碩士 / 中山醫學大學 / 生物醫學科學學系碩士班 / 104 / Human TMPRSS3 which is a member of the Type II transmembrane serine protease (TTSP) was the first description of a serine protease involved in deafness. In our previous studies, we have used zebrafish as a model organism to explore tmprss4a and tmprss4b which are the homologue of human TMPRSS3. We also found that zebrafish tmprss3 may be a homologue of human TMPRSS3. The main purpose of this study is to further explore and understand the role of zebrafish tmprss3. We found that both zebrafish tmprss3a and tmprss3-like are probably homologous to human TMPRSS3 and mouse Tmprss3 through BLAST (Basic Local Alignment Search Tool). During embryogenesis, reverse transcription polymerase chain reaction (RT-PCR) revealed that zebrafish tmprss3a was expressed after 1.5 post-fertilization (hpf), and a relatively high level of expression was detected between 25-120 hpf. On the other hand, zebrafish tmprss3-like was expressed from 1.5 to 120 hours hpf, but a relatively low level of expression was detected between 9-12 hpf. In addition, both tmprss3a and tmprss3-like transcripts were observed in multiple adult tissues through RT-PCR. Moreover, the highest expression of tmprss3a were detected in heart, kidney, second is innear. On the other hand, the highest expression of tmprss3-like was in vas deferens, second is kidneys. Whole-mount in situ hybridization revealed that the signals of tmprss3a and tmprss3-like were gradually obvious during embryogenesis for longer. However, the signal of tmprss3a was relatively low in the early stage. tmprss3a and tmprss3-like are specially express in the ventral spine at 24-hour stage. In addition, we developed one specific tmprss3-like-targeting morpholino oligonucleotides (MOs) and established the fusion protein construct TagRFP tmprss3-like that was co-injected into embryos in the presence or absence of morpholinos. We further knocked-down tmprss3-like by MOs, and observed that the application of 4ng MO could decrease the expression of pTaqRFP tmprss3-like fluorescent from 85.67 ± 0.08% to 23 ± 0.06%. In addition, MO against pTagRFP fluorescent do not be affected, so our results may prove that tmprss3-like Mo would be specific knockdown the expression of pTaqRFP tmprss3-like fluorescence. In summary, although we contributes to the understanding of the specific expression and distribution of tmprss3a and tmprss3-like in the embryonic development and tissue, the specific roles of tmprss3a and tmprss3-like in the zebrafish still have be investigate.
| Identifer | oai:union.ndltd.org:TW/104CSMU5114005 |
| Date | January 2016 |
| Creators | Ying-Ru Lee, 李盈儒 |
| Contributors | Jiann-Jou Yang, 楊建洲 |
| Source Sets | National Digital Library of Theses and Dissertations in Taiwan |
| Language | zh-TW |
| Detected Language | English |
| Type | 學位論文 ; thesis |
| Format | 61 |
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