2017-10-012024-05-14https://scholars.lib.ntu.edu.tw/handle/123456789/659722摘要：粒粒線體是調節細胞生理功能之重要樞紐，近年的研究顯示，粒線體產生能量與伴隨而來的活性氧化物質在許多疾病中扮演關鍵的角色。雌性生殖系統中，粒線體數量與其DNA的完整性亦與濾泡品質、卵巢老化有關。其中粒性細胞在濾泡中可扮演調節內分泌或支持卵母細胞之角色；而本團隊先前的研究已發現，由粒性細胞分化而成的大黃體細胞中，粒線體數目可隨著動情周期的進程而增加。然而在濾泡時期中，相關之結構變化仍待進一步的研究加以分析。 粒線體透過分裂、融合、生合成與粒線體自噬等機制維持其數量與品質。本研究即希望透過山羊卵巢組織超微結構之分析，尋找探討粒線體動態變化的最佳時機，輔以高通量基因表現分析與細胞實驗來驗證其機制。研究可分為三大主題： 1.監測山羊動情周期中濾泡之發育，精確採樣供三維超微結構分析。 2.使用高通量基因表現分析，尋找調控粒線體動態變化之機制。 3.在細胞模式中使用過氧化&#37238;報導基因，探討各基因在細探討各基因在細胞中的分布，並透過質譜蛋白質體學之分析，驗證活體之機制。 本研究在生理狀態下，藉由探討粒線體結構變化之機制，可望對於提升繁殖效率、治療不孕症與其他相關疾病有所幫助。<br> Abstract: Mitochondria serve as the pivot point in cell physiology. Researches have shown that the generation of energy (ATP) and reactive oxygen species (ROS) from mitochondria impact on the cellular responses in diseases. In female reproductive systems, the mitochondrial quantity and mitochondrial DNA integrity have been linked to the quality of follicles and the ovarian aging. As the granulosa cells can regulate the endocrine functions or support the developments of oocytes in follicles, they also differentiate into large luteal cells (LLC) in corpus luteum. Our previous studies have demonstrated that mitochondria are increased in LLC during the progresses of estrous cycle, while the changes in follicles remain to be studied. The quality and quantity of mitochondria can be controlled by the dynamic events including fission, fusion, biogenesis and mitophagy. Based on ultrastructural analysis, present study is planned to search the optimized timing for high throughput gene expression studies. The regulatory mechanisms will be further confirmed in cell model. The research can be divided into three parts: 1.Monitor the developments of the follicles for accuracy sampling and 3D ultrastructural analysis 2.High throughput gene expression analysis for the mechanisms regulating mitochondrial dynamics. 3.With the peroxidase reporter genes in cell model, study the 3D distribution of target genes and conduct the subcellular proteomes by mass spectrum for verifications. Present study will investigate the mechanisms regulating mitochondrial structural changes under physiological conditions. The knowledge to be gained will have the potential to improve the efficiency of reproductions and help the treatments for infertility.山羊黃體濾泡電子顯微鏡超微結構粒線體次世代定序蛋白質體學報導基因goatcorpus luteumfollicleelectron microscopyultrastructuremitochondrianext generation sequencingproteomicsreporter genes