Analysis of miRNA Function During Eye Development and Retinal Regeneration

描述（由申请人提供）：在过去的几年中已经发现了许多非编码RNA，其中最好的特征是被称为微小RNA（miRNA）的21-23个核苷酸的RNA家族。脊椎动物基因组编码数百种miRNA，主要通过抑制翻译在转录后水平调节基因表达。尽管在阐明执行miRNA功能的加工途径和效应复合物方面取得了进展，但对miRNA控制的确切靶mRNA知之甚少。我们一直使用斑马鱼作为模型系统来识别miRNA靶点。在这里，我们建议使用微阵列来识别在斑马鱼视网膜再生过程中差异表达的miRNA，然后功能性测试此类miRNA的参与，以识别控制视网膜再生的关键基因和调节级联反应。 公共卫生相关性：对于人类来说，寿命的延长将增加患有一种或多种眼疾的患者数量。目前，失明或视力受损影响超过300万美国人，但预计到2020年，这一数字将增加到超过500万（Archives of Opthalmology，April，2004）（Ferris III and Tielsch，2004）。许多人类眼睛的疾病涉及光感受器细胞的损失、损伤或退化，但与鱼类相反，再生大多是不存在的。这项提案旨在了解视网膜再生的机制，希望了解允许这种再生的信号可能会确定治疗目标，以恢复或保护光感受器。

DESCRIPTION (provided by applicant): A number of non-coding RNAs have been discovered in the last few years, the best characterized of which are a family of 21-23 nucleotide RNAs referred to as microRNAs (miRNAs). Vertebrate genomes encode hundreds of miRNAs that regulate gene expression at the post-transcriptional level, primarily through repression of translation. Despite progress in elucidating the processing pathways and effector complexes that carry out miRNA function, precious little is known about the exact target mRNAs controlled by miRNAs. We have been using zebrafish as a model system to identify miRNA targets. Here, we propose to perform microarrays to identify miRNAs that are differentially expressed during zebrafish retinal regeneration and then functionally test the involvement of such miRNAs to identify key genes and regulatory cascades that control retinal regeneration. PUBLIC HEALTH RELEVANCE: For humans, increased life span will increase the numbers of patients suffering from one or more eye diseases. Currently, blindness or impaired vision affects over 3 million Americans but that number is expected to increase to over 5 million just by 2020 (Archives of Opthalmology, April, 2004) (Ferris III and Tielsch, 2004). Many diseases of the human eye involve loss, damage or degeneration of photoreceptor cells but in contrast to fish, regeneration is mostly absent. This proposal seeks to understand the mechanisms of retinal regeneration in the hopes that understanding the signals that allow such regeneration might identify therapeutic targets to restore or preserve photoreceptors.