QTL AND MICROARRAY MAPPING LEAD SENSITIVITY GENES

QTL 和微阵列定位先导敏感性基因

• 批准号: 7117019
• 负责人: Douglas M Ruden
• 金额: $ 32.19万
• 依托单位: WAYNE STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-09-24 至 2009-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7117019
• 关键词: Drosophilidae; RNA interference; behavior disorders; behavioral /social science research tag; biotechnology; developmental neurobiology; environmental exposure; functional /structural genomics; gene expression; genetically modified animals; immunocytochemistry; invertebrate locomotion; lead poisoning; learning disorders; linkage mapping; microarray technology; neuromuscular disorder; neuromuscular junction; polymerase chain reaction; quantitative trait loci; sex behavior; statistics /biometry; synapses

DESCRIPTION (provided by applicant): Human activity has resulted in the environmental distribution of many toxic substances, among them the heavy metals that are spread throughout our biosphere. In addition to the acute toxic effects to humans exposed to lead (i.e. in lead paint or in contaminated drinking water) there are more insidious effects of chronic exposure on the development of all organisms. Children exposed to low levels of lead have altered developmental processes, and these children develop symptoms such as hyperactivity, changes in sensory function, and changes in cognitive abilities ("IQ"). Drosophila is a promising model organism to study the effects of lead exposure during development because of (1) the sophisticated understanding of its genetics, and the ease of manipulating its genome; (2) the availability of behavioral and morphological assays sensitive to small effects of very low doses of lead. There is thought to be a great deal of variability in the sensitivity of lead exposure, and both human and Drosophila cells are thought to induce expression of "protective genes" upon exposure to lead. The hypothesis for this proposal is that one can identify some of the "protective genes" that make an organism resistant to the behavioral and developmental effects of lead toxicity using quantitative trait loci (QTL) mapping techniques combined with microarray and sophisticated genetic analyses. To test this hypothesis, Aim 1 is to map lead sensitive QTL by exposing larvae of 90 recombinant inbred Drosophila strains to environmentally relevant doses of lead, and assaying several behavioral and synaptic connection aspects that are affected by lead - such as larval locomotion, adult learning, adult locomotion, adult locomotion combined with ethanol vapor exposure, and the structure of the larval neuromuscular junction. Aim 2 is to perform DNA microarray analyses with labeled mRNA from whole larvae and adult heads from the recombinant inbred lines that show the greatest differences in lead-induced behavioral alterations, as determined in Aim 1. Aim 3 is to up-regulate, by conditional over-expression, or down-regulate, by using existing mutations and RNA interference techniques, the genes that overlap in the assays performed in Aims 1 and 2, and to determine the effects of lead on cognition, locomotion and synaptic function of these genetically altered flies. Results of these studies will identify candidates for the most important genes that are altered during lead exposure in humans, and could well lead to bioassays or treatments for heavy metal exposure at or below NOAEL and LOAEL values.

描述（由申请人提供）：人类活动导致了许多有毒物质的环境分布，其中包括在整个生物圈中散布的重金属。除了对暴露于铅的人类（即在铅涂料或受污染的饮用水中）的急性毒性作用外，长期暴露对所有生物的发展还有更多阴险的影响。暴露于铅含量低的儿童的发育过程改变了，这些儿童会出现症状，例如多动症，感觉功能的变化以及认知能力的变化（“ IQ”）。果蝇是一种有前途的模型生物体，可以研究发育过程中铅暴露的影响，因为（1）对其遗传学的复杂理解以及操纵其基因组的便利性； （2）行为和形态学测定对极低剂量铅的小作用敏感的。人们认为，铅暴露的敏感性有很大的可变性，人们认为人和果蝇细胞在暴露铅时会诱导“保护基因”的表达。该提议的假设是，可以使用定量性状基因座（QTL）映射技术与微阵列和复杂的遗传分析相结合的一些“保护性基因”，使生物具有抗铅毒性的行为和发育效果。为了检验该假设，目标1是通过揭示90个重组植物果蝇菌株的幼虫来绘制铅的幼虫，以与环境相关的铅剂量，并测定几个受铅影响的行为和突触连接方面，例如铅，例如幼虫，成人学习，成人的成年人段落，成人的成人量表，成人量与成人的范围，成人量落级，远处群体的组合量，远处旋转量，神经肌肉连接。 AIM 2是通过从AIM 1中确定的铅诱导的行为变化的重组亲系中的整个幼虫和成年头的标记mRNA进行DNA微阵列分析，这些杂交线条显示出铅诱导的行为变化的最大差异。目标3是通过有条件地过表达和下调的，或下调了现有的突变和RNA相互作用的概念，从而逐渐划分了概括，从而逐渐划分了概括，从而估算了概括的概括，并逐渐划分了基础，并将其划分为基础，这些构成的概念是，构成了概括的构成，并将其划分为基础。 2，并确定铅对这些遗传改变的苍蝇的认知，运动和突触功能的影响。这些研究的结果将确定人类铅暴露期间最重要的基因的候选者，并且很可能导致在Noael和Loael值以下或低于Noael和Loael值以下的重金属暴露的生物测定或治疗方法。