DIFFERENTIAL GENE EXPRESSION IN NORMAL, MUTANT AND AGED

正常、突变和老年的差异基因表达

• 批准号: 6379604
• 负责人: MARGARET I. LOMAX
• 金额: $ 21.74万
• 依托单位: UNIVERSITY OF MICHIGAN AT ANN ARBOR
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-09-27 至 2003-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6379604
• 关键词: aging; cochlea; congenital deafness; developmental genetics; gene expression; laboratory mouse; microarray technology; mutant; neurogenetics; northern blottings; polymerase chain reaction; sensorineural hearing loss

DESCRIPTION (Applicant's abstract): The overall scientific objectives of this proposal are to identify genes whose expression changes as a consequence of two types of sensorineural deafness in the mouse: age-related hearing loss in strain C57BL6/J, and congenital deafness in three deafness mouse mutants. These mutants are being studied at the University of Michigan by two of the co-investigators on this grant. Shaker-2 has been shown to carry mutations in the gene for an unconventional myosin, MyoXV, by Dr. Camper's group. Pirouette and spinner mutants have been mapped and the mutated genes are being identified by Dr. Kohrman's group. We propose to determine the broad effects on the transcriptional repertoire (transcriptome) of the inner ear due to single gene mutations that result in deafness in the mouse. This approach is based on two related hypotheses: (1) these mutations alter the normal developmental processes and homeostatic mechanisms that operate during maturation of the cochlea or in the adult organism, and (2), the alterations will be reflected in changes in the steady state transcript levels of genes that operate in the relevant developmental and homeostatic pathways. By using highly parallel methods to identify gene expression changes, we will begin to characterize the regulatory circuits that are affected directly or indirectly by the mutations, and which themselves may also play critical roles in normal inner ear development, homeostasis and function. Aims 1 and 2 propose to use currently available reagents and technique, such as gene arrays on nylon membranes, to examine gene expression changes in C57BL/6J, a mouse model of age-related hearing loss (Aim 1), and in the three mouse models of congenital deafness and vestibular dysfunction (Aim 2). The final two Aims address the feasibility of applying new and emerging techniques to increase the 'completeness' of the pool of profiled inner ear genes (Aim 3) through database comparisons and SAGE analysis, and the 'throughput' of expression profiling by developing DNA microarrays on glass slides containing genes expressed in the mouse inner ear (Aim 4). These studies should enhance our understanding of the molecular events of age-related hearing loss and congenital deafness and provide new reagents for assessing changes in gene expression in the ear.

描述（申请人摘要）：本研究的总体科学目标 一个建议是确定基因的表达变化的结果，两个 小鼠感觉神经性耳聋的类型： C57 BL 6/J株和三种耳聋突变小鼠的先天性耳聋。这些 密歇根大学的两名研究人员正在研究突变体。 共同研究者Shaker-2已被证明携带突变， 一种非传统的肌球蛋白基因MyoXV，由Camper博士的团队开发。后肢回旋 spinner突变体已经被绘制出来，突变基因正在被鉴定出来， 科尔曼博士的团队我们建议确定对 内耳的转录库（转录组）由于单基因 导致小鼠耳聋的突变。这种方法基于两个 相关假设：（1）这些突变改变了正常的发育 过程和稳态机制，在成熟的运作过程中， 耳蜗或成年生物体，和（2），改变将反映在 基因的稳态转录水平的变化，这些基因在细胞中起作用， 相关的发育和稳态途径。通过使用高度并行 鉴定基因表达变化的方法，我们将开始表征 直接或间接受突变影响的调节回路， 它们本身也可能在正常的内耳中发挥关键作用 发育、体内平衡和功能。目标1和2建议目前使用 可用的试剂和技术，如尼龙膜上的基因阵列， 检查C57 BL/6 J中的基因表达变化，C57 BL/6 J是一种与年龄相关的小鼠模型， 听力损失（目标1），并在三个小鼠模型的先天性耳聋和 前庭功能障碍（目的2）。最后两个目标涉及以下可行性： 应用新的和新兴的技术，以增加'完整性'的池 通过数据库比较和SAGE， 分析，以及通过开发DNA表达谱的“通量”， 载玻片上的微阵列含有小鼠内耳中表达的基因 (Aim 4）。这些研究应该能增强我们对分子事件的理解 与年龄相关的听力损失和先天性耳聋，并提供新的试剂 用于评估耳朵中基因表达的变化。