Analysis of Cochlear Protein Profiles of Wistar, Sprague-Dawley, and Fischer 344 Rats with Normal Hearing Function
Analysis of Cochlear Protein Profiles of Wistar, Sprague-Dawley, and Fischer 344 Rats with Normal Hearing Function
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DOI：
10.1021/pr900222c
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发表时间：
2009-07-01
影响因子：
4.4
通讯作者：
Coling, Donald
中科院分区：
文献类型：
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作者：
Jamesdaniel, Samson;Ding, Dalian;Coling, Donald
耳蜗蛋白表达的差异可能会影响不同动物模型对特定类型听觉病理的易感性。然而，目前对内耳中大量表达的蛋白质知之甚少。这些蛋白质的鉴定可能有助于寻找易感性和干预靶点的生物标志物。为了开始解决这个问题，我们分析了耳蜗蛋白质的三个品系的大鼠，Wistar，Sprague-Dawley，和Fischer 344，使用广谱抗体微阵列。采用畸变产物耳声发射（DPOAE）检测动物的正常听觉功能。在整个耳蜗中筛选的725种蛋白质中，超过80%在所有三种菌株中检测到。然而，它们发生的水平有着显著的差异。在213种表达水平为？的蛋白质中：2倍的肌动蛋白，在所有三个菌株中仅检测到7.5%。肌球蛋白轻链激酶（MLCK）免疫定位于外毛细胞（OHC）的表皮板，丝裂原活化蛋白激酶（MAP）-细胞外信号调节激酶1/2（ERK 1/2）免疫定位于OHC、柱细胞、螺旋韧带边缘细胞和成纤维细胞。文献综述表明，这16种蛋白质中有7种（44%）的表达是首次在内耳中检测到的，尽管其中一些蛋白质的存在有一定的意义。这些丰富的，但未经研究的蛋白质，MAP激酶激活的蛋白激酶2（MAPKAPK 2），显示在柱细胞和内毛细胞（IHC）强免疫标记。在OHC、支持细胞、神经元、边缘细胞、中间细胞和基底细胞中有中度MAPKAPK 2标记。目前的研究提供了多个大鼠品系的第一个大的耳蜗蛋白质谱。这些菌株中丰富的蛋白质表达的多样性可能导致这些菌株对衰老、噪音或耳毒性药物的敏感性差异。
Differences in the expression of cochlear proteins are likely to affect the susceptibility of different animal models to specific types of auditory pathology. However, little is currently known about proteins that are abundantly expressed in inner ear. Identification of these proteins may facilitate the search for biomarkers of susceptibility and intervention targets. To begin to address this issue, we analyzed cochlear protein profiles of three strains of rats, Wistar, Sprague-Dawley, and Fischer 344, using a broad spectrum antibody microarray. Normal hearing function of the animals was ascertained using distortion product otoacoustic emissions (DPOAE). Of 725 proteins screened in whole cochlea, more than 80% were detected in all three strains. However, there were striking differences in the levels at which they occur. Among 213 proteins expressed at levels ?:2 fold of actin, only 7.5% were detected at these levels in all three strains. Myosin light chain kinase (MLCK) was immunolocalized in cuticular plate of outer hair cells (OHC) while mitogen activated protein (MAP) kinase-extracellular-signal regulated kinase1/2 (ERK1/2) was detected as foci in OHC, pillar cells, strial marginal cells, and fibroblasts of spiral ligament. A review of literature indicated that the expression of 7 (44%) of these 16 proteins were detected for the first time in the inner ear, although there were implications of the presence of some of these proteins. One of these abundant, but unstudied, proteins, MAP kinase activated protein kinase2 (MAPKAPK2), shows strong immunolabeling in pillar cells and inner hair cells (IHC). There was moderate MAPKAPK2 labeling in OHC, supporting cells, neurons, and marginal, intermediate, and basal cells. The current study provides the first, large cochlear protein profile of multiple rat strains. The diversity in expression of abundant proteins in these strains may contribute to differences in susceptibility of these strains to aging, noise, or ototoxic drugs.