碩士 / 國立臺灣大學 / 生化科技學系 / 99 / Maternal-to-zygotic transition (MZT) plays an important role in early embryogenesis. In this study, zebrafish embryos at stages of 256-cell and sphere were used as the standard samples for pre- and post-MZT. Totally, 17 proteins expressed differentially during MZT were picked out by two-dimensional electrophoresis (2-DE). Six of these proteins were identified as hypothetical protein LOC768289, mitochondrial processing peptidase alpha (MPP-alpha), aldehyde dehydrogenase 2b (ALDH2b), methylmalonate-semialdehyde dehydrogenase (MMSDH) and ubiquitin C-terminal hydrolase L1 (UCHL1) by using ESI-QUAD-TOF. These proteins might relate to mitochondrial protein import pathway, branched-chain amino acid metabolism, or cell cycle regulation. Furthermore, we detected the mRNA expression of MPP-alpha, ALDH2b, MMSDH, UCHL1 and LDH (lactate dehydrogenase) by semi quantitative RT-PCR. These results were comparable to 2-DE data. The expression differences of UCHL1, MPP-alpha and LDH during MZT were statistically significant. In an attempt to study the function of these proteins during MZT, we used the specific inhibitors, oxamate and LDN57 444, to inhibit LDH or UCHL1, respectively. The phenotypes of cleavage delay and dorsalis were observed in inhibitors treated embryos. Additionally, we found that LDH activity in 256-cell was higher than that in sphere by LDH activity assay. In conclusion, our results demonstrated that the change of the protein profile during MZT may relate to mitochondrial energy metabolism. These proteins can be further studied by using specific monoclonal antibody, transgenic animal model, or RNA interference.
| Identifer | oai:union.ndltd.org:TW/099NTU05106113 |
| Date | January 2011 |
| Creators | Hua-Shan Shih, 施驊珊 |
| Contributors | 莊榮輝 |
| Source Sets | National Digital Library of Theses and Dissertations in Taiwan |
| Language | zh-TW |
| Detected Language | English |
| Type | 學位論文 ; thesis |
| Format | 66 |
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