The wide distribution of crustaceans from freshwater through brackish into marine waters and from salt marshes to terrestrial zones shows the exceptional capacities of this animal group to cope with salinity changes Osmoregulation is the main physiological mechanism that maintains the hydromineral homeostasis of these animals The difference of osmoregulation capacities between an osmoconformer and an osmoregulator species is mainly explained by anisosmotic extracellular regulation (AER) and/or isosmotic intracellular regulation (IIR) This review will also discuss the particular acquisition of specific osmoregulation capacities in crustaceans from the first embryonic stages to the adult Several findings revealed that gills and antennal glands are the primordial hydromineral regulation tissues, and they present the same structure and function in all crustaceans, except in shrimps These tissues contain ionocytes that harbor membrane pumps implicated in ion regulation Na+/K+-ATPase, the major of these membrane proteins, acts upon rapid ionic changes, usually in cooperation with several other membrane pumps Also some proteins, belonging to the detoxification and antioxidant systems, seem implicated in the regulation mechanisms after salinity change This review will resume and discuss the current knowledge on salinity regulation in this large and important animal group

Homeostatic responses of crustaceans to salinity changes

Encysted embryos of Artemia franciscana cease development and enter diapause, a state of metabolic suppression and enhanced stress tolerance. The development of diapause-destined Artemia embryos is characterized by the coordinated synthesis of the small heat shock proteins (sHsps) p26, ArHsp21 and ArHsp22, with the latter being stress inducible in adults. The amounts of sHsp mRNA and protein varied in Artemia cysts, suggesting transcriptional and translational regulation. By contrast to p26, knockdown of ArHsp21 by RNA interference had no effect on embryo development. ArHsp21 provided limited protection against stressors such as desiccation and freezing but not heat. ArHsp21 may have a non-essential or unidentified role in cysts. Injection of Artemia adults with amounts of ArHsp22 double-stranded RNA less than those used for other sHsps killed females and males, curtailing the analysis of ArHsp22 function in developing embryos and cysts. The results indicate that diapause-destined Artemia embryos synthesize varying amounts of sHsps as a result of differential gene expression and mRNA translation and also suggest that these sHsps have distinctive functions.

Functional differentiation of small heat shock proteins in diapause-destined Artemia embryos.

Identification, expression pattern, cellular location and potential role of the caveolin-1 gene from Artemia sinica.

Glycogen serves as a metabolic reserve and is involved in macromolecular synthesis. Glycogen phosphorylase (GPase) is a key enzyme involved in intracellular glycogen catabolism, catalyzing the first step in glycogen degradation. In the diapause, GPase catalyzes glycogen into the closely related molecule, sorbitol. In this study, the full-length cDNA of the GPase gene (2,790 bp) was isolated from Artemia sinica for the first time by rapid amplification of cDNA ends technology. The GPase gene encoded a protein of 853 amino acids belonging to the Glycosyltransferase GTB type superfamily. The expression pattern and location of GPase were investigated at various stages during the embryonic development of A. sinica using real-time PCR and in situ hybridization. High GPase expression was detected at the 0 and 5 h stages. Subsequently, expression declined and was maintained at a low level during the stages from 10 to 40 h following by a small increase at day 3. Expression was downregulated at temperatures ranging from 25 to 20 °C and was subsequently upregulated in the range 15–5 °C. In situ hybridization assays showed wide distribution of the GPase gene during different developmental stages. From the results of this study, we conclude that the GPase gene expression is stress-related and might play an important role in Artemia development and metabolism.

Cloning and expression patterns of the brine shrimp (Artemia sinica) glycogen phosphorylase (GPase) gene during development and in response to temperature stress.

Molecular cloning, characterization and expression analysis of the protein arginine N-methyltransferase 1 gene (As-PRMT1) from Artemia sinica

Nuclear protein 1 (NURP1) is a stress-related protein and closely related to diapause in the development of Artemia. In the present paper, the full-length 568-bp cDNA sequence of the nurp1 homolog of Artemia sinica (As-nurp1) was isolated by RACE technology for the first time. The putative As-nurp1 protein consists of 66 amino acids with a basic helix-loop-helix (bHLH) motif and a bipartite nuclear localization signal (NLS). Multiple sequence alignments revealed that the putative As-nurp1 protein sequence was relatively conserved across species, especially in the bHLH domain. The expression of As-nurp1 is widely distributed during A. sinica development. This is followed by a dramatic downregulation after diapause and is newly upregulated from the larval nauplius stage. Furthermore, As-nurp1 transcripts are highly upregulated under conditions of high salinity and low temperature. These findings suggest that As-nurp1 is stress-related and may act as a protective factor in embryonic development.

Molecular Characterization and Expression of As-nurp1 Gene From Artemia sinica During Development and in Response to Salinity and Temperature Stress

As-ClC (chloride channels protein from Artemia sinica), a member from the chloride channels protein family, is a α-helical membrane protein predicted to traverse the cell membrane 11 times. It is important for several physiological functions such as cell volume regulation, cell proliferation, growth and differentiation. In this paper, the complete cDNA sequence of As-CIC was cloned from A. sinica for the first time using RACE technology. The expression pattern and location of the As-CIC gene was investigated in different stages of the embryonic development by means of quantitative real-time PCR and in situ hybridization (ISH) assay. As-CLC was distributed throughout the whole body in cells of different embryonic development of A. sinica as shown by ISH. There was a low expression level of the As-ClC gene after 0 h and a higher expression level after 15 and 40 h when the embryo entered the next growth period and the environmental salinity changed. At adult stage, the As-ClC maintained a high expression level. The results of the real-time PCR assay showed an increasing trend of As-ClC transcripts with increasing salinity. The expression of As-ClC was higher in the control group (28) than in the experimental group except at a salinity of 200 PSU. It indicated that As-ClC functions as salinity-stress-related gene, probably participated in cell volume regulation and osmotic regulation during the early embryonic development of A. sinica.

Expression patterns of As-ClC gene of Artemia sinica in early development and under salinity stress

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Molecular Characterization and Expression of As-nurp1 Gene From Artemia sinica During Development and in Response to Salinity and Temperature Stress

Expression patterns of As-ClC gene of Artemia sinica in early development and under salinity stress