Gene expression and function of autocrine motility factor receptor (amfr) during zebrafish development

• Main Authors: Jia-Yi Deng, 鄧嘉宜
• Other Authors: Wen-Wen Lin
• Format: Others
• Language: zh-TW
• Published: 2009
• Online Access: http://ndltd.ncl.edu.tw/handle/13097022384742744104

碩士 === 國立高雄海洋科技大學 === 海洋生物技術研究所 === 97 === Autocrine motility factor receptor (AMFR) is a membrane glycoprotein receptor that plays an important role in stimulating tumor cell motility, inducing angiogenesis, and inhibiting apoptosis. Clinically, AMFR is one of the biomarkers for prognostic evaluation. AMFR is an ubiquitin protein ligase (E3) involved in endoplasmic reticulum associated degradation (ERAD). Molecular cloning and sequencing have revealed that AMF is the same as a glucose phosphate isomerase (GPI), neuroleukin (NLK) or maturation factor (MF), is a multifunctional protein. Though the cellular activities of AMFR have been the focus in the previous studies, the function of AMFR during embryonic development remains unclear. We aimed to study the gene expression of zebrafish amfr and function of gpi/ amf during embryonic development. Publicly available genomic sequences reveal that zebrafish amfr is located at chromosome 25 that spans about 20 kb containing 13 exons. Northern blot hybridization shows that 4.2 kb is the main transcript expressed in vaious tissues and developmental stages. The results of RT-PCR and southern blot hybridization also confirm that amfr is expressed in many tissues and developmental stages. Whole-mount in situ hybridization shows that amfr mRNA is maternally deposited in the cleavage and blastula embryos and zygotic expression is detected in yolk syncytial nuclei at late blustula, gastrulation and segmentation stages. During pharyngula to hatching stages, amfr is specifically expressed in brain, eyes, liver, gut, pectoral fin, and muscle. Microinjection of gpia/ gpib siRNA results in morphants of incomplete epiboly, growth delay, heart malformation, pericardial edema, and bent tail. These phenotypes can be rescued by co-injection of gpia/ gpib capped RNA. Our results suggest that gpia/ gpib functions in the process of epiboly, providing metabolic energy, and development of muscle and heart.