摘要：Tristetraprolin (TTP)是一具有鋅手指的RNA結合蛋白，可結合在mRNA的3’端未轉譯區域之ARE（AU-rich element）上誘發去腺甘酸的作用而造成mRNA的快速衰變，當細胞受到外來刺激如生長因子、胰島素、或脂多醣等會激發TTP的快速表現，再作用於帶有ARE的mRNA上如一些前致癌基因、細胞素、及生長因子等，以後轉錄調控方式控制這些基因的表現。最近發現TTP可在老鼠卵巢中卵泡發育的特定時期表現，目前已知這中間有許多轉錄因子參與其中，但關於卵子發育時的後轉錄調控尚未有清楚的研究。本計劃即是要探究TTP在卵子成熟與卵泡發育時所扮演的角色。首先我們觀察其在這些過程中的RNA和蛋白質表現；另外我們也想要知道TTP在卵子發育時的標的RNAs，抽取有TTP表現的卵巢細胞質萃取液，以TTP的抗體進行免疫沉澱，將被沉澱下、與TTP結合的RNA分離出，再反轉錄成DNA後進行cDNA矩陣雜交，可得知TTP的作用標的，再進一步分析探究TTP與標的RNAs在生化和功能上的交互作用。我們的研究將有助於對卵子形成、發育和成熟過程中的分子調節機轉有更清楚的了解。
Abstract: Tristetraprolin (TTP) is a zinc-finger containing RNA binding protein. TTP binds AU-rich elements (AREs) of target mRNAs such as proto-oncogenes, cytokines and growth factors, and then induces deadenylation or directs them to the exosome or associates with RISC-microRNA complexes for rapid degradation of target mRNAs. It was observed as an immediate-early gene that was induced in response to several kinds of stimuli, such as insulin and other growth factors and stimulators of innate immunity like lipopolysaccharides (LPS). Recently, TTP was shown that expressed in mouse ovary. However, little is known about its functional significance during folliculogenesis and oogenesis. In this project, we are to explore the expression pattern and function role of TTP in oocyte development. At first, we will demonstrate its mRNA and protein expression during oogenesis and follicle development by using real-time PCR , in situ hybridization, Western blotting and immunohistochemistry analyses. Then, the TTP targeted mRNAs will be identified to investigate the detailed molecular mechanism of TTP-mediated ovarian development. The immunoprecipitation will be performed by anti-TTP antibody to bring down the mRNA subsets bound to TTP. The TTP-associated mRNAs are to be isolated and reverse-transcriptased to DNA and followed by cDNA array hybridization. Further functional and biochemical assays will be carried out to characterize the interaction between TTP and its target mRNAs. This study will help us understand the TTP-mediated posttranscriptional regulation in oocyte formation and maturation and provide a possible aspect to control these processes.