碩士 / 國立臺灣大學 / 漁業科學研究所 / 92 / Type I (acidic) and type II (basic) keratins, two major classes of intermediate filament proteins, express mainly in epithelial cells in mammals. However, they are little known in zebrafish. Using a partial cDNA sequence of a zebrafish type I keratin zkrt18-like as probes to observe the mRNA expression pattern of zkrt18-like, the zkrt18-like mRNA signal was detected in one cell stage embryos by whole mount in situ hybridization, suggesting that the zkrt18-like mRNA was inherited through the maternal effect. The zkrt18-like expressed in envelope layer of 8 hours-post-fertilization (hpf) embryos, and it expressed in all surface cells after 10 hpf. Moreover, the zkrt18-like transcripts not only presented in epidermal cells but also presented in the venter of 20 hpf embryos. Through the cross section of 48 hpf embryos, the expression in venter was observed to be in the dorsal aorta, gut and pronephric duct. Comparing the expression profile of the reporter gene and the endogenetic zkrt18-like mRNA by an established transgenic line zkrt18-like-RFP, which contained the upstream 3.3 kb of zkrt18-like fused with red fluorescent protein (rfp), the RFP expression in all the progeny from transgenic F1 female mated with wild-type male in one cell stage suggests that the rfp regulated by zkrt18-like promoter was also inherited through the maternal effect. The RFP expression of the F2 from transgenic F1 male mated with wild-type female (zkrt18-like-RFP♂ F2) was on the surface of 14 hpf embryos, and kept expressing in epidermal cells. Through the cross section of 48 hpf zkrt18-like-RFP embryos, the RFP expression was also found in dorsal aorta, gut and pronephric duct, indicating that the RFP expression was similar to the zkrt18-like expression pattern. Detecting the rfp mRNA of 8 hpf and 10 hpf zkrt18-like-RFP♂ F2 embryos by whole mount in situ hybridization, it has been demonstrated that the rfp transcripts was in accordance with the endogenous zkrt18-like mRNA expression. By RT-PCR analysis, the rfp expression of zkrt18-like-RFP♂ F2 initiated at 3 hpf, suggesting that the zkrt18-like expression initiated during the mid-blastula stage and conformed the mid-blastula transition phenomenon in fish and amphibian embryos. Besides, through the observation on the zkrt18-like-RFP larva and adult, the ZKRT18-LIKE was observed to exist in eyes, intestine, liver, scale epidermis, and spinal cord during its whole life. This result is different from those keratins in mammals, which mainly express in epithelial cell.
| Identifer | oai:union.ndltd.org:TW/092NTU05451001 |
| Date | January 2004 |
| Creators | Yu-Ju Lin, 林育如 |
| Contributors | Huai-Jen Tsai, 蔡懷楨 |
| Source Sets | National Digital Library of Theses and Dissertations in Taiwan |
| Language | zh-TW |
| Detected Language | English |
| Type | 學位論文 ; thesis |
| Format | 58 |
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