Morris Ken A, Snir Einat, Pompeia Celine, Koroleva Irina V, Kachar Bechara, Hayashizaki Yoshihide, Carninci Piero, Soares M Bento, Beisel Kirk W
Department of Biomedical Sciences, Creighton University, 2500 California Plaza, Omaha, NE 68178, USA.
J Assoc Res Otolaryngol. 2005 Mar;6(1):75-89. doi: 10.1007/s10162-004-5046-x. Epub 2005 Apr 22.
Microarray analyses have contributed greatly to the rapid understanding of functional genomics through the identification of gene networks as well as gene discovery. To facilitate functional genomics of the inner ear, we have developed a mouse inner-ear-pertinent custom microarray chip (CMA-IE1). Nonredundant cDNA clones were obtained from two cDNA library resources: the RIKEN subtracted inner ear set and the NIH organ of Corti library. At least 2000 cDNAs unique to the inner ear were present on the chip. Comparisons were performed to examine the relative expression levels of these unique cDNAs within the organ of Corti, lateral wall, and spiral ganglion. Total RNA samples were obtained from the three cochlear-dissected fractions from adult CF-1 mice. The total RNA was linearly amplified, and a dendrimer-based system was utilized to enhance the hybridization signal. Differentially expressed genes were verified by comparison to known gene expression patterns in the cochlea or by correlation with genes and gene families deduced to be present in the three tissue types. Approximately 22-25% of the genes on the array had significant levels of expression. A number of differentially expressed genes were detected in each tissue fraction. These included genes with known functional roles, hypothetical genes, and various unknown or uncharacterized genes. Four of the differentially expressed genes found in the organ of Corti are linked to deafness loci. None of these are hypothetical or unknown genes.
微阵列分析通过识别基因网络以及发现基因,极大地促进了对功能基因组学的快速理解。为了推动内耳的功能基因组学研究,我们开发了一种与小鼠内耳相关的定制微阵列芯片(CMA-IE1)。从两个cDNA文库资源中获得了非冗余cDNA克隆:日本理化学研究所(RIKEN)的内耳消减文库和美国国立卫生研究院(NIH)的柯蒂氏器文库。芯片上存在至少2000个内耳特有的cDNA。进行比较以检查这些独特cDNA在柯蒂氏器、外侧壁和螺旋神经节中的相对表达水平。从成年CF-1小鼠的三个耳蜗解剖部分中获得总RNA样本。对总RNA进行线性扩增,并利用基于树枝状大分子的系统增强杂交信号。通过与耳蜗中已知的基因表达模式进行比较,或与推断存在于这三种组织类型中的基因和基因家族进行相关性分析,来验证差异表达基因。阵列上约22%-25%的基因有显著的表达水平。在每个组织部分中都检测到了一些差异表达基因。这些基因包括具有已知功能作用的基因、假设基因以及各种未知或未表征的基因。在柯蒂氏器中发现的四个差异表达基因与耳聋位点相关。这些基因均不是假设基因或未知基因。