2013-01-012024-05-17https://scholars.lib.ntu.edu.tw/handle/123456789/680331摘要：間腦中的韁核(HA)神經元以及其與中腦腳間核(IPN)的聯結構成了脊椎動物中最保守的神經迴路之一。韁核-腳間核神經迴路功能的重要性包括調控睡眠、回餽及壓迫感等神經生理反應,而且韁核- 腳間核神經迴路的損傷在一些研究上發現與憂鬱症和精神分裂症有關連。此外,在動物模式研究中發現 到藥物濫用造成的主要損傷正是位在韁核-腳間核這個神經迴路上。然而儘管我們在了解韁核-腳間核神 經迴路的解剖圖譜及生理功能上有許多進展,究竟是那些分子和機制控制著韁核-腳間核神經迴路的形成 和功能執行仍然是相當的不清楚。 斑馬魚逐漸演變成研究脊椎動物神經迴路發育及功能主要的模式動物之一,因為在斑馬魚中實驗觀 察、追蹤及遺傳或手術操控神經細胞及迴路的發育過程上要相對的容易。尤其值得注意的是,由於分子 及細胞層次的實驗操控較容易運用在斑馬魚上,因此位在斑馬魚上視丘的松果複合體及韁核近年來被廣 泛利用為一模式來研究腦半球對稱性的生成機制。除此之外,實驗上利用斑馬魚做大規模的正向遺傳篩 選及小分子藥物篩選不但可行且研究結果已使我們在脊椎動物的神經發育和病理生成上有了更深入的 瞭解。 基因體序列的逐漸解碼已被證明在分離及瞭解單ㄧ蛋白質的功能和所參與之細胞生化途徑上為強 有力的工具。然而必需要先瞭解神經迴路內和其間的蛋白質和代謝產物是如何在時間和空間上來協調的 運作才能夠真正的明白神經迴路在生物中是如何的運作。因此為了闡明執行韁核-腳間核神經迴路的參與 分子,首先必需要透過瞭解全腦中與韁核-腳間核神經迴路生理功能有密切關聯的蛋白質體與代謝質體。 在這個研究計劃中,利用斑馬魚作為脊椎動物模式,我的目標是運用遺傳方法學的正向遺傳篩選來 瞭解調控韁核-腳間核神經迴路發育的因子及其操控機制(目標1);以及運用系統生物學方法如蛋白質體學 及代謝質體學等分析策略來發掘探討全腦神經系統中參與執行韁核-腳間核神經迴路功能的分子(目標2 和3)。我預期我的研究將為韁核-腳間核神經迴路的發育與其透過那些因子來發揮功用提供新的見解。我 相信這個研究計畫的結果將會為因生理功能不正常的韁核-腳間核神經迴路所引起的人類疾病其形成原 因提供瞭解的知識基礎。<br> Abstract: The diencephalic habenula nuclei (HA) and their connections to the midbrain interpeduncular nucleus (IPN) forming one of the most highly conserved neural circuit in the vertebrate brain. The importance of HA-IPN circuit has been demonstrated by prior studies showing that the functions of HA-IPN circuit include modulating sleep, reward and stress responses, and lesions in the HA-IPN circuit are linked to a variety of behavior and neurological disorders such as depression and schizophrenia. In addition, major damages of HA-IPN tracks were found in drug-abused animal model. However, despite the considerable progresses in understating the anatomy and physiological function of HA-IPN circuitry, our understanding of the molecules and their operating mechanisms that control the development and function of HA-IPN circuit are largely unknown. Zebrafish has been emerging as one of the major model organism to study the development and function of vertebrate neural circuits because development of neurons and their connections are easier to be visualized, traced and genetically or surgically manipulated. Notably, recent years zebrafish pineal complex and HA in epithalamus have become the major models to study the patterning of hemispherical asymmetry because of their excellent accessibility to cellular and molecular manipulation. In addition, the high-throughput methodologies such as forward genetic and small molecule screens are applicable and have already brought profound insights into our understanding in the development and pathogenesis of vertebrate nerves system. The deciphering of genome sequences has been proved to be powerful in isolating and characterizing functions of individual proteins and the biochemical or cellular pathways they involves. However, to really understand biological processes of neural circuits, we need to understand how functional effectors, the proteins and metabolites, are temporally and spatially coordinated within and around the activated neural circuits. In order to elucidate the functional effectors that execute HA-IPN circuit activities, it is necessary to understand the proteome and metabolome that is closely tied to the physiological functions of HA-IPN circuit inside the whole vertebrate brains. In this research proposal, my goals are to utilize genetic approaches such as forward genetic screens to understand the factors and mechanisms that control HA-IPN circuit development (aim 1), and to utilize system biology approaches such as to proteomic and metabolomic analyses to demonstrate the functional players of HA-IPN circuit at the whole brain level (aim 2 and 3) using zebrafish as the vertebrate model. I expect to bring new insight into our understanding of how the HA-IPN circuit is formed and is executing its functions. I believe the results gained from this project will provide the knowledge bases for understanding the mechanisms underlying the human disorders that are associated with altered physiological function of HA-IPN circuit.韁核-腳間核神經迴路系統生物學遺傳篩選斑馬魚habenula-interpeduncular circuitsystem biologygenetic screenzebrafish