Calmodulin Highly Expressed during the Formation of Pearl Sac in Freshwater Pearl Mussel (Hyriopsis schlegelii)

Kou Peng1,2, Fang-lan Liu1,2, Jun-hua Wang1,2, Yi-jiang Hong1,2
1School of Life Sciences, Nanchang University, Nanchang, China
2Key Laboratory of Aquatic animals Resources and Utilization of Jiangxi, Nanchang University, Jiangxi, China

Tóm tắt

Calmodulin (CaM) is a pivotal multifunction regulator of calcium metabolism in nearly all organisms, and earlier studies revealed its underlying involvement of shell or pearl formation in oysters. Culture pearls are formed within complete pearl sacs. However, whether CaM functions in pearl sac formation or not is not reported. In this study, we prepared a polyclonal antibody against the CaM from the freshwater pearl mussel Hyriopsis schlegelii (HsCaM) and then detected the expression and distribution of HsCaM in pearl sac using the method of immunofluorescence in situ hybridization (IISH). Results revealed that strong fluorescent signals were observed in pearl sac as well as epithelial border of mantle. In addition, quantitative analysis results indicated that the mRNA expression level of HsCaM in pearl sac of H. schlegelii was significantly up-regulated during pearl sac formation. The fact that HsCaM highly expressed is present in pearl sac suggested that it might facilitate pearl sac formation in H. schlegelii.

