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| Identification of Sodium-calcium Exchanger 1 Gene and Analysis of Its Expression Characteristics Under Cd and Cu Stress in Tegillarca granosa |
| SONG Xing-Rong1,2, ZHANG Yuan1, CHEN Cai-Fang2,*, SHEN Wei-Liang3, LIN Zhi-Hua2 |
1 Zhejiang Key Laboratory of Aquatic Germplasm Resources, College of Biological and Environmental Sciences, Zhejiang Wanli University, Ningbo 315100, China; 2 College of Advanced Agricultural Sciences, Zhejiang Wanli University, Ningbo 315101, China; 3 Ningbo Academy of Oceanology and Fishery, Ningbo 315012, China |
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Abstract As a typical economic bivalve, inhabiting intertidal mudflats, the blood clam (Tegillarca granosa) is prone to selectively accumulate the heavy metal cadmium (Cd). Sodium calcium exchangers (NCXs) are crucial ion transporters on the cell membrane. To explore the role of T. granosa sodium calcium exchanger 1 (TgNCX1) during the selective Cd enrichment process, the blood clams were exposed to 0.089 μmol/L Cd & Cu stress for 120 h. The enrichment characteristic of Cd in T. granosa was detected by inductively coupled plasma-mass spectrometry (ICP-MS), then the sequence features of TgNCX1 were analyzed by the bioinformatics approaches, and its expression profiles were performed by qPCR and Western blot techniques. The results showed that the full-length cDNA of TgNCX1 (GenBank No. PQ255942.1) was 3 001 bp, containing an open reading frame of 2 409 bp and encoding 802 amino acids. Sequence analysis revealed that TgNCX1 belonged to the Calcium/Cation antiporter (CaCA) superfamily, and was a hydrophilic membrane protein with 9 transmembrane domains and 3 Ca2+-binding sites, indicating its capability of binding and transporting Ca2+. Tissue-specific expression analysis showed that TgNCX1 had the highest expression in the gills of T. granosa, suggesting that gill may serve as a critical channel for Cd2+ entry into the organism. The results of temporal expression showed that the expression of TgNCX1 gene and its encoded protein in the gills first increased, and subsequently decreased under Cd & Cu stress. During the early stage of the metal stress, the expression of TgNCX1 gene and its encoded protein increased significantly and peaked at 24 h, which was consistent with Cd enrichment pattern. It was presumed that the reverse transport mode of TgNCX1 may be triggered by Cd2+ competitively binding to the Ca2+-binding domain, driving Cd2+ influx through the electrochemical gradient of Na+, thereby mediating the short-term selective Cd enrichment in T. granosa. After 24 h of Cd & Cu stress, the expression level of TgNCX1 and its encoded protein decreased significantly, while Cd content increased markedly, suggesting that TgNCX1 may not be involved in the long-term selective Cd enrichment of T. granosa, and this process may rely on other transporters or ion channel proteins. This study further reveals the intrinsic regulatory mechanism of selective Cd enrichment, and provide a theoretical basis for cultivating new varieties with low Cd accumulation in T. granosa.
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Received: 14 July 2025
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Corresponding Authors:
*chencaifang@zwu.edu.cn
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