张明霞,
张科,
袁凤娟,
邱渊皓,
冯文坡,
邵向阳,
苏聪颖,
李媛,
董艳美,
王梦琪,
齐金旭,
夏西超^,
平顶山学院医学院, 平顶山 476000

作者简介: 张明霞(1982-),女,硕士,研究方向为生态毒理学,E-mail:626189846@qq.com.

通讯作者: 夏西超,xiaxichao8336@163.com

基金项目: 河南省高等学校重点科研项目（20A330002，19B330002）；河南省联合基金项目（182300410123）；平顶山学院博士启动基金（PXY-BSQD-2018009，PXY-PYJJ-2018005）


中图分类号: X171.5

Characterization of AwCaM1 from Freshwater Clam Anodonta woodiana and Effect of Ca²⁺ and Cd²⁺ on Its Expressions

Zhang Mingxia,
Zhang Ke,
Yuan Fengjuan,
Qiu Yuanhao,
Feng Wenpo,
Shao Xiangyang,
Su Congying,
Li Yuan,
Dong Yanmei,
Wang Mengqi,
Qi Jinxu,
Xia Xichao^,
College of Medicine, Pingdingshan University, Pingdingshan 476000, China

Corresponding author: Xia Xichao,xiaxichao8336@163.com

CLC number: X171.5

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摘要:钙调蛋白（CaM）是Ca²⁺结合蛋白，参与程序性细胞死亡、自噬、肌肉收缩、炎症和免疫应答等多种生物学过程。为探讨Ca²⁺和Cd²⁺对背角无齿蚌（Anodonta woodiana）的生理效应，克隆出AwCaM1全基因序列，分析Ca²⁺和Cd²⁺对AwCaM1表达的影响。结果显示，背角无齿蚌AwCAM1 cDNA全长由692个碱基组成，包含一个516 bp开放阅读框编码的172个氨基酸的多肽链，AwCaM1氨基酸序列包含有4个Ca²⁺结合EF-手形结构域。Ca²⁺处理后，肝胰脏中AwCaM1表达水平在0.01、0.02、0.04和0.08 mg·L^-1处理组呈时间和剂量依赖性方式上调；Cd²⁺处理后，肝胰脏中AwCaM1表达水平在8 mg·L^-1和16 mg·L^-1处理组增加了65.04%（P<0.01）以上。与对照组相比，Ca²⁺处理后鳃中AwCaM1表达水平增加了79.41%（P<0.01）以上，Cd²⁺处理后AwCaM1表达水平增加了88.23%（P<0.01）以上。与对照组相比，外套膜中AwCaM1表达水平在0.16 mg·L^-1的Ca²⁺处理组增加了1.69倍以上（P<0.01），AwCaM1表达水平在8 mg·L^-1和16 mg·L^-1的Cd²⁺处理组增加了1.65倍（P<0.01）以上。以上结果表明，Ca²⁺和Cd²⁺处理对背角无齿蚌AwCaM1表达水平具有显著的诱导作用，其原因与钙的吸收和胁迫效应有关。
关键词: Cd²⁺/
背角无齿蚌/
细胞毒性/
AwCaM1/
Ca²⁺

Abstract:Calmodulin (CaM), a Ca²⁺-binding protein, is involved in programmed cell death, autophagy, muscle contraction, inflammation and the immune response. In order to explore effect of Ca²⁺ and Cd²⁺ on the CaM, one complete cDNA sequence (AwCaM1) was cloned and characterized from the freshwater mussel Anodonta woodiana and its expressions were analyzed. The complete cDNA sequence of AwCaM1 was comprised of 692 bp contained a 516 bp open reading frame which was encoded 172 amino acids. AwCaM1 contained four putative Ca²⁺-binding EF-hand motifs. The AwCaM1 was cloned from the A. woodiana using the rapid amplification of cDNA ends methods and its expression was determined by real-time PCR. In the hepatopancreas, AwCaM1 expression was up-regulated with a time and dose dependent pattern in the Ca²⁺ treated groups (0.01, 0.02, 0.04 and 0.08 mg·L^-1). AwCaM1 mRNA level increased more 65.04% (P<0.01) in the Cd²⁺ treated groups (8 mg·L^-1 and 16 mg·L^-1). In the gill, AwCaM1 expression increased more than 79.41% (P<0.01) compared with that of control group in all the Ca²⁺ treated groups, and more than 88.23% (P<0.01) in all the Cd²⁺ treated groups. In the mantle, AwCaM1 mRNA levels increased more than 1.69 times (P<0.01) in the Ca²⁺ treated group (0.16 mg·L^-1) in contrasted with that of control group, 1.65 times (P<0.01) in Cd²⁺ treated groups (8 mg·L^-1 and 16 mg·L^-1). These results indicated that up-regulations of AwCaM1 expression in bivalve A. woodiana are associated with Ca²⁺ absorption and environmental adaption derived from Ca²⁺ and Cd²⁺ treatment.
Key words:Cd²⁺/
Anodonta woodiana/
cytotoxicity/
AwCaM1/
Ca²⁺.

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