Molecular cloning and functional characterization of sarco/endoplasmic reticulum Ca 2+ ‐ ATPase from Chinese mitten crab ( Eriocheir sinensis )
- 28 June 2022
- journal article
- research article
- Published by Hindawi Limited in Aquaculture Research
- Vol. 53 (13), 4676-4688
- https://doi.org/10.1111/are.15960
Abstract
Ca2+ plays an important role in signal transduction and regulates various physiological functions of cells. The active transport of Ca2+ from the cytosol to the lumen of the sarcoplasmic or endoplasmic reticulum occurs via sarco/endoplasmic reticulum Ca2+-ATPases (SERCA). However, the function of this protein during moulting cycles remains unclear in Chinese mitten crab (Eriocheir sinensis). In this study, we cloned a cDNA encoding a putative SERCA protein from the Y-organs (moulting glands) in E. sinensis. The cloned Es-SERCA cDNA (4552 bp) included an open reading frame of 3003 bp encoding a 1000-residue protein with a predicted molecular mass of 109.60 kDa. The predicted Es-SERCA protein was clustered with those from other arthropods in phylogenetic analysis. Further, qPCR revealed that Es-SERCA was widely expressed in in all tested tissues with the highest expression in the muscle, followed by the eyestalks, intestines, hepatopancreas, Y-organs and gills. The stage-specific fluctuations among different tissues indicated a functional role of SERCA in calcium homeostasis. The relative mRNA level of the Es-SERCA in Y-organs during the moulting cycle was similar to the level trend of ecdysteroid concentration in haemolymph, indicating that Ca2+ signalling stimulated ecdysteroid synthesis in Y-organs. Altogether, our study provides a reference for further research on the molecular mechanisms of the moulting in E. sinensis.Keywords
This publication has 44 references indexed in Scilit:
- Neuropeptide signaling mechanisms in crustacean and insect molting glandsInvertebrate Reproduction & Development, 2012
- Ecdysteroid metabolism in crustaceansThe Journal of Steroid Biochemistry and Molecular Biology, 2011
- Cloning of sarco-endoplasmic reticulum Ca2+-ATPase (SERCA) from Caribbean spiny lobster Panulirus argusJournal of Comparative Physiology B, 2008
- Morphological and biochemical changes in the muscle of the marine shrimp Litopenaeus vannamei during the molt cycleAquaculture, 2006
- Analysis of Relative Gene Expression Data Using Real-Time Quantitative PCR and the 2−ΔΔCT MethodMethods, 2001
- Cannibalism, refugia and the molting blue crabMarine Ecology Progress Series, 1997
- Deduced amino acid sequence and E1–E2 equilibrium of the sarcoplasmic reticulum Ca2+‐ATPase of frog skeletal muscle Comparison with the Ca2+‐ATPase of rabbit fast twitch muscleFEBS Letters, 1992
- Complementary DNA cloning of a protein highly homologous to mammalian sarcoplasmic reticulum Ca-ATPase from the crustacean ArtemiaJournal of Molecular Biology, 1989
- Two Ca2+ ATPase genes: Homologies and mechanistic implications of deduced amino acid sequencesCell, 1986
- Molt cycle-associated changes in calcium-dependent proteinase activity that degrades actin and myosin in crustacean muscleDevelopmental Biology, 1982