臺灣博碩士論文加值系統

光是控制許多鳥類生理過程之重要外部因素，季節性繁殖鳥類睪丸的形態與功能會經歷動態季節性變化，睪丸活動之變動決定公鵝每年繁殖週期之變動，光波長模式亦為影響家禽生產性能之重要因子，已有許多研究探討不同光波長模式對生長性狀及雌性繁殖性狀之影響，目前尚缺乏不同光波長模式對雄性繁殖性狀效應之探討。本研究之目的為探討季節變化與光波長對白羅曼鵝睪丸蛋白質表現之影響。試驗一共使用9隻3歲齡白羅曼公鵝，分別於非繁殖季時期(7月)、性腺恢復期(12月)與繁殖時期(2月)採血分析賀爾蒙與犧牲採集睪丸供蛋白質分析；結果顯示繁殖時期之公鵝睪丸重量及睪固酮濃度顯著高於非繁殖期與性腺恢復期(P < 0.05)，蛋白質二維差異電泳分析發現有124個蛋白質點於不同繁殖時期具差異表現，應用質譜分析可成功鑑定其中107個點之身分，其分屬於74個不同蛋白質。基因功能分析結果顯示此等差異表現蛋白質主要位於細胞質(20%)及細胞膜(16%)，主要與蛋白質結合(24%)與催化活性(16%)等分子功能相關，並參與生物性調節(24%)及細胞過程(24%)等生物過程。試驗二共使用30隻3歲齡白羅曼公鵝，分別接受30勒克斯之紅光、白光與藍光處理，鵝隻於試驗起始點、試驗第6週、試驗第20週與試驗第30週採血供賀爾蒙分析與犧牲採集睪丸供蛋白質分析，鵝隻並於試驗第8週起每兩週採集精液供精液性狀分析。結果顯示試驗第30週時白光處理組公鵝之血中睪固酮與雌二醇濃度比值高於其他兩組者(P < 0.05)；藍光處理組公鵝精液之精子活力、存活率與正常形態精子百分比皆低於白光處理者(P < 0.05)。蛋白質二維差異電泳分析發現有140個蛋白質點於不同光波長處理間有差異表現，質譜分析成功鑑定其中122個蛋白質點之身分，且分屬86個不同蛋白質。基因功能分析結果顯示此等差異表現蛋白質主要位於細胞質(26%)及細胞膜(19%)，主要參與蛋白質結合(23%)與碳水化合物結合(18%)等分子功能有關，且大多與細胞過程(28%)與生物性調節(23%)等生物過程有關。綜而言之，公鵝在不同繁殖期之睪固酮濃度與睪丸蛋白質表現不同，不同光波長會影響睪丸蛋白質表現，且藍光處理會降低公鵝精子活力、存活率與正常精子百分比，然其對繁殖力影響之確切機制有待進一步研究。

Light is an important exogenous factor that controls various physiological processes in birds. Testes of birds undergo dynamic changes of their morphology and function in various to seasonal variations. The fluctuations in testicular activity determine the transition between stages of the annual reproductive cycle. Patterns of light wavelength are important factors that influence productive performances in modern poultry industry. Previously studies showed that different patterns of light wavelength affected growth performances and reproductive traits in females. There is still no study on the effects of patterns of light wavelength on male reproductive traits. The purposes of this study were to investigate protein expression in the testes of ganders in different breeding stages and whether different light wavelengths affect the protein expression in the testes of ganders. In the first experiment, 9 three-years-old male White Roman ganders were used. The ganders were sacrificed at non-breeding stage in July, sexual reactivation stage in December, and breeding stage in February to collect blood and testis samples for hormone and proteomic analysis, respectively. The results showed that the testicular mass and level of testosterone of ganders in the breeding stage were greater than those in non-breeding and sexual reactivation stages (P < 0.05). Two-dimensional difference gel electrophoresis analysis showed that there were 124 protein spots differentially expressed in the testes of ganders at different reproductive stages. A total of 107 protein spots were identified by mass spectrometry and belonged to 74 different proteins. Gene ontology analysis revealed that most of the differentially expressed proteins located in cytoplasm (20%) and membrane (16%). Most of the proteins belonged to molecular function of protein binding (24%) and catalytic activity (16%) and participated in biological process of biological regulation (24%) and cellular process (24%). The second experiment used 30 three-year-old White Roman ganders. The ganders were treated with white, red, and blue light of 30 lux intensity. The semen quality traits were evaluated every two weeks after 8 weeks of light treatment. The ganders were sacrificed at the beginning, sixth, twentieth, thirtieth weeks of treatment to collect blood and testis samples for hormone and proteomic analysis, respectively. The results showed that the testosterone to estradiol ratio in the white light treated ganders was high than those in the other groups at thirtieth weeks of light treatment (P < 0.05). Sperm motility, viability and percentage of sperm with normal morphology of ganders in blue light group were lower than those in the white light treated group (P < 0.05). Two-dimensional difference gel electrophoresis revealed that there were 140 protein spots differentially expressed in the testes of ganders under different light wavelength treatments. A total of 122 spots were identified by mass spectrometry and belonged to 86 unique proteins. Gene ontology analysis revealed that the differentially expressed proteins were majorly located in cytoplasm (26%) and membrane (19%), belonged to molecular function of protein binding (23%) and carbohydrate binding (18%), and participated in biological process of cellular process (28%) and biological regulation (23%). In conclusion, ganders at different reproductive stages showed different levels of testosterone and expression of proteins in testes. Different light wavelength affected the protein expression in the testes of ganders. Blue light treatment reduced the sperm motility, viability and percentage of sperm with normal morphology, and expression of proteins related to sperm maturation and capacitation. The exact mechanisms of that seasonal change and light wavelength affect fertility of ganders requires further investigation.

