碩士 / 國立高雄海洋科技大學 / 海洋生物技術研究所 / 100 / Autocrine motility factor receptor (AMFR) is a glycoprotein originally named gp78. AMFR/gp78 plays a dual role as a cytokine receptor on the cell surface or as an E3 ubiqui¬tin ligase involved in endoplasmic reticulum associated degradation (ERAD). AMFR has important clinical implication, which has prognostic value in clinical cancer research. However, the exact function of AMFR in embryo¬nic development remains un¬clear. Using in situ hybridization to detect amfr mRNA in zebrafish embryos, we found that amfr mRNA was localized in blastomere and yolk syncytial layer (YSL) at mid-blastula stage. The amfr expression patten is correlated with the development of YSL from mid-blastula to late gastrulation. YSL appears to play an important role in the formation of 3 germ layers, therefore we pro¬pose that amfr is essential to the function of YSL. Antisense morpholino knock¬down of amfr pro¬duced defective phenotypes and caused epibolic delay in zebrafish embryos. Defective phenotypes of amfr knockdown embryos include bent tail, pericar¬dial edema, malformed muscles, cerebral hypoplasia and abnormal yolk cell extension. The percentage of defective phenotypes increased as the dosage of amfr antisense morpholino injection increased. We found that amfr morphant with epib¬oly delay also exhibited abnormalities in mesoendodermal, mesodermal and endo¬dermal cell migration. To study the mechanism of epiboly delay caused by amfr knockdown, we analyzed the effect on microtubule by assessing microtubule and tubulin contents in amfr knockdown em¬bryos. Our analyses indicated that the con¬tent of microtubule increase in embryos treated with microtubule stabiliz¬ing agent taxol and in amfr knockdown embryos. Immunohistochemical analysis also showed that both amfr knockdown and taxol treatment produced dense and bundles of microtubule in yolk cytoplasmic layer. These results suggest that amfr knock¬down inhibits the depolymertization of yolk cell microtubule. We propose that amfr expressed in the YSL functions in the epibolic movement and formation of three germ layers.
| Identifer | oai:union.ndltd.org:TW/099NKIMT270001 |
| Date | January 2011 |
| Creators | Chin - Wei Chang, 張晉維 |
| Contributors | Wen-Wen Lin, 林文文 |
| Source Sets | National Digital Library of Theses and Dissertations in Taiwan |
| Language | zh-TW |
| Detected Language | English |
| Type | 學位論文 ; thesis |
| Format | 69 |
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