Novel approach to select genes from RMA normalized microarray data using functional hearing tests in aging mice
Novel approach to select genes from RMA normalized microarray data using functional hearing tests in aging mice
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DOI：
10.1016/j.jneumeth.2008.02.022
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发表时间：
2008-06-30
影响因子：
3
通讯作者：
Frisina, Robert D.
中科院分区：
文献类型：
--
作者：
D'Souza, Mary;Zhu, Xiaoxia;Frisina, Robert D.
老年性耳聋-与年龄有关的听力损失-是头号沟通障碍和我们老年人口的三大慢性疾病之一。高通量技术有可能用于鉴定差异表达的基因，这些基因可能是感觉和神经疾病更好的诊断和治疗靶点。在这里，我们分析了一组GABA受体的基因表达在老化CBA小鼠的耳蜗使用Affyssin基因芯片MOE 430 A。功能表型听力的措施，包括听觉脑干反应（ABR）阈值和畸变产物耳声发射（DPOAE）振幅（四个年龄组）。使用四个特定标准来评估来自RMA标准化微阵列数据的基因表达变化（40个重复）。使用线性回归模型将神经生理学听力测量值拟合到探针集表达谱。这些数据首先进行单因素方差分析，然后进行线性回归。此外，对数信号比转换为倍数变化，并通过相对实时PCR确认选定的基因表达变化。主要发现：GABA-A受体亚基α 6的表达随着年龄和听力损失而上调，而亚基α 1则受到抑制。此外，GABA-A受体相关蛋白样-1和GABA-A受体相关蛋白样-2随着年龄和听力损伤而强烈下调。最后，使用Pathway Architect将基因表达测量与内耳相关的途径/网络关系相关联，以确定与观察到的基因表达变化一致的关键途径。(c)2008 Elsevier B. V.保留所有权利。
Presbycusis - age-related hearing loss - is the number one communicative disorder and one of the top three chronic medical condition of our aged population. High-throughput technologies potentially can be used to identify differentially expressed genes that may be better diagnostic and therapeutic targets for sensory and neural disorders. Here we analyzed gene expression for a set of GABA receptors in the cochlea of aging CBA mice using the Affymetrix GeneChip MOE430A. Functional phenotypic hearing measures were made, including auditory brainstem response (ABR) thresholds and distortion-product otoacoustic emission (DPOAE) amplitudes (four age groups). Four specific criteria were used to assess gene expression changes from RMA normalized microarray data (40 replicates). Linear regression models were used to fit the neurophysiological hearing measurements to probe-set expression profiles. These data were first subjected to one-way ANOVA, and then linear regression was performed. In addition, the log signal ratio was converted to fold change, and selected gene expression changes were confirmed by relative real-time PCR Major findings: expression of GABA-A receptor subunit alpha 6 was upregulated with age and hearing loss, whereas subunit alpha 1 was repressed. In addition, GABA-A receptor associated protein like-1 and GABA-A receptor associated protein like-2 were strongly downregulated with age and hearing impairment. Lastly, gene expression measures were correlated with pathway/network relationships relevant to the inner ear using Pathway Architect, to identify key pathways consistent with the gene expression changes observed. (c) 2008 Elsevier B.V. All rights reserved.