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将基因组学应用于鸟类内耳:开发用于探索感觉功能和毛细胞再生的消减cDNA资源。

Applying genomics to the avian inner ear: development of subtractive cDNA resources for exploring sensory function and hair cell regeneration.

作者信息

Hawkins R David, Helms Cynthia A, Winston Julia B, Warchol Mark E, Lovett Michael

机构信息

Division of Human Genetics, Department of Genetics, Washington University School of Medicine, 4566 Scott Ave., St. Louis, MO 63110, USA.

出版信息

Genomics. 2006 Jun;87(6):801-8. doi: 10.1016/j.ygeno.2005.12.014. Epub 2006 Mar 3.

Abstract

We applied a micro-cDNA-based subtraction method to identify genes expressed in the regenerating sensory epithelia (SE) of the chicken inner ear. Sensory hair cells in the avian utricle SE are in a constant state of turnover, where dying hair cells are replaced by new ones derived from supporting cells. In contrast, hair cells in the cochlea remain quiescent unless damaged. We used this difference to enrich for utricle-specific genes, using reiterative cDNA subtraction and demonstrate enrichment for utricle-specific sequences. A total of 1710 cDNA sequence reads revealed the presence of many cDNAs encoding known structural components of the SE (for example, Harmonin and beta-tectorin), proteins involved in cellular proliferation, such as P311, HIPK2, and SPALT1, among many others of unknown function. These libraries are the first of their kind and should prove useful for the discovery of candidate genes for hearing disorders, regenerative and apoptotic pathways, and novel chicken ESTs.

摘要

我们应用了一种基于微cDNA的消减方法来鉴定鸡内耳再生感觉上皮(SE)中表达的基因。鸟类椭圆囊SE中的感觉毛细胞处于不断更新的状态,死亡的毛细胞被来自支持细胞的新毛细胞取代。相比之下,耳蜗中的毛细胞除非受损,否则保持静止。我们利用这种差异,通过反复的cDNA消减来富集椭圆囊特异性基因,并证明了椭圆囊特异性序列的富集。总共1710个cDNA序列读数显示存在许多编码SE已知结构成分的cDNA(例如,Harmonin和β-耳纤毛蛋白)、参与细胞增殖的蛋白质,如P311、HIPK2和SPALT1,以及许多其他功能未知的蛋白质。这些文库是同类文库中的首批文库,应该对发现听力障碍、再生和凋亡途径的候选基因以及新的鸡ESTs有用。

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