IDENTIFICATION OF THE MOLECULAR MECHANISMS OF ZEBRAFISH INNER EAR HAIR CELL REGENERATION USING HIGHTHROUGHPUT GENE EXPRESSION PROFILING

All nonmammalian vertebrates studied can regenerate inner ear mechanosensory receptors, i.e. hair cells, but mammals only possess a very limited capacity for regeneration after birth. As a result, mammals suffer from permanent deficiencies in hearing and balance once their inner ear hair cells are lost. The mechanisms of hair cell regeneration are poorly understood. Because the inner ear sensory epithelium is highly conserved in all vertebrates, we chose to study the hair cell regeneration mechanism in adult zebrafish, hoping the results would be transferrable to inducing hair cell regeneration in mammals. We defined the comprehensive network of genes involved in hair cell regeneration in the inner ear of adult zebrafish with the powerful transcriptional profiling technique, Digital Gene Expression (DGE), which leverages the power of next-generation sequencing. We also identified a key pathway, stat3/socs3, and demonstrated its role in promoting hair cell regeneration through stem cell activation, cell division, and differentiation. In addition, transient pharmacological up-regulation of stat3 signaling accelerated hair cell regeneration without over-producing cells. Taking other published datasets into account, we propose that the stat3/socs3 pathway is a key response in all tissue regeneration and thus an important therapeutic target not only for hair cell regeneration, but also for a much broader application in tissue repair and injury healing. The dissertation contains four supplemental files. Supplemental file 1 contains raw data of five expression profiles generated by DGE. It is a tab-delimited text file with six columns. The first column contains the sequences of the tags and the second to sixth columns contain the count of the corresponding tags in control, 0-hpe, 24-hpe, 48-hpe, and 96-hpe profiles respectively. Supplemental file 2 contains UniGene clusters identified from unambiguously mapped tags. It is a tab-delimited text file with six columns. The first column contains the UniGene IDs. The second to sixth columns contain the count of the corresponding UniGene clusters in control, 0-hpe, 24-hpe, 48-hpe, and 96-hpe profiles respectively. Supplemental file 3 contains candidate genes identified by comparison of the expression profiles during regeneration to the control profiles. It is a tab-delimited text file with 19 columns. The contents in each column are specified in the header. Supplemental file 4 contains a list of the candidate genes known to be expressed in the inner ear and/or the lateral line system during development. It is a tab-delimited text file with four columns which contain UniGene IDs, ZFIN IDs, Entrez Gene IDs, and gene symbols respectively