Evaluation of Mitochondrial DNA Molecules in Cryoprotectant and Chilling Treated Hard Coral (Echinopora Spp.) Oocytes

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Project title

Code/計畫編號

NSC101-2313-B291-002

Translated Name/計畫中文名

抗凍劑對低溫保存石珊瑚(Echinopora spp.)卵粒腺體DNA分子影響之研究

Project Coordinator/計畫主持人

Jiann-Chu Chen

Funding Organization/主管機關

National Science and Technology Council

Co-Investigator(s)/共同執行人

林家興

Department/Unit

National Museum of Marine Biology and Aquarium Planning & Research Division

Website

https://www.grb.gov.tw/search/planDetail?id=2636010

Year

2012

Start date/計畫起

01-08-2012

Expected Completion/計畫迄

31-07-2013

Bugetid/研究經費

800千元

ResearchField/研究領域

海洋科學
生物技術(理)

目前本實驗室在珊瑚卵之冷凍保存已有初步研究成果 (Tsai et al. 2010, Tsai et al. 2011) ，其中包括探討不同抗凍劑對於低溫保存珊瑚卵的影響，其研究發現珊瑚卵的三磷酸腺苷及粒線體膜電位受到冷凍因子的改變而下降，經共軛焦顯微鏡圖像顯示珊瑚卵內粒腺體分佈模式異常推測抗凍劑致使粒腺體受損。由於三磷酸腺苷之合成主要由電子傳遞鏈中的相關組成成份負責，其組成成份主要是由細胞核及粒腺體轉譯及轉錄而來。因此，進一步了解抗凍劑及低溫保存對於粒線體DNA所造成的傷害尤其重要。在本研究中，我們將利用real-time PCR的技術完成珊瑚卵 (Echinopora spp.) 粒線體DNA分子在不同冷凍保存條件下的分析，並與三磷酸腺苷合成的結果進行比較，最後對珊瑚粒腺體基因組進行新一代定序序列分析 (NGS)，以利日後進行珊瑚基因表現之相關研究。本研究成果將以分子生物學層次的觀點加以探討，進一步了解冷凍保存相關因子對於珊瑚卵之影響，並提供有系統及實質的珊瑚復育成效。 We previously reported that cryoprotectants and low temperature preservation damage coral mitochondrial distribution and membrane potential resulting in reduced adenosine triphosphate (ATP) production (Tsai et al., 2010, Tsai et al., 2011). The components of the electron transport chain (ETC), which is responsible for the majority of a cell’s ATP production, are encoded by both the nuclear and the mitochondrial genomes. Hence, efficient ETC function for producing maximum cellular ATP requires transcription and translation of both nuclear and mitochondrial genomes. As mitochondrial DNA molecule encodes the components of the ETC and plays a crucial role in the capacity for ATP synthesis, we will determine the effect of cryoprotectants and low temperature on mitochondrial DNA molecule in hard coral (Echinopora spp.) oocytes using quantitative real-time PCR technique. Detection of mt-DNA content is valuable in assessing oocyte quality. This is particularly important in spawning coral species where direct measurement of in vitro fertilization and developmental ability has not yet been possible. The findings will provide direct evidence if hard coral (Echinopora spp.) oocytes are extremely susceptible to cryoprotectants or low temperature preservation and support concerns on the adverse effects of cryoprotectants.

Keyword(s)

低溫保存
抗凍劑
粒線體DNA
定量PCR
石珊瑚
mitochondriamitochondriamitochondriamitochondriamitochondriamitochondriamitochondriamitochondriamitochondria mitochondriamitochondrial DNA
hard coral
real-time PCR
cryoprotectant
low temperature preservation

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Evaluation of Mitochondrial DNA Molecules in Cryoprotectant and Chilling Treated Hard Coral (Echinopora Spp.) Oocytes

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