The oriental river prawn Macrobrachium nipponense is an important aquaculture species in China, Vietnam, and Japan. This species could survive in the salinity ranging from 7 to 20 ppt and accelerate growth in the salinity of 7 ppt. To identified the genes and pathways in response to acute high salinity stress, M. nipponense were exposed to the acute high salinity of 25 ppt. Total RNA from hepatopancreas, gills, and muscle tissues was isolated, and then sequenced using high throughput sequencing method. Differentially expressed genes (DGEs) were identified, and total of 632, 836, and 1246 DEGs with a cutoff of significant two-fold change were differentially expressed in hepatopancreas, gills, and muscle tissues, respectively. Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genome pathway enrichment analysis were conducted. These DEGs were involved in the GO terms of cellular process, metabolic process, membrane, organelle, binding, and catalytic activity. The DEGs of hepatopancreas and gill tissues were mainly enriched in PPAR signaling pathway, longevity regulating pathway, protein digestion and absorption, and the DEGs of muscle tissue in arginine biosynthesis, adrenergic signaling in cardiomyocytes, cardiac muscle contraction, and cGMP-PKG signaling pathway. Real-time PCR conducted with fifteen selected DEGs indicated high reliability of digital analysis using RNA-Seq. This work provides a comprehensive insight into the molecular responses to high salinity stress in M. nipponense, which provide a novel contribution to understanding of the molecular mechanisms of adaptation to salinity stress in euryhaline crustaceans.
Salinity is an ecological factor affecting the physiology, survival, and distribution of crustaceans. Additionally, salinity fluctuation detrimentally affects the composition and biological process of crustaceans. As a significant commercial aquaculture species in China, Japan, and Southeast Asian countries, the oriental river prawn, Macrobrachium nipponense, can tolerate a wide range of salinity. The transcriptome, proteome, histology, and physiology analysis were utilized to explore the physiological responses and molecular mechanisms of salinity tolerance in M. nipponense. Through the three-month culture, the statistic of growth trait illustrated the relatively excellent performance of M. nipponense in low salinity, and the higher salinity exposure significantly affected the growth of M. nipponense. In terms of the histological analysis, the gills and hepatopancreas of M. nipponense suffered varying degrees of damage. Besides, the activities of the digestive, immune-related, and metabolic enzymes were calculated. These results indicated that salinity significantly influenced trypsin and amylase in hepatopancreas, especially in 14 ppt. The immune-related enzymes were activated in high salinity. Notably, the activity of metabolic enzymes was significantly low in 7 and 14 ppt, which testified that the 7 ppt to 14 ppt were near the isotonic point of M. nipponense. In gills, hepatopancreas, and muscle, high-throughput mRNA sequencing revealed 11356, 2227, and 1819 differentially expressed genes (DEGs) by comparing the 7, 14, and 21 ppt groups with the 0ppt group, respectively. The TMT-labeling proteome identified 439 and 230 differentially expressed proteins (DEPs) in gills and hepatopancreas through the comparison of the 7, 14, and 21 ppt groups to the 0 ppt group, respectively. Additionally, through the integration of transcriptome and proteome, several pathways related to salinity adaptation were enriched, including protein export, cGMP-PKG signaling pathway, Amino sugar and nucleotide sugar metabolism, and Glycine, serine and threonine metabolism. Besides, 16 up and down-regulated proteins and related DEGs were detected through KEGG enrichment analysis, including ETHE1, BIP, chitinase (E3.2.1.14), and SARDH. Notably, no significantly regulated proteins and related DEGs were recorded by the correlation of transcriptome and proteome of 0 ppt and 7 ppt in hepatopancreas. Thus, the optimum survival salinity of M. nipponense may range from 0 ppt to 7 ppt. Overall, these results may provide valuable insights into the mechanisms underlying the culture of M. nipponense in different salinity.
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