell differentiation; cytogenetics; developmental genetics; fluorescent dye /probe; gene expression; gene induction /repression; gene interaction; genetic mapping; genetic regulation; genetically modified animals; genotype; laboratory mouse; molecular cloning; nucleic acid sequence; polymerase chain reaction; quantitative trait loci; sequence tagged sites; serial analysis of gene expression; tissue /cell culture; transcription factor

DESCRIPTION (Adapted from investigator's abstract): This is a new RO1 application from a senior investigator that requests 5 years of support to identify genes that control the induction of brown adipocytes in white fat tissue. Prior work, much of it based on seminal contributions of the PI, indicates that brown fat thermogenesis mediated by Uncoupling protein 1 can have a significant impact on energy homeostasis, and that activation of this program can proceed through previously existing brown fat but also, importantly, by the development of new brown fat within white adipose tissue of adult animals. Using Ucp1 mRNA levels as a marker for the extent of brown fat induction in retroperitoneal fat (which is normally white fat and so does not express Ucp1), the PI proposes to further characterize and identify 4 or 5 so-called Iba genes that have been identified in preliminary QTL and RI studies of B6 vs. A/J mice. Each of the QTLs has been localized to a 10 - 15 cM interval; in Aim 1, Iba genetic architecture will be investigated by constructing B6.A and A.B6 congenic lines for each of the loci, then measuring the effect of those QTLs by themselves and in 16 different combinations. For QTLs whose effect remains robust when examined in isolation as a congenic line, further genetic crosses will be carried out in Aim 2 to narrow the region; in addition, corresponding strains from a genome-wide panel of B6.CAST congenics will be evaluated for the Iba phenotype and, if positive, used in parallel with the B6 vs. A/J cross. In Aim 3, BAC contigs will be generated for loci whose position can be narrowed to 1 - 2 cM (the PI suggests that Iba4 on chromosome 19 meets this criteria), and candidate genes will be identified by direct sequencing and reference to B6 and A/J retroperitoneal fat cDNA libraries the PI proposes to construct. In parallel (Aim 4), SAGE analysis will be undertaken using retroperitoneal fat RNA from the congenic lines. Finally (Aim 5), candidate cDNAs or genes will be tested by complementation in a cell culture system based on preadipocytes or BAC transgenesis using B6.A congenics.

描述（改编自研究者的摘要）：这是一个新的 RO1 高级研究员的申请，要求 5 年的支持 鉴定控制白色脂肪中棕色脂肪细胞诱导的基因 组织。之前的工作，其中大部分基于 PI 的开创性贡献， 表明解偶联蛋白 1 介导的棕色脂肪产热作用可以 对能量稳态有重大影响，并且这种激活 程序可以通过先前存在的棕色脂肪进行，而且， 重要的是，通过在白色脂肪组织内形成新的棕色脂肪 成年动物。使用 Ucp1 mRNA 水平作为棕色脂肪范围的标记 腹膜后脂肪（通常是白色脂肪，因此不 表达 Ucp1)，PI 建议进一步表征和识别 4 或 5 在初步 QTL 和 RI 研究中已鉴定出的所谓 Iba 基因 B6 与 A/J 小鼠的比较。每个 QTL 均定位于 10 - 15 cM 间隔;在目标 1 中，将通过以下方式研究 Iba 遗传结构： 为每个基因座构建 B6.A 和 A.B6 同系系，然后测量 这些 QTL 本身以及 16 种不同组合的效果。为了 当作为同类系进行单独检查时，其效果仍然强劲的 QTL， 目标2将进行进一步的基因杂交以缩小区域；在 此外，来自 B6.CAST 同源基因全基因组组的相应菌株 将评估 Iba 表型，如果呈阳性，则与 B6 与 A/J 交叉。在目标 3 中，将为以下基因座生成 BAC 重叠群： 位置可以缩小到 1 - 2 cM（PI 表明染色体上的 Iba4 19符合此标准），候选基因将通过直接鉴定 B6 和 A/J 腹膜后脂肪 cDNA 文库的测序和参考 PI建议建设。同时（目标 4），将进行 SAGE 分析 使用来自同类系的腹膜后脂肪RNA。最后（目标 5）， 候选 cDNA 或基因将通过细胞培养物中的互补进行测试 基于前脂肪细胞或使用 B6.A 同源基因的 BAC 转基因的系统。