Molecular Biology Reports

, Volume 40, Issue 5, pp 3655–3664 | Cite as

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


As-ClC (chloride channels protein from Artemia sinica), a member from the chloride channels protein family, is a α-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 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.


Expression pattern AS-ClC embryonic development Salt stress Artemia sinica 



We thank anonymous reviewers for valuable comments on an earlier version of the manuscript. This work was supported by the National Science Foundation of China (31071876 and 31272644).


  1. 1.
    Zhu XJ, Feng CZ, Dai ZM, Zhang RC, Yang WJ (2007) AMPK alpha subunit gene characterization in Artemia and expression during development and in response to stress. Stress 10:53–63PubMedCrossRefGoogle Scholar
  2. 2.
    Dai JQ, Zhu XJ, Liu FQ, Xiang JH, Nagasawa H, Yang WJ (2008) Involvement of p90 ribosomal S6 kinase in termination of cell cycle arrest during development of Artemia-encysted embryos. J Biol Chem 283:1705–1712PubMedCrossRefGoogle Scholar
  3. 3.
    Wang JQ, Hou L, Yi N, Zhang RF, Zou XY, Xiao Q, Guo R (2011) Molecular cloning and its expression of trachealess gene (As-trh) during development in brine shrimp, Artemia sinica. Mol Biol Rep 39(2):1659–1665. doi: 10.1007/s11033-011-0905-0 PubMedCrossRefGoogle Scholar
  4. 4.
    Wang JQ, Hou L, Yi N, Zhang RF, Zou XY (2012) Molecular analysis and its expression of a pou homeobox protein gene during development and in response to salinity stress from brine shrimp, Artemia sinica. Comp Biochem Physiol A Physiol 161(1):36–43CrossRefGoogle Scholar
  5. 5.
    Cai Y (1989) A redescription of the brine shrimp (Artemia sinica). Wasmann J Biol 47:105–110Google Scholar
  6. 6.
    Bretag AH (1987) Muscle chloride channels. Physiol Rev 67:618–724PubMedGoogle Scholar
  7. 7.
    Gogelein H (1988) Chloride channels in epithelia. Biochim Biophys Acta 947:521–547PubMedCrossRefGoogle Scholar
  8. 8.
    Liedtke CM (1989) Regulation of chloride transport in epithelia. Annu Rev Physiol 51:143–160PubMedCrossRefGoogle Scholar
  9. 9.
    Worrell RT, Butt AG, Cliff WH, Frizzell RA (1989) A volume-sensitive chloride conductance in human colonic cell line T84. Am J Physiol 256:C1111–C1119PubMedGoogle Scholar
  10. 10.
    Strange K, Denton J (2005) Ste20-type kinases: evolutionarily conserved regulators of ion transport and cell volume. Physiology 21:61–68CrossRefGoogle Scholar
  11. 11.
    Miller C, White MM (1984) Dimeric structure of single chloride channels from Torpedo electroplax. Proc Natl Acad Sci USA 81:2772–2775PubMedCrossRefGoogle Scholar
  12. 12.
    Faraldo-Gómez JD, Roux B (2004) Electrostatics of ion stabilization in a ClC chloride channel homologue from Escherichia coli (clc-5). J Mol Biol 339:981–1000PubMedCrossRefGoogle Scholar
  13. 13.
    Estévez R, Jentsch TJ (2002) CLC chloride channels: correlating structure with function. Curr Opin Struct Biol 12:531–539PubMedCrossRefGoogle Scholar
  14. 14.
    Whiting PJ, Bonnert TP, McKernan RM, Farrar S, Bourdelles LB, Heavens RP, Smith DW, Hewson L, Rigby MR, Sirinathsinghji DJ, Thompson SA, Wafford KA (1999) Molecular and functional diversity of the expanding GABA-A receptor gene family. Ann NY Acad Sci 868:645–653PubMedCrossRefGoogle Scholar
  15. 15.
    Enz R (2001) GABA(C) receptors: a molecular view. Biol Chem 382:1111–1122PubMedCrossRefGoogle Scholar
  16. 16.
    Jentsch TJ, Stein V, Weinreich F, Zdebik AA (2002) Molecular structure and physiological function of chloride channels. Physiol Rev 82:503–568PubMedGoogle Scholar
  17. 17.
    Bradbury NA (1999) Role of intracellular CFTR in acidification. Physiol Rev 79:S175–S191PubMedGoogle Scholar
  18. 18.
    Sheppard DN, Welsh MJ (1999) Structure and function of the CFTR chloride channel. Physiol Rev 79:S23–S45PubMedGoogle Scholar
  19. 19.
    Purdy MD, Wiener MC (2000) Expression, purification, and initial structural characterization of YadQ, a bacterial homolog of mammalian ClC chloride channel proteins. FEBS Lett 466:26–28PubMedCrossRefGoogle Scholar
  20. 20.
    Estévez R, Pusch M, Ferrer-Costa C, Orozco M, Jentsch TJ (2004) Functional and structural conservation of CBS domains from CLC chloride channels. J Physiol 557:363–378PubMedCrossRefGoogle Scholar
  21. 21.
    Meyer S, Savaresi S, Forster IC, Dutzler R (2007) Nucleotide recognition by the cytoplasmic domain of the human chloride transporter ClC-5. Nat Struct Mol Biol 14:60–67PubMedCrossRefGoogle Scholar
  22. 22.
    Okada Y, Maeno E (2001) Apoptosis cell volume regulation and volume-regulatory chloride channels. Comp Biochem Physiol A Physiol 130:377–383CrossRefGoogle Scholar
  23. 23.
