Isolation of tissue layers in hermatypic corals by N-acetylcysteine: morphological and proteomic examinations |
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Authors: | S-E Peng Y-J Luo H-J Huang I-T Lee L-S Hou W-N U Chen L-S Fang C-S Chen |
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Institution: | (1) Institute of Marine Biotechnology, National Dong Hwa University, 2 Houwan Road, Checheng, Pingtung, 944, Taiwan, ROC;(2) National Museum of Marine Biology & Aquarium, 2 Houwan Road, Checheng, Pingtung, 944, Taiwan, ROC;(3) Department of Biotechnology, I-Sou University, Kaohsiung, Taiwan, ROC;(4) Cheng Shiu University, Niao-Song, Taiwan, ROC |
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Abstract: | Corals are diploblastic in body pattern and include two tissue layers, the epidermis and gastrodermis, interconnected by an
acellular matrix mesoglea. During development, cells in these tissue layers differentiate morphologically and functionally.
In most hermatypic corals, the gastrodermis further develops an ability to associate with microalgae dinoflagellates. This
endosymbiosis occurs inside specific host gastrodermal cells, and its mechanism still remains unclear notwithstanding decades
of research. The delay in progress is partly due to the difficulty in separating the gastrodermis and its symbionts from the
epidermis for detailed cellular and biochemical investigations. The present study reports a simple method to separate these
two tissue layers in hermatypic corals using the reducing agent, N-acetylcysteine (NAC). Efficient tissue and proteomic isolations are demonstrated by microscopy and two-dimensional SDS polyacrylamide
gel electrophoresis (2D SDS-PAGE). The NAC treatment was able to separate tissue layers without inducing protein degradation.
Furthermore, the sensitivity of protein detection greatly increases in the isolated tissue layers. The application of the
present technique provides future research on endosymbiosis and coral development with a tool for higher accuracy and sensitivity. |
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Keywords: | Epidermis Gastrodermis Proteomics Endosymbiosis Symbiodinium |
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