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Please use this identifier to cite or link to this item: http://ntour.ntou.edu.tw:8080/ir/handle/987654321/48568

Title: 點帶石斑魚利用甲基睪固酮誘導之早熟雄魚期性別不穩定之因素探討
Immediately Sex Reversal in the 17α-methyltestosterone-Induced Female Orange-Spotted Grouper, Epinephelus coioides
Authors: Andri Iskandar
Contributors: NTOU:Department of Aquaculture
Keywords: 性轉變;點帶石斑魚;甲基睪固酮;芳香環轉化酶抑制劑
sex-change;orange-spotted grouper;MT;AI
Date: 2015
Issue Date: 2018-08-22T06:06:13Z
Abstract: 先雌後雄的點帶石斑為台灣重要的養殖經濟魚類,生殖腺為精卵混合型,然而我們對於石斑之性轉變過程仍尚未清楚,因此本研究想了解投餵雄性素(MT)之石斑幼魚,停餵後的性逆轉現象,是否會受到甲基化抑制物(5-Aza)影響其生殖體細胞之變化。第一個實驗中,以MT投餵並同時注射5-Aza於石斑,控制組的皆為雌魚,MT處理組的皆為雄魚,MT後停餵組的皆再次性逆轉為雌性,此結果表明利用投餵MT使石斑魚性轉變的方法是不穩定的,而在注射5-Aza的雌性石斑魚並沒有性轉變成雄性。此外,由MT投餵的石斑魚生長速度較慢,顯示長期投餵MT對生長可能有副作用。第二個實驗中,與生殖細胞分化發育相關之dmc1、nanos2及sycp3在停餵AI和MT組的樣本中並無趨勢,這顯示上述基因可能無法有效地區分在混和性生殖腺中雌雄生殖細胞的個別變化。
The present study was expected to scrutinize the instrument of sex-change in fish in the early change process, whether the testis that is turned around from immature ovary using 17α-methyltestoterone (MT) would recover after the termination of MT treatment. We also connected MT-induced sex-change and 5-aza-2’-deoxycytidine (5-Aza) as DNA methylation inhibitor to comprehend the alternation of gonadal soma cells. In the first experiment, we used the orange-spotted groupers at the developmental ages and fed a diet containing MT at 50 mg/kg for 3 months and then a normal diet for 1 month. Intraperitoneally injection of 5-Aza was examined in the first week and second week during the MT-oral administration. Most of the fishes in the control group had immature ovaries, but all the females fed with MT, had an immature spermatogenesis. However, one month after the withdrawal of MT treatment, the sex of fish was returned to femaleness even the fish have undergone MT-induced masculinization. This outcome demonstrates precocious sex-change from underyearling orange-spotted grouper by means of oral MT treatment is impermanent. All the female of 5-aza treatments showed no spermatogenic cells. In this study, lower growth rates were demonstrated by the MT treated groups. The impact of this metabolic change was clear after the end of the hormone oral administration since the decreased growth of the groups treated for 3 months. The second experiment demonstrated the dmc1, nanos2 and sycp3 gene expression related to gonadal development to consider the cell-sexual re-programming of the cells in the AI/MT-terminated male-to-female sex-changed fish. Juvenile female fish were fed a diet containing AI/MT for 3 months and then AI/MT fed was terminated. To analyze the proliferating activities and cell tracing in the fish, we used BrdU injection at a different time. To obtain the gene expression of the fish gonad at different developmental days, we used q-PCR analysis. Our result revealed that during AI-induced sex-change, dmc1 genes showed significantly lower expression compared to the control fish in 0 d, 2 weeks and 8 weeks. The expression levels of dmc1 genes were increased in the MT-treated fish, and the differences were significant after 8 weeks of termination compared to the control fish. The higher expression of nanos2 genes showed at 4 weeks after termination in the MT treated fish
URI: http://ethesys.lib.ntou.edu.tw/cgi-bin/gs32/gsweb.cgi?o=dstdcdr&s=G0010333044.id
Appears in Collections:[水產養殖學系] 博碩士論文

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