    Suzukia M, Moritaa T, Iwamotob T (2006) Diversity of Cl channels. Cell Mol Life Sci 63:12–24CrossRefGoogle Scholar
  24. 24.
    Hou L, Jiang LJ, Sun WJ, Zhang RF, Wang JQ, Zhao XT, An JL (2006) Establishment and improvement of real-time fluorescence quantitative PCR for actin gene of Artemia sinica. J Liaoning Normal Univ 29:15–19Google Scholar
  25. 25.
    Sun PS, Soderlund M, Venzon NC, Ye D, Lu Y (2007) Isolation and characterization of two actins of the Pacific white shrimp, Litopenaeus vannamei. Mar Biol 151:2145–2151CrossRefGoogle Scholar
  26. 26.
    Mindell JA, Maduke M (2001) ClC chloride channels. Genome Biol 2:3003.1–3003.6CrossRefGoogle Scholar
  27. 27.
    Sontheimer H (2003) Malignant gliomas: perverting glutamate and ion homeostasis for selective advantage. Trends Neurosci 26:543PubMedCrossRefGoogle Scholar
  28. 28.
    Saviane C, Conti F, Pusch M (1999) The muscle chloride channel ClC-1 has a double-barreled appearance that is differentially affected in dominant and recessive myotonia. J Gen Physiol 113:457–468PubMedCrossRefGoogle Scholar
  29. 29.
    Shan X, Dunbrack RL, Christopher SA, Kruger WD (2001) Mutations in the regulatory domain of cystathionine beta synthase can functionally suppress patient-derived mutations in cis. Hum Mol Genet 10:635–643PubMedCrossRefGoogle Scholar
  30. 30.
    Bowne SJ, Sullivan LS, Blanton SH, Cepko CL, Blackshaw S, Birch DG, Hughbanks-Wheaton D, Heckenlively JR, Daiger SP (2002) Mutations in the inosine monophosphate dehydrogenase 1 gene (IMPDH1) cause the RP10 form of autosomal dominant retinitis pigmentosa. Hum Mol Genet 11:559–568PubMedCrossRefGoogle Scholar
  31. 31.
    Bianchi L, Miller DM, George AL (2001) Expression of a ClC chloride channel in Caenorhabditis elegans gamma-aminobutyric acid-ergic neurons. Neurosci Lett 299:177–180PubMedCrossRefGoogle Scholar
  32. 32.
    Bikfalvi A, Savona C, Perollet C, Javerzat S (1998) New insights in the biology of fibroblast growth factor-2. Angiogenesis 1:155–173PubMedCrossRefGoogle Scholar
  33. 33.
    Adams TE, McKern NM, Ward CW (2004) Signalling by the type 1 insulin-like growth factor receptor: interplay with the epidermal growth factor receptor. Growth Factors 22:89–95PubMedCrossRefGoogle Scholar
  34. 34.
    Tallquist M, Kazlauskas A (2004) PDGF signaling in cells and mice. Cytokine Growth Factor Rev 15:205–213PubMedCrossRefGoogle Scholar
  35. 35.
    Valenzuela SM, Mazzanti M, Tonini R, Qiu MR, Warton K, Musgrove EA, Campbell TJ, Breit SN (2000) The nuclear chloride ion channel NCC27 is involved in regulation of the cell cycle. J Physiol 529:541–552PubMedCrossRefGoogle Scholar
  36. 36.
    Shen MR, Droogmans G, Eggermont J, Voets T, Ellory JC, Nilius B (2000) Differential expression of volume—regulated anion channels during cell cycle progression of human cervical cancer cells. J Physiol 529:385–394PubMedCrossRefGoogle Scholar
  37. 37.
    Nilius B (2001) Chloride channels go cell cycling. J Physiol 532:581PubMedCrossRefGoogle Scholar
  38. 38.
    Wondergem R, Gong W, Monen SH, Dooley SN, Gonce JL, Conner TD, Houser M, Ecay TW, Ferslew KE (2001) Blocking swelling-activated chloride current inhibits mouse liver cell proliferation. J Physiol 532:661–672PubMedCrossRefGoogle Scholar
  39. 39.
    Li M, Wang B, Lin W (2008) C-l channel blockers inhibit cell proliferation and arrest the cell cycle of hum an ovarian cancer cells. Eur J Gynaecol Oncol 29(3):267–271PubMedGoogle Scholar
  40. 40.
    Kunzelmann K (2005) Ion channels and cancer. J Membr Biol 205:159–173PubMedCrossRefGoogle Scholar
  41. 41.
    Bachmann O, Heinzmann MA, Manns MP, Seidler U (2007) Mechanisms of secretion associated shrinkage and volume recovery in cultured rabbit parietal cells. Am J Physiol Gastrointest Liver Physiol 292:G711–G717PubMedCrossRefGoogle Scholar
  42. 42.
    Zhang HN, Zhou JG, Qiu QY, Ren JL, Guan YY (2006) ClC-3 chloride channel prevents apoptosis induced by thapsigargin in PC12 cells. Apoptosis 11:327–336PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media Dordrecht 2012

Authors and Affiliations

  • Qiaozhi Zhang
    • 1
  • Ming Hou
    • 2
  • Qiuying Li
    • 1
  • Lulu Han
    • 1
  • Zhe Yuan
    • 1
  • Jian Tan
    • 1
  • Bin Du
    • 1
  • Xiangyang Zou
    • 3
  • Lin Hou
    • 1
    • 4
  1. 1.College of Life SciencesLiaoning Normal UniversityDalianPeople’s Republic of China
  2. 2.The Affiliated HospitalChangchun University of Traditional Chinese MedicineChangchunPeople’s Republic of China
  3. 3.Department of BiologyDalian Medical UniversityDalianPeople’s Republic of China
  4. 4.College of Life SciencesLiaoning Normal UniversityDalianPeople’s Republic of China

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