Please use this identifier to cite or link to this item: http://hdl.handle.net/11455/20216
DC FieldValueLanguage
dc.contributor鄭旭辰zh_TW
dc.contributorHsu-Chen Chengen_US
dc.contributor.author林家貝zh_TW
dc.contributor.authorLin, Chia-Peien_US
dc.contributor.other生命科學系所zh_TW
dc.date2013en_US
dc.date.accessioned2014-06-06T07:12:39Z-
dc.date.available2014-06-06T07:12:39Z-
dc.identifierU0005-3008201311465500en_US
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Hedgehog acts as a somatic stem cell factor in the Drosophila ovary. Nature 410, 599-604, doi:10.1038/35069099 (2001). 35 Ren, Y., Cowan, R. G., Harman, R. M. & Quirk, S. M. Dominant activation of the hedgehog signaling pathway in the ovary alters theca development and prevents ovulation. Mol Endocrinol 23, 711-723, doi:10.1210/me.2008-0391 (2009). 36 Migone, F. F. et al. Dominant activation of the hedgehog signaling pathway alters development of the female reproductive tract. Genesis 50, 28-40, doi:10.1002/dvg.20786 (2012). 37 Dwyer, J. R. et al. Oxysterols are novel activators of the hedgehog signaling pathway in pluripotent mesenchymal cells. J Biol Chem 282, 8959-8968, doi:10.1074/jbc.M611741200 (2007). 38 Kim, S. K. & Melton, D. A. Pancreas development is promoted by cyclopamine, a hedgehog signaling inhibitor. Proc Natl Acad Sci U S A 95, 13036-13041 (1998). 39 Engel, B. D. et al. 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dc.identifier.urihttp://hdl.handle.net/11455/20216-
dc.description.abstract在脊椎動物中,許多器官的發育皆受Hedgehog pathway所調控,而 Hedgehog的成員包括Dhh、Shh和Ihh。以缺失DHH的老鼠為例,無法發育成熟的精子因而導致不育;但雌性依然可以發育成熟的卵子。雌性的生殖細胞在胚胎時期即受RA誘導,開始進行減數分裂;而雄性則是進入發身期後才開始進行減數分裂。在實驗室成員過去的研究中發現,雞的Shh取代Dhh參與精子的發育,對成熟公雞給予外源性Shh後,將使同一截面中精子的形成週期同步化;若給予Cyclopamine則生殖細胞無法繼續發育。為進一步探討Shh在精子形成早期如何進行調控,我們對進入發身期前的公雞,提供RCAS-cShh或20(S)-Hydroxycholesterol,表現PTCH的細胞數目要到第八週才會提升;給予Ciliobrevin A可減少表現PTCH的細胞數目。但無論Hh的活性高低,表現NANOG的細胞數目不受Hh活性影響。顯示生殖幹細胞的數目並不會受Hh活性所影響。而在CAM culture的雌性胚胎卵巢中,給予20(S)-Hydroxycholesterol後表現PTCH的細胞數目增加,且有大量生殖細胞進入減數分裂;給予Cyclopamine後表現TCH的細胞數目減少,而生殖細胞的數目也會減少。因此,在第八週前活化Hh pathway並不會增加PTCH的表現量;生殖幹細胞的數目在發身期前不受到Hh之活性影響;抑制卵巢中Hh之活性將使卵子數目減少。zh_TW
dc.description.abstractIn vertebrate, Hedgehog pathway regulates the development of many organs, there are three members of the HH family: Dhh,Shh and Ihh. The Dhh-null mice are sterility in male owing to absence mature sperm but fertility in female. RA induces germ cells entry of meiosis in female embryonic stage, but not until puberty in male. Our past studies showed that Shh instead of Dhh regulates the development of testis in male chicken. In mature male chicken, ectopic Hh caused synchronous spermatogenesis progression but repressed spermatogenesis by Cyclopamine. To further understand how Hh pathway regulates spermatogenesis, we elevated HH pathway activity in prepubertal testis by providing RCAS-cShh / 20(S)-Hydroxycholesterol( Hh agonist). We found ectopic activating Hh pathway did NOT change the percentage of PTCH+ and NANOG+ cells in W7 testis, but 20(S)-Hydroxycholesterol increased the percentage of PTCH+ cells in W8 testis. Ciliobrevin A decreasing the percentage of PTCH+ cells but not NANOG+ cells. In cultured embryonic ovary, activateing Hh pathway increased the number of PTCH+ germ cells; while inhibiting Hh pathway not only inhibited Ptch expression, but also reduced germ cell number. Hence, ectopic Hh activation could NOT induce precocious spermatogenesis before W8 in testis. The spermatogonia stem cell number is NOT affected by either activation or inhibition of Hh pathway in prepubertal testis. Inhibition of Hh led to fewer germ cells in embryonic ovary.en_US
