The development of female germ cells in the coral, Euphyllia ancora

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Project title

Code/計畫編號

NSC101-2313-B019-012-MY3

Translated Name/計畫中文名

雌性腎形真葉珊瑚生殖細胞的發育作用探討

Project Coordinator/計畫主持人

Ching-Fong Chang

Funding Organization/主管機關

National Science and Technology Council

Department/Unit

Department of Aquaculture

Website

https://www.grb.gov.tw/search/planDetail?id=2865459

Year

2013

Start date/計畫起

01-08-2013

Expected Completion/計畫迄

31-07-2014

Bugetid/研究經費

1200千元

ResearchField/研究領域

漁業

"珊瑚不管在海洋生物之多樣性，海洋生產力及生態性均是我國至為重要的海洋生物，珊瑚是由珊瑚蟲所構成，具有特殊的生殖策略，珊瑚每年在農曆3 月中旬左右會有集體同時生殖產卵之特性與奇觀。牠們的生殖細胞是如何做到如此精確的生殖週期呢？除了水溫潮汐與光照週期等外在可能因子之影響外，牠們內部的因子(例如生殖激素)如何啓動生殖細胞的發育？這些均是有待探討之重要問題。根據過去本研究室以長期探討雌珊瑚GnRH，aromatase 與sex steroids 之結果及最近預備實驗結果（圖4、圖5 與圖6），而提出珊瑚生殖主軸（圖1）及珊瑚研究假設（圖2 與圖3），本計畫以腎形真葉珊瑚為材料提出研究假設與目的（圖2），本研究目的將針對雌性腎型真葉珊瑚的卵細胞生成作用進行一系列的探討，為了探討珊瑚生殖細胞的發育，我們提出研究策略，選殖重要生殖細胞的標識基因（vasa 與egg protein），並製備抗體，且由研究模式提出外界環境因子（溫度與光照）與內在因子（GnRH 激素與性類固醇激素）對珊瑚生殖細胞發育之影響。本研究計畫將利用3 年期間，利用組織、內分泌、細胞生物及分子遺傳的策略，進行一系列10 項之實驗，包括(1) 探討雌性珊瑚早期生殖細胞的發育特性（early gametogenesis，主要為oogenesis），(2) 選殖珊瑚生殖細胞特有基因（germ cell markers），並進行組織分佈分析，以便進一步探討配子生殖作用，(3) 製備與純化生殖細胞重要蛋白（Vasa 與 Egg protein）的抗體，(4) 探討腎型真葉珊瑚不同生殖季節對雌性生殖細胞標識基因表現之影響，(5) 以原位雜交探討vasa 與egg protein RNA 的存在位置（cellular localization）及在不同生殖階段的表現，(6) 進行抗體免疫染色探討生殖細胞分子標識蛋白存在的位置及在不同生殖階段的表現，(7) 探討GnRH 對雌性珊瑚（in vivo 活體實驗）生殖細胞分子標識表現與生殖細胞發育之影響，(8) 探討GnRH 對雌性珊瑚生殖腺（in vitro 離體實驗）生殖細胞分子標識表現與生殖細胞發育之影響，(9) 探討不同光照周期對雌珊瑚生殖細胞發育與生殖細胞分子標示表現之影響，(10) 探討不同水溫對雌珊瑚生殖細胞發育與生殖細胞分子標示基因表現之影響。經由本研究預期將對珊瑚生殖生物及生殖產卵有深入的瞭解，尤其對生殖細胞如何被啟動發育，發育的機制等關鍵將有所解明，瞭解演化的關係，解答珊瑚集體同時產卵之奧妙。本研究極具學術創新與高度重要性，是屬於國際學界最前緣的研究領域，是國際領先的研究項目，並對珊瑚生殖生理應用及海洋環境生態是十分重要的。" "Corals are very important in marine ecological system in terms of biodiversity, reproduction, and ecology. Corals have special strategies for the reproduction and propagation for their generation. Most broadcast spawning scleractinian corals are synchronously released gametes during a brief annual spawning period. The spawning period is followed the lunar phase (about lunar mid-March in Taiwan). The environmental factors were included temperatures, lunar periodicity, illumination, tidal surge, physical and chemical shock, presence of food, and others. But there were rare studies to elucidate how the endogenous factors may regulate the synchronized gametogenous and mass spawning in female corals. Scleractinian coral Euphyllia ancora is a gonochorism, and spawns in the late spring in Taiwan for external fertilization. According to our previous and preliminary data in female coral, we therefore proposed that environmental cues (photoperiods and temperatures) and endogenous factors (GnRH and sex steroids) to play important roles on oogenesis in corals. We selected five germ cell markers (Vasa and Egg protein) for the study of coral oogenesis. A working model for the coral oogenesis is further proposed in this proposal by selecting female coral, E. ancora as a research species. We intend to answer the following questions in corals. 1) Do germ cell markers (Vasa and Egg protein) also exist in corals as compared to vetebrates? 2) The localization and distribution of germ cell markers (Vasa and Egg protein) in female corals according to in situ hybridization and immunohistochemical staining. 3) The relationship between the expression of germ cell markers and germ cell development. 4) The seasonal effects on the expression of germ cell markers. 5) The photoperiod effects on the expression of germ cell markers and germ cell development. 6) The temperature effects on the expression of germ cell markers and germ cell development. 7) The effects of GnRH on the expression of germ cell markers and germ cell development. 8) The effects of sex steroids on the expression of germ cell markers and germ development. 9) The molecular mechanism of oogenesis in corals."

Keyword(s)

珊瑚
生殖
環境因子
激素
卵細胞生成作用
coral
reproduction cycle
spawning
hormones
oogenesis
environmental cues

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The development of female germ cells in the coral, Euphyllia ancora

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