Tài liệu tham khảo

Arnaud-Haond S, Goyard E, Vonau V, Herbaut C, Prou J, Saulnier D (2007) Pearl formation: persistence of the graft during the entire process of biomineralization. Mar Biol 9(1):113–116 Bai Z, Zheng H, Lin J, Wang G, Li J (2013) Comparative analysis of the transcriptome in tissues secreting purple and white nacre in the pearl mussel Hyriopsis cumingii. PLoS One 8(1):e53617 Cheung WY (1980) Calmodulin plays a pivotal role in cellular regulation. Science 207(4426):19–27 Eddy L, Affandi R, Kusumorini N, Sani Y, Manalu W (2015) The pearl sac formation in male and female Pinctada maxima host oysters implanted with allograft saibo. Hayati J Biotechnol 22(3):122–129 Fang Z, Yan Z, Li S, Wang Q, Cao W, Xu G, Xiong X, Xie L, Zhong R (2008a) Localization of calmodulin and calmodulin-like protein and their functions in biomineralization in P. fucata. Prog Nat Sci 18:405–412 Fang Z, Cao W, Li S, Wang Q, Li C, Xie L, Zhang R (2008b) Significance of the C-terminal globular domain and the extra tail of the calmodulin-like protein (Pinctada fucata) in subcellular localization and protein e protein interaction. Cell Biol Int 32:920–927 Fiske D, Shepherd J (2007) Continuity and change in Chinese freshwater pearl culuture. Gems and Gemology 43(2):138–145 He SH, Peng K, Hong YJ, Wang JH, Sheng JQ, Gu Q (2013) Molecular properties and immune defense of two ferritin subunits from freshwater pearl mussel, Hyriopsis schlegelii. Fish Shellfish Immunol 34(3):865–874 Huang J, Zhang C, Ma Z, Xie L, Zhang R (2007) A novel extracellular EF-hand protein involved in the shell formation of pearl oyster. Biochim Biophys Acta 1770(7):1037–1044 Inoue N, Ishibashi R, Ishikawa T, Atsumi T, Aoki H, Komaru A (2011a) Gene expression patterns in the outer mantle epithelial cells associated with pearl sac formation. Marine. Biotechnology 13(3):474–483 Inoue N, Ishibashi R, Ishikawa T, Atsumi T, Aoki H, Komaru A (2011b) Can the quality of pearls from the Japanese pearl oyster (Pinctada fucata) be explained by the gene expression patterns of the major shell matrix proteins in the pearl sac? Marine. Biotechnology 13(1):48–55 Ji P, Yao C, Wang Z (2011) Two types of calmodulin play different roles in Pacific white shrimp (Litopenaeus vannamei) defenses against Vibrio Parahaemolyticus and WSSV infection. Fish Shellfish Immunol 31:260–268 Kinoshita S, Wang N, Inoue H, Maeyama K, Okamoto K, Nagai K, Kondo H, Hirono I, Asakawa S, Watabe (2011) Deep sequencing of ESTs from nacreous and prismatic layer producing tissues and a screen for novel shell formation-related genes in the pearl oyster. PLoS One 6(6): e21238 Kishore P, Southgate P (2015) Development and function of pearl-sacs grown from regenerated mantle graft tissue in the black-lip pearl oyster, Pinctada margaritifera (Linnaeus, 1758). Fish Shellfish Immunol 45:567–573 Li S, Xie L, Zhang C, Zhang Y, Gu M, Zhang R (2004) Cloning and expression of a pivotal calcium metabolism regulator: calmodulin involved in shell formation from pearl oyster (Pinctada fucata). Comp Biochem Physiol Part B: Biochem Mol Biol 138(3):235–243 Li S, Xie L, Ma Z, Zhang R (2005) cDNA cloning and characterization of a novel calmodulin-like protein from pearl oyster Pinctada fucata. FEBS J 272(19):4899–4910 Liu X, Li J (2015) Formation of the prismatic layer in the freshwater bivalve Hyriopsis cumingii: the feedback of crystal growth on organic matrix. Acta Zool (Stockholm) 96:30–36 Liu X, Li J, Xiang L, Sun J, Zheng G, Zhang G, Wang H, Xie L, Zhang R (2012) The role of matrix proteins in the control of nacreous layer deposition during pearl formation. Proc R Soc B Biol Sci 279:1000–1007 Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using real-time quantitative PCR and the 2(−Delta Delta C(T)) method. Methods 25(4):402–408 McGinty EL, Evans BS, Taylor JU, Jerry DR (2010) Xenografts and pearl production in two pearl oyster species, P. maxima and P. margaritifera: effect on pearl quality and a key to understanding genetic contribution. Aquaculture 302(3–4):175–181 Means AR, Dedman JR (1980) Calmodulin-an intracellular calcium receptor. Nature 285(5760):73–77 Peng K, Wang JH, Sheng JQ, Zeng LG, Hong YJ (2012a) Molecular characterization and immune analysis of a defensin from freshwater pearl mussel, Hyriopsis schlegelii. Aquaculture 334-337:45–50 Peng K, Wang JH, Liu TT, Sheng JQ, Shi JW, Shao P, He SH, Hong YJ (2012b) Expression analysis and immune response of the cathepsin L from freshwater pearl mussel, Hypriopsis schlegelii. Acta Hydrobiologica Sinca 36(6):1128–1134 in Chinese Peng K, Wang CY, Wang JH, Sheng JQ, Shi JW, Li J, Hong YJ (2014) Molecular cloning, sequence analysis, and cadmium stress-rated expression changes of BTG1 in freshwater pearl mussel, Hyriopsis schlegelii. Zool Res 35(5):389–397 Peng K, Hu B, Zhao D, Wang J, Sheng J, Wu D, Shi J, Hong YJ (2016) Molecular characterization and expression analysis of Cathepsin L1-like from Hyriopsis schlegelii. J Fish China 40(10):1556–1566 in Chinese Ren G, Hu X, Tang J, Wang Y (2013) Characterization of cDNAs for calmodulin and calmodulin-like protein in the freshwater mussel Hyriopsis cumingii: differential expression in response to environmental Ca(2+) and calcium binding of recombinant proteins. Comp Biochem Physiol B Biochem Mol Biol 165(3):165–171 Zeng LG, Wang JH, Li YJ, Sheng JQ, Gu Q, Hong YJ (2012) Molecular characteristics and expression of calmodulin cDNA from the freshwater pearl mussel, Hyriopsis schlegelii. Genet Mol Res 11(1):42–52