碩士 / 淡江大學 / 生命科學研究所碩士班 / 98 / Abstract:
PART i :
von Hippel-Lindau Disease is a rare condition associated with familial cancers. It is due to a mutation of the VHL tumour suppressor gene at chromosome site 11q13, 3p26-p25. VHL can degrade hypoxia-inducible factors (HIF1a, HIF2a) in the presence of oxygen, thus regulating a lot of hypoxia responsive genes indirectly. pVHL forms a complex with elongins B and C, Cullin2, Rbx1, thus regulating RNA polymerase II and inactivating transcription. After sequence comparison, we found that the zebrafish VHL polypeptide shares sequence identities of 43%, 40%, 45%, 45%, 43% and 45% with the reported VHL of Xenopus, chicken, human, dog, , mouse and mice, respectively. Renal cell carcinoma is the leading cause of death in patients with von Hippel-Lindau disease, with a prevalence as high as 76% reported in one autopsy series. Here, we used zebrafish as an animal model to study it''s roles during early kideny developmental stage. Using morpholino knockdown approach, Our data showed that knock VHL down led to defect of pronephric duct, pronephric tubule and glomerulus. We also find that angiogenesis becoming more strongly after morpholino knockdown. Taken together, we concluded that VHL is required for zebrafish kidney and vascular development.
PART ii:
ATP2A family genes encode all kinds of the SERCA Ca2+-ATPases, which are intracellular pumps located in the sarcoplasmic or endoplasmic reticula of muscle cells. Those enzymes catalyze the hydrolysis of ATP coupled with the translocation of calcium from the cytosol to the sarcoplasmic reticulum lumen, and is involved in muscular excitation and contraction. We used zebrafish as an animal model to study their roles during early developmental stage. After sequence comparison, we found that the zebrafish ATP2A1(ATP2A2a) polypeptide shares sequence identities of 87%(87%), 88%(88%), 87%(87%), 87%(87%), 87%(87%) and 87%(87%) with the reported ATP2A1 (ATP2A2a) of Xenopus, chicken, human, dog, , mouse and mice, respectively. Whole mount in situ hybridization experiments showed that ATP2A1 was first detected at 1cell embryos, and was expressed filled the entire animal pole of 6 hours post-fertilization (hpf) and 12hpf embryos. At the later stage, ATP2A1 was expressed at 18-somite of 18hpf, and was expressed at trunk muscle cells of 24hpf and 31hpf embryos. ATP2A1 was expressed at eye muscle cells, jaw muscle cells, and trunk muscle cells after 48hpf embryos. We also found that ATP2A1 was expressed at Intestinal after 72hpf embryos. Whole mount in situ hybridization experiments also showed that ATP2A2a was first detected at 1cell embryos, and was expressed at mesoderm on both sides of the embryo of 6hpf, 12hpf and 18hpf embryos and was expressed at cardiac precursors of 18hpf embryos. At the later stage, ATP2A2a was expressed at cardiac muscle cells and trunk muscle cells of 24hpf, 31hpf and 48hpf. ATP2A1 was expressed at eye muscle cells, jaw muscle cells, cardiac muscle cells and trunk muscle cells after 72hpf, but the Signal in the trunk muscle cells becomed more weak.
| Identifer | oai:union.ndltd.org:TW/098TKU05105010 |
| Date | January 2010 |
| Creators | Yen-Yu Lai, 賴彥佑 |
| Contributors | Yau-Hung Chen, 陳曜鴻 |
| Source Sets | National Digital Library of Theses and Dissertations in Taiwan |
| Language | zh-TW |
| Detected Language | English |
| Type | 學位論文 ; thesis |
| Format | 97 |
Page generated in 0.0082 seconds