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博碩士論文 etd-0209109-151158 詳細資訊
Title page for etd-0209109-151158
論文名稱
Title
前列腺素D2合成脢於雌雞腦下垂體前葉細胞經由過氧化脢體增殖因子活化受體路徑誘導黃體激素表現
The cPGDS induces LHB expression in primary culture of hen anterior pituitary cells via the PPAR signaling pathway
系所名稱
Department
畢業學年期
Year, semester
語文別
Language
學位類別
Degree
頁數
Number of pages
67
研究生
Author
指導教授
Advisor
召集委員
Convenor
口試委員
Advisory Committee
口試日期
Date of Exam
2008-12-03
繳交日期
Date of Submission
2009-02-09
關鍵字
Keywords
前列腺素、過氧化脢體增殖因子活化受體、腦下垂體、黃體激素
PPAR, PGDS, LHB, pituitary
統計
Statistics
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中文摘要
前列線素D2合成脢(chicken prostaglandin D2 synthase, cPGDS)為本實驗室先前發現之高產候選基因,於包含單冠白來航雞的多種品系中皆顯示在高產蛋母雞表現量較高。為進一步瞭解其下游分子機制,我們探討了cPGDS對於雞隻初代培養細胞之作用。我們成功的備製了高專一性之cPGDS多價抗體。在十三個組織樣本中,定量反轉錄聚合酶酵素連鎖反應與西方轉漬顯示cPGDS mRNA與蛋白質高量的表現在小腸、腎臟、肝臟與腦下垂體。比較相同週齡雞隻,cPGDS於雌雞腦下垂體及卵巢表現量亦顯著高於雄雞腦下垂體與睪丸。轉染pEGFP-cPGDS至含1 mM花生油四烯酸(arachidonic acids)培養之腦下垂體前葉初代細胞,誘導了貝他黃體激素(luteinizing hormone beta subunit; LHB)的轉錄與轉譯。於腦下垂體前葉初代細胞添加環氧合酶(cyclooxygenase)路徑代謝產物前列線素D2、前列線素J2, 同樣的誘導了LHB之表現且呈現時間及劑量上的相關。添加過氧化酶體增殖因子活化受體(Peroxisome proliferator-activated receptors; PPARs)促進劑亦有相同的現象。此外,經特異性之PPARA與PPARG抑制劑處理,顯著的抑制了PGJ2誘導之LHB表現。總結而論,前列線素合成酶D2於雌雞腦下垂體前葉細胞會經由過氧化酶體增殖因子活化受體路徑誘導黃體激素表現。
Abstract
Our previous study identified one candidate transcript, chicken prostaglandin D2 synthase (cPGDS), significantly higher expressed in hens with high-egg production across several chicken strains including the Single-Comb White Leghorn layers. To further elucidate its underlying molecular mechanisms, the downstream effects of cPGDS in primary culture of chicken anterior pituitary cells were investigated. We successfully generated a highly specific rabbit anti-cPGDS polyclonal antibody. Of 13 examined tissues/cells, quantitative reverse transcription polymerase chain reaction and Western blotting analysis demonstrated that cPGDS mRNA and protein were highly expressed in intestine, kidney, liver and pituitary gland. In addition, significantly higher cPGDS mRNA and protein expression levels of the pituitary gland and ovary in hens than of the pituitary gland and testis in roosters with the same age were found. Transfection of pEGFP-cPGDS plasmid into primary culture of pituitary anterior cells in medium that contained 1 mM arachidonic acids induced luteinizing hormone beta subunit (LHB) transcription and subsequent translation. Treatments of two metabolites in the cyclooxygenase pathway, PGD2 and PGJ2 in the primary culture of chicken anterior pituitary cells induced LHB transcription and translation in both dose- and time-dependent manners. Treatment of peroxisome proliferator activated receptors (PPARs) agonist increased LHB transcription and translation in chicken anterior pituitary cells. On the other hand, specific inhibitions of PPARA and PPARG using antagonists dramatically suppressed PGJ2-induced LHB transcription level. Taken together; these data suggested PGDS upregulated LHB transcription in primary culture of chicken anterior pituitary cells via the PPAR signaling pathway.
目次 Table of Contents
Contents
Abbreviations…………………………………………………………... Ⅰ
Abstract
Chinese …………………………………………………………. Ⅱ
English …………………………………………………………. Ⅲ
Literature Review……………………………………………………….. 1
Introduction……………………………………………………………... 10
Materials and methods………………………………………………….. 13
Results…………………………………………………………………… 21
Discussion………………………………………………………………... 27
Figures and Tables………………………………………………………. 34
Supplementary Data…………………………………………………….. 49
References………………………………………………………………... 52
Appendix…………………………………………………………………. 60
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