碩士 / 國立臺灣海洋大學 / 生物科技研究所 / 100 / Previously, we cloned and characterized the zebrafish agr2 gene. Whole-mount in situ hybridization demonstrated that zebrafish agr2 is expressed in most organs that contain mucus-secreting cells, including epidermis, olfactory bulbs, otic vesicles, pharynx, esophagus, pneumatic duct, swim bladder, and intestine. However, detailed regulatory mechanisms regarding agr2 expression in specific organ such as otic vesicles are lacking. In this study, by serial deletion, we found -2.5 k to -2.6 k agr2 5’-upstream region can drive EGFP specific expression in otic vesicle of [100ecto-EGFP]- injected embryos. Two SOX10 binding sites were predicted within 100bp DNA sequence between -2.5 k to -2.6 k region using Genomatix, Jaspar and TESS. No EGFP expression can be detected in the otic vesicles of embryos that had been injected with respective recombinant plasmid containing either both mutated SOX10 binding motifs or respective one mutated binding motif. These results indicated that SOX10 binding sites located between-2.5 k to -2.6 k agr2 5’-upstream region are important to direct EGFP expression in the otic vesicles. Moreover, no EGFP expression can be detected in the otic vesicles by using SOX10 MO to knockdown SOX10 in Tg(agr2 FL: EGFP), so we presume SOX10 can regulate agr2 specific expression in otic vesicles. On the other hand, we identified that EGFP is expressed in the semicircular canal and supporting cells of all sensory patches in otic vesicles of Tg(agr2 FL:EGFP) transgenic fish embryos. To understand agr2 gene function in otic vesicles development, we used agr2 MO to test otic vesicles phenotype and inner ear function analysis such as balance, motor coordination and swim bladder inflation but no affect. agr2 expressed in supporting cells, we interested in hair cells regeneration. Therefore, we treated neomycin to induce hair cells death and observe hair cells regeneration. The data showed that agr2 morphant hair cells regeneration function is became weak. We found that agr2 played an important role in supporting cells to regulate hair cells regeneration function.
| Identifer | oai:union.ndltd.org:TW/100NTOU5613007 |
| Date | January 2012 |
| Creators | Chih-Hao Tang, 湯之皓 |
| Contributors | Sheng-Ping L. Hwang, 黃聲蘋 |
| Source Sets | National Digital Library of Theses and Dissertations in Taiwan |
| Language | zh-TW |
| Detected Language | English |
| Type | 學位論文 ; thesis |
| Format | 82 |
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