title: Identification and characterization of the cDNAs encoding the two subunits of Chinese mitten crab (Eriocheir sinensis) calcineurin: their implications in stress and immune response.
abstract: Calcineurin (CN), the only Ca(2+)/calmodulin-activated serine/threonine protein phosphatase, is a key effector participating in Ca(2+)-dependent signal transduction pathways in a number of cellular processes under normal, stress and pathological conditions. However, the expression and the relevance of CN in stress and immune response have not been characterized in crustaceans. Here, we identified the cDNAs that encode the two subunits of CN (termed EsCN-A and EsCN-B, respectively) in Chinese mitten crab Eriocheir sinensis and analysed their expression patterns in response to stress and immune challenges. The catalytic subunit EsCN-A is comprised of 511 amino acids with a theoretical molecular mass of 57.5 kDa and shows 80% sequence identity with human beings CN-A alpha isoform, while the regulatory subunit EsCN-B protein is composed of 170 amino acids with an estimated molecular mass of 19.3 kDa and shares 88% sequence identity with human beings CN-B type 1. Tissue distribution analysis reveals that both EsCN-A and EsCN-B mRNA transcripts are expressed in all tested tissues with the greatest expression in hepatopancreas and the lowest expression in haemocytes. In addition, both EsCN-A and EsCN-B genes could be significantly up-regulated but with different expression patterns by ambient salinity (15‰ and 30‰ salinities) and pH (pH 6 and 8.5) stresses in gill, hepatopancreas, haemocytes, intestine and muscle. Furthermore, EsCN-A and EsCN-B were up-regulated by LPS and Poly(I:C) immune stimulations in E. sinensis haemocytes in vitro. Moreover, EsCN-A and EsCN-B mRNA were significantly up-regulated in haemocytes, gill, hepatopancreas, intestine and muscle in response to Edwardsiella tarda challenge in vivo. Finally, we revealed the importance of EsCN in LPS-induced nitric oxide production in E. sinensis haemocytes. Together our observations suggest that EsCN, the important downstream effector of CaM-mediated signalling pathway(s), may possess vital roles in stress and immune response in the Chinese mitten crab.
authors.affiliation: Tianjin Key Laboratory of Animal and Plant Resistance, College of Life Sciences, Tianjin Normal University, 393 West Binshui Road, Xiqing District, Tianjin 300387, PR China. Electronic address: shuo76@yahoo.com.
authors.forename: Shuo
authors.initials: S
authors.lastname: Li
authors.affiliation: Tianjin Key Laboratory of Animal and Plant Resistance, College of Life Sciences, Tianjin Normal University, 393 West Binshui Road, Xiqing District, Tianjin 300387, PR China.
authors.forename: Zirui
authors.initials: Z
authors.lastname: Jia
authors.affiliation: Tianjin Key Laboratory of Animal and Plant Resistance, College of Life Sciences, Tianjin Normal University, 393 West Binshui Road, Xiqing District, Tianjin 300387, PR China.
authors.forename: Xiaoli
authors.initials: X
authors.lastname: Chen
authors.affiliation: Tianjin Center for Control and Prevention of Aquatic Animal Infectious Disease, 442 South Jiefang Road, Hexi District, Tianjin 300221, PR China.
authors.forename: Xuyun
authors.initials: X
authors.lastname: Geng
authors.affiliation: Tianjin Key Laboratory of Animal and Plant Resistance, College of Life Sciences, Tianjin Normal University, 393 West Binshui Road, Xiqing District, Tianjin 300387, PR China. Electronic address: jssun1965@aliyun.com.
authors.forename: Jinsheng
authors.initials: J
authors.lastname: Sun
mesh_terms: D000276:Adjuvants, Immunologic / Q000494:pharmacology; D000595:Amino Acid Sequence; D000818:Animals; D060829:Arthropod Proteins / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D001483:Base Sequence; D003386:Brachyura / Q000187:drug effects / Q000235:genetics* / Q000276:immunology* / Q000378:metabolism; D019703:Calcineurin / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D018076:DNA, Complementary / Q000235:genetics / Q000378:metabolism; D020609:Edwardsiella tarda / Q000502:physiology; D005786:Gene Expression Regulation; D007113:Immunity, Innate / Q000187:drug effects; D008070:Lipopolysaccharides / Q000494:pharmacology; D008969:Molecular Sequence Data; D011070:Poly I-C / Q000494:pharmacology; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D054712:Salinity; D016415:Sequence Alignment; D012965:Sodium Chloride / Q000494:pharmacology; D013312:Stress, Physiological / Q000187:drug effects
chemical_list: D000276:Adjuvants, Immunologic; D060829:Arthropod Proteins; D018076:DNA, Complementary; D008070:Lipopolysaccharides; D012333:RNA, Messenger; D012965:Sodium Chloride; D019703:Calcineurin; D011070:Poly I-C
keywords: Calcineurin; Chinese mitten crab; Environmental stresses; Eriocheir sinensis; Immune response