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| Sequence Analysis and Expression Patterns of miR-10 and Target Gene arhgef3 in Sea Cucumber Apostichopus japonicus Exposed to Salinity Stress |
| GOU Yuqing, WEI Xin, MA Nanxing, LIU Dan, GAO Siqi, TIAN Yi |
| Key Laboratory of Mariculture & Stock Enhancement in North China′s Sea, Ministry of Agriculture and Rural Affairs, College of Fisheries and Life Science, Dalian Ocean University, Dalian 116023, China |
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Abstract In order to investigate the response process of sea cucumber Apostichopus japonicus exposed to salinity stress, differentially expressed miR-10 and the corresponding target gene arhgef3 were screened from the transcriptome and high-throughput sequencing results of miRNAs to analyze the expression of miR-10 and target gene arhgef3 in different tissues of sea cucumber with body weight of (18.55±3.40) g exposed to a salinity of 18 for 0 h, 6 h, 24 h and 48 h with control group at 30 normal salinity to provide a basis for their roles in the salinity response mechanism of the sea cucumber. Analysis of the precursor and mature sequences of miR-10 showed that miR-10 formed a stable stem-loop structure with highly conserved regions in the interval between 29 and 51. The target gene arhgef3 had an open reading frame of 1728 bp encoding 575 amino acids and a functional domain between amino acids 280—500. There was significantly higher expression level of arhgef3 in intestine, respiratory tree and coelomocytes than that in other tissues (P<0.05). Under low salt conditions, the expression level of miR-10 was significantly lower at 24 h and significantly higher at 48 h than at 0 h in the control group, and the expression level of arhgef3 was significantly higher at 6 h of stress than at 0 h in the control group. Transfection of miR-10 inhibitors into coelomocytes was found to be significantly decreased at 24 h and increased at 48 h compared with 0 h in the experimental group, with significantly higher in the experimental group at 0 h at both 6 h and 48 h after transfection with miR-10 mimics than that at 0 h. The expression level of arhgef3 in the experimental group transfected with miR-10 mimic was found to be increased significantly at 24 hours, without significant change in expression level of arhgef3 in the experimental group transfected with miR-10 inhibitor in sea cucumber. The findings indicated that arhgef3 was induced by salinity, and that the target gene was required to maintain a high expression level to adapt to salinity changes and adapt to the low salinity response of sea cucumber during the response to salinity stress.
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Received: 20 October 2022
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