Chinese Journal of Oceanology and LimnologyVol. 33 No. 3, P. 714-724, 2015http://dx.doi.org/10.1007/s00343-015-4143-5 Transcript profi les of mitochondrial and cytoplasmic manganese superoxide dismutases in Exopalaemon carinicauda under ammonia stress* REN Hai (任海) 1, 2 , LI Jian (李健) 2 , ** , LI Jitao (李吉涛) 2 , LIU Ping (刘萍) 2 , LIANG Zhongxiu (梁忠秀) 2 , WU Jianhua (吴建华) 11 Hebei Normal University of Science and Technology, Qinhuangdao 066004, China 2 Key Laboratory of Sustainable Development of Marine Fisheries , Ministry of Agriculture , Yellow Sea Fisheries Research Institute , Chinese Academy of Fishery Sciences , Qingdao 266071 , China Received May 7, 2014; accepted in principle Jul. 22, 2014; accepted for publication Sep. 6, 2014 © Chinese Society for Oceanology and Limnology, Science Press, and Springer-Verlag Berlin Heidelberg 2015 A bstract Superoxide dismutase (SOD) is one of the most important antioxidant defense enzymes, and is considered as the fi rst line against oxidative stress. In this study, we cloned a mitochondrial manganese (Mn) SOD ( m MnSOD ) cDNA from the ridgetail white prawn Exopalaemon carinicauda by using rapid amplifi cation of cDNA ends (RACE) methods. The full-length cDNA for mMnSOD was 1 014-bp long, containing a 5′-untranslated region (UTR) of 37-bp, a 3′-UTR of 321-bp with a poly (A) tail, and included a 657-bp open reading frame encoding a protein of 218 amino acids with a 16-amino-acid signal peptide. The protein had a calculated molecular weight of 23.87 kDa and a theoretical isoelectric point of 6.75. The m MnSOD sequence included two putative N-glycosylation sites (NHT and NLS), the MnSOD signature sequence 180 DVWEHAYY 187 , and four putative Mn binding sites (H48, H96, D180, and H184). Sequence comparison showed that the mMnSOD deduced amino acid sequence of E . carinicauda shared 97%, 95%, 89%, 84%, 82%, 72%, and 69% identity with that of Macrobrachium rosenbergii , M acrobrachium nipponense , Fenneropeneaus chinensis , Callinectes sapidus , P erisesarma b idens , D anio r erio , and Homo sapiens , resectively. Quantitative real-time RT-PCR analysis showed that mMnSOD transcripts were present in all E . carinicauda tissues examined, with the highest levels in the hepatopancreas. During an ammonia stress treatment, the transcript levels of mMnSOD and cMnSOD were up-regulated at 12 h in hemocytes and at 24 h in the hepatopancreas. As the duration of the ammonia stress treatment extended to 72 h, the transcript levels of mMnSOD and cMnSOD signifi cantly decreased both in hemocytes and hepatopancreas. These fi ndings indicate that the SOD system is induced to respond to acute ammonia stress, and may be involved in environmental stress responses in E . carinicauda . Keyword : Exopalaemon carinicauda ; mitochondrial manganese superoxide dismutase ( mMnSOD ); cytosolic manganese superoxide dismutase ( cMnSOD ); cloning; expression 1 INTRODUCTION The ridgetail white prawn Exopalaemon carinicauda is an important cultured economic shrimp species in China (Xu et al., 2010). However, intensifi ed farming and deteriorating conditions in the aquatic environment have resulted in frequent disease outbreaks, for example, “milky shrimp” disease (Xu et al., 2010) and WSSV (Shen et al., 2013). The health of aquatic animals depends on complex interactions among the environment, the pathogen, and the host (Lightner and Redman, 1998). Environmental factors affect not only the abundance of pathogens, but also the disease-resistance of * Supported by the National High Technology Research and Development Program of China (863 Program) (No. 2012AA10A409), the Modern Agro-Industry Technology Research System (No. CARS-47), the Special Fund for Independent Innovation of Shandong Province (No. 2013CX80202), and the Special Fund for Agro-Scientific Research in the Public Interest (No. 201103034) ** Corresponding author: lijian@ysfri.ac.cn
