Expression patterns of As - ClC gene of Artemia sinica in early development and under salinity stress
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Expression patterns of As-ClC gene of Artemia sinica in early development and under salinity stress Qiaozhi Zhang • Ming Hou • Qiuying Li • Lulu Han • Zhe Yuan • Jian Tan • Bin Du Xiangyang Zou • Lin Hou
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Received: 28 August 2012 / Accepted: 18 December 2012 / Published online: 1 January 2013 Ó Springer Science+Business Media Dordrecht 2012
Abstract As-ClC (chloride channels protein from Artemia sinica), a member from the chloride channels protein family, is a a-helical membrane protein predicted to traverse the cell membrane 11 times. It is important for several physiological functions such as cell volume regulation, cell proliferation, growth and differentiation. In this paper, the complete cDNA sequence of As-CIC was cloned from A. sinica for the first time using RACE technology. The expression pattern and location of the As-CIC gene was investigated in different stages of the embryonic development by means of quantitative real-time PCR and in situ hybridization (ISH) assay. As-CLC was distributed throughout the whole body in cells of different embryonic development of A. sinica as shown by ISH. There was a low expression level of the As-ClC gene after 0 h and a higher expression level after 15 and 40 h when the embryo entered the next growth period and the environmental salinity changed. At adult stage, the As-ClC maintained a high Q. Zhang Q. Li L. Han Z. Yuan J. Tan B. Du L. Hou College of Life Sciences, Liaoning Normal University, Dalian 116029, People’s Republic of China M. Hou The Affiliated Hospital, Changchun University of Traditional Chinese Medicine, Changchun 130021, People’s Republic of China X. Zou (&) Department of Biology, Dalian Medical University, Dalian 116044, People’s Republic of China e-mail: [email protected] L. Hou (&) College of Life Sciences, Liaoning Normal University, 1, Liushu South Street, Ganjingzi District, Dalian 116081, People’s Republic of China e-mail: [email protected]
expression level. The results of the real-time PCR assay showed an increasing trend of As-ClC transcripts with increasing salinity. The expression of As-ClC was higher in the control group (28) than in the experimental group except at a salinity of 200 PSU. It indicated that As-ClC functions as salinity-stress-related gene, probably participated in cell volume regulation and osmotic regulation during the early embryonic development of A. sinica. Keywords Expression pattern AS-ClC embryonic development Salt stress Artemia sinica
Introduction The brine shrimp Artemia is a small crustacean living in high salinity water worldwide and possesses powerful adaptability to extreme environments including hypersalinity, anoxia and temperature [1, 2]. Therefore, Artemia has evolved a unique developmental pattern of encysted diapausing embryos to cope with various extreme environments [2]. However, the natural populations of A. sinica in China in salt pools or brine pits have declined dramatically due to the dry out of their ephemeral natural habitats [3]. A decline in brine shrimp populations could
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