碩士 / 國立臺南大學 / 生物科技學系碩士班 / 102 / The human Leucine-rich Repeats and WD repeat Domain containing 1 (LRWD1) was a high expression gene in the testicular tissues .But the role of LRDW1 function is not clear in the male germ cell . In the past study , the LRWD1 genes was identified by cDNA microarray as one of the down-regulated genes in the testicular tissues of patients with severe spermatogenic defects . The LRWD1 expression was suspected of regulating sperm morphology and motility. Therefore, the purpose of this study is identified LRWD1 functions in vivo. We used shRNA to knockdown LRWD1 expression in NT2D1 and Hela cells. Then ,the degree of cytoskeleton was analysis by Immunofluorescence assay. When the LRWD1 was depleted, microtubule network was destroyed. In order to understand the mechanism of LRWD1, we use zebrafish (Danio rerio) as an animal model to study LRWD1 function . Frist, we understood the expression of LRWD1 at 4~8hpf (hours past fertilization) in zebrafish embryos ,and it maybe had relationship with embryogenesis . In mature zebrafish ,LRWD1 was mainly expressed in testis . In order to further understand LRWD1 function in zebrafish embryos , we used Morpholino oligo(MO) which could block LRWD1 translation . When embryos MO-treated grew to 48hpf , approximately 20% embryo tails were bent . It is interesting that we want to understand LRWD1 function in zebrafish embryogenesis and testis. We performed the constructions of LRWD1 overexpression and knockout zebrafish by gateway system and TALEN . We hope it will be a model organism for drug screening in the future.
| Identifer | oai:union.ndltd.org:TW/102NTNT0111007 |
| Date | January 2014 |
| Creators | Chi-Ping Tai, 戴季平 |
| Contributors | Yen-Ni Teng, 鄧燕妮 |
| Source Sets | National Digital Library of Theses and Dissertations in Taiwan |
| Language | zh-TW |
| Detected Language | English |
| Type | 學位論文 ; thesis |
| Format | 93 |
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