2016-08-012024-05-17https://scholars.lib.ntu.edu.tw/handle/123456789/677872摘要：在胚發育過程中，單細胞受精卵經卵裂分化後成為一成熟個體，但在此期間細胞增生及分化間之調合 機制大多尚未明嘹。在胚胎早期形態改變過程中，胚體會形成不同區間，獨立調控增生及分化，以形 成不同形態。其中中腦背側是發育最快的腦組織，也最適合拿來研究細胞增生及分化之調合機制。我 們之前的研究曾發現一群可使細胞微管不穩定的蛋白基因stathmin，他們在發育早期的大腦中依時間 之不同各有特定表現位置，顯示其可能對斑馬魚腦部發育各自扮演某種角色。我們初步分析其功能發 現其中的stathmin-like 4 (stmn4)可能可抑制神經前驅細胞在中腦之分化，因此本計畫將探討所有 stathmin 在腦發育之角色及其調控機制。我們假設去除斑馬魚之stmn4 會影響中腦神經分化，我們將 嘗試系統性解析所觀察到之表徵跟stmn4 之調控機制。本計畫為三年，其特定目標如下：我們將（一） 針對不同stathmin 在發育中之斑馬魚腦提供高解析度的原位雜合核糖核酸反應及蛋白質組織免疫染色 影像，以更清楚地識別各種不同stathmin 的表現位置。（二）針對不同stathmin 探討其在發育中之斑馬 魚腦神經發育過程之功能。（三）探討stathmin 調節腦神經發育過程之機制。（四）嘗試探討stathmin 與其他已知調節因子如Wnt-FGF-BMP 訊息傳遞路徑之交互作用，以對腦神經發育過程之調節有整<br> Abstract: From the onset of embryonic development, single cell divides and transforms into differentiating cell lineages to make up a mature organism. However, during these sophisticating events, the coordination between proliferation and differentiation remains largely unknown. Compartmentalization of proliferation occurs in early morphogenesis that seems to be critical for patterning. Dorsal midbrain, one of the fastest growing tissues, stands as an attractive platform to understand these phenomena. Previously, we have discovered that stathmin genes, known for their microtubule destabilizing activity, have distinct temporal and spatial expression patterns in the zebrafish developing brain. Our preliminary results further revealed that one of the stathmins, stathmin-like 4 (stmn4), plays a decisive role for preventing neural progenitor cells from differentiation in dorsal midbrain. The goal of this proposal is to reveal the roles of stathmins, in particular stathmin 4 in zebrafish brain development. I hypothesize that the depletion of zebrafish stmn4 will interfere with neurogenesis in dorsal midbrain and plan to systematically analyze these phenotypes and investigate the regulatory mechanism of Stmn4 therein. Specific aims are outlined and to be completed in three years. Specific Aim 1: To provide higher resolution gene expression patterns and protein distributions of zebrafish stathmins in the developing zebrafish brain. Specific Aim 2: To examine the functional roles of stathmins on neurogenesis in the developing brain. Specific Aim 3: To investigate underlying mechanisms of how stathmins regulate neurogenesis in the developing brain. Specific Aim 4: To understand the regulatory network between stathmins and Wnt-FGF-BMP signaling for the modulation of brain neurogenesis. Collectively our results will demonstrate stathmin-like 4 might work as a crucial role to guarantee the neural progenitor cells in dorsal midbrain against premature differentiation, leading to malformation of tectum. Roles of other stathmins in the developing brain will also be elucidated in zebrafish.stathmin4/RB3HuC/elavl3微管G2 期Wntstathmin4/RB3HuC/elavl3microtubuleG2 phaseWnt 表