中文摘要 i
Abstract ii
目錄 iv
表次 vi
圖次 vii
壹、前言 1
貳、文獻探討 2
一、 鳥禽季節性繁殖 2
(一) 季節性繁殖 2
(二) 亞熱帶與溫帶鳥禽季節性繁殖 2
(三) 熱帶鳥禽季節性繁殖 3
二、 光照對鳥禽繁殖生理之影響 3
(一) 鳥禽光之感知 3
(二) 光影響鳥禽類生理生殖之路徑 5
(三) 光對鳥禽下視丘-腦垂腺-性腺軸與神經內分泌系統之影響 6
(四) 光照期對鳥類生理之影響 8
(五) 光強度對鳥類生理之影響 9
(六) 光波長對鳥類生理之影響 9
三、 鵝之繁殖季節性 11
(一) 鵝隻繁殖特性 11
(二) 光照對鵝隻繁殖生理之影響 12
(三) 公鵝生殖系統介紹 12
四、 蛋白質體與其應用 13
五、 研究目的 14
參、季節變化對白羅曼公鵝睪丸蛋白質表現之影響 15
一、 材料與方法 15
(一) 試驗鵝隻之品種、來源 15
(二) 試驗設計 15
(三) 血清中性腺賀爾蒙濃度之分析 18
(四) 鵝隻睪丸蛋白質體分析 18
(五) 統計分析 21
二、 結果 22
(一) 不同繁殖期公鵝睪丸重量之生長 22
(二) 不同繁殖期公鵝血清中性腺賀爾蒙濃度差異 23
(三) 不同繁殖期公鵝睪丸蛋白質表現 24
(四) 不同繁殖時期公鵝睪丸蛋白質差異表現蛋白質之功能分類 34
三、 討論 35
(一) 不同繁殖期公鵝睪丸生長與性腺賀爾蒙之關係 35
(二) 不同繁殖期公鵝睪丸差異表現蛋白質與睪固酮之關係 35
(三) 不同繁殖期公鵝睪丸差異表現蛋白質之功能探討 36
肆、光波長對白羅曼公鵝睪丸蛋白質表現之影響 39
一、 材料與方法 39
(一) 試驗鵝隻之品種、來源 39
(二) 試驗設計 39
(三) 血清中性腺賀爾蒙濃度之分析 41
(四) 公鵝精液採集與性狀分析 41
(五) 鵝隻睪丸蛋白質體分析與生物資訊分析 41
(六) 統計分析 41
二、 結果 43
(一) 不同光波長對公鵝性腺賀爾蒙之影響 43
(二) 不同光波長對公鵝精液性狀之影響 43
(三) 不同光波長對公鵝睪丸蛋白質表現之影響 46
(四) 不同光波長中公鵝睪丸蛋白質差異表現蛋白質之功能分類 57
三、 討論 58
(一) 不同光波長對公鵝性腺賀爾蒙與公鵝精液性狀之影響 58
(二) 不同光波長對公鵝睪丸蛋白質表現效應之影響 58
伍、結論 60
陸、參考文獻 61
柒、附表 70

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