dc.description.tableofcontents誌謝 ……………………………………………………………………… i 摘要 ……………………………………………………………………… ii Abstract ………………………………………………………………… iii 目次 ……………………………………………………………………… 3 圖目次 …………………………………………………………………… 6 壹、 前言 …………………………………………………………… 9 一、 雄性鳥類之生殖系統發育 ……………………………… 9 二、 雌性鳥類之生殖系統發育 ……………………………… 11 三、 配子進行減數分裂 ……………………………………… 12 四、 RA在生殖系統中所扮演的角色 ………………………… 13 五、 Hedgehog pathway在生殖系統中所扮演的角色 ……… 14 甲、 Hh pathway …………………………………………… 14 乙、 Hh pathway在雄性生殖系統中的調控 …………… 15 丙、 Hh pathway在雌性生殖系統中的調控 …………… 17 貳、 研究目的及試驗假說 ………………………………………… 19 參、 材料與方法 …………………………………………………… 21 一、 試驗動物和受精蛋 ……………………………………… 21 二、 自體羽毛細胞的培養及移植 …………………………… 21 甲、 羽毛細胞培養系統…………………………………… 21 乙、 細胞培養液的配製 ………………………………… 22 丙、 細胞移植手術 ……………………………………… 22 三、 RCAS-cShh之構築與製備………………………………… 22 甲、 構築RCAS-cShh …………………………………… 23 乙、 轉染 ………………………………………………… 23 丙、 病毒液之收集與濃縮 ……………………………… 24 丁、 病毒力價的測定 …………………………………… 24 戊、 功能性測試 ………………………………………… 25 四、 注射藥劑改變公雞睪丸中Hh pathway之活性 ………… 25 五、 性腺培養系統的建立 (CAM culture) …………………… 25 甲、 建立性腺培養系統…………………………………… 26 乙、 添加藥劑改變Hh pathway之活性 ………………… 26 六、 組織固定與石蠟包埋 …………………………………… 26 七、 切片 ……………………………………………………… 27 八、 免疫螢光染色 …………………………………………… 27 九、 量化細胞數目……………………………………………… 28 肆、 試驗結果 ……………………………………………………… 30 一、 分析未進入發身期的公雞睪丸中Hh與Ptch的分佈 …… 30 二、 分析公雞睪丸中Hh、Ptch與NANOG的分佈 ……………… 三、 未進入發身期的公雞之試驗設計流程 ………………… 30 31 甲、 以RCAS-cShh活化Hh pathway ………………… 31 乙、 以20(S)-Hydroxycholesterol活化Hh pathway… 31 丙、 以Ciliobrevin A抑制Hh pathway ……………… 31 四、 調控未進入發身期的公雞睪丸之Hh pathway活性 …… 32 甲、 以RCAS-cShh活化Hh pathway對睪丸發育的影響 ……………………………………………… 32 1. 構築RCAS-cShh ………………………………… 32 2. 測試RCAS-cShh之功能性 …………………… 33 3. RCAS-cShh對發身期前的公雞睪丸之影響…… 33 乙、 以20(S)-Hydroxycholesterol活化Hh pathway對精子發育的影響 ……………………………… 35 丙、 以Ciliobrevin A抑制Hh pathway對精子發育的影響 ………………………………………… 36 四、 觀察胚胎時期卵子的發育情形 ………………………… 形 ……………………………… 38 甲、 建立CAM culture培養性腺 …………………… 38 乙、 添加藥劑改變Hh pathway之活性 …………… 39 1. 以20(S)-Hydroxycholesterol活化Hh pathway對卵子發育的影響 ……………………………… 39 2. 以Cyclopamine抑制Hh pathway對卵子發育的影響 …………………………………………… 40 伍、 討論 …………………………………………………………… 41 一、 分析注射Ciliobrevin A後睪丸中PTCH、HH、NANOG的螢光強度 ………………………………………………… 41 二、 在不同時間點活化睪丸中Hh pathway之比較 ………… 42 三、 在不同時間點抑制睪丸中Hh pathway之比較 ………… 43 陸、 結論 …………………………………………………………… 43 柒、 參考文獻……………………………………………………… 99zh_TW
dc.language.isozh_TWen_US
dc.publisher生命科學系所zh_TW
dc.relation.urihttp://www.airitilibrary.com/Publication/alDetailedMesh1?DocID=U0005-3008201311465500en_US
dc.subjectzh_TW
dc.subjectHedgehogen_US
dc.subject配子zh_TW
dc.subject睪丸zh_TW
dc.subject卵巢zh_TW
dc.subjectgameteen_US
dc.subjecttestisen_US
dc.subjectovaryen_US
dc.titleHedgehog pathway在雞的配子發育中所扮演的角色zh_TW
dc.titleThe role of Hedgehog pathway in chicken gametogenesisen_US
dc.typeThesis and Dissertationzh_TW
item.fulltextno fulltext-
item.languageiso639-1zh_TW-
item.openairetypeThesis and Dissertation-
item.cerifentitytypePublications-
item.openairecristypehttp://purl.org/coar/resource_type/c_18cf-
item.grantfulltextnone-
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