The full-length complementary (c)DNA of vacuolar-type-H(+) -ATPase B1 gene (vhab1) in marbled eel Anguilla marmorata with 1741 base pairs (bp) was identified. It contained a 1512 bp open reading frame encoding a polypeptide with 503 amino acids (55·9 kDa), an 83 bp 5'-untranslated region (UTR) and a 146 bp 3'-UTR. The expression levels of A. marmorata vhab1 in gill and kidney of A. marmorata were evaluated at different intervals during the exposure to various salinities (0, 10 and 25). The results indicated that the expression levels of A. marmorata vhab1 messenger (m)RNA in gill and kidney had a significant increase and reached the highest level at 1 h in brackish water (BW, salinity 10) group and 6 h in seawater (SW, salinity 25) group. Therefore, salinity did affect the relative expression level of A. marmorata vhab1 mRNA in gills, which exhibited the enhancement by c. 44 times in SW group when compared with that in fresh water. No remarkable difference in the expression of A. marmorata vhab1 mRNA was observed after 15 days of SW exposure (P > 0·05). V-H(+) -ATPase activity exhibited an increase by two- to three-fold when compared with that in gill and kidney from the control group. The consequence primarily suggested that A. marmorata vhab1 gene product in elvers from A. marmorata plays an important role in adaptation response to SW.
ABSTRACT. Two heat-shock protein (HSP) 70 family transcripts, heat-shock protein 70 cognate 5 and heat-shock protein 70 cognate 3 (designated as EsHSC70-5 and EsHSC70-3, respectively), were isolated from the Chinese mitten crab Eriocheir sinensis and their expression profiles were evaluated for their responsiveness to larval development and immune challenge in adult crabs. The HSPs exhibited 45-89% identity with other heat-shock proteins, and they shared similar structural features. EsHSC70 mRNA expression was detected not only during infection but also during the developmental larval stages. The EsHSC70s were enriched, and their expression fluctuated during early development. EsHSC70 mRNA expression was significantly induced by Vibrio parahaemolyticus challenge in all of the tissues studied (P < 0.05). Expression of EsHSC70 mRNA in the hepatopancreas and at the early zoeal stages was particularly pronounced, and the two EsHSC70s exhibited differential expression patterns both chronologically and spatially. The EsHSC70-5 mRNA level was significantly downregulated in the intestine and gills compared to that in controls at nearly all time points, and was expressed at a lower level after the bacterial challenge, indicating that EsHSC70-5 and EsHSC70-3 respond to immune challenges. The stage-specific enrichment of EsHSC70 transcripts in crabs suggests that these stress proteins play an essential role during brachyurization events.
In arthropods, the moult-inhibiting hormone (MIH), the ecdysone receptor (EcR), and the retinoid X receptor (RXR) are key regulators in moulting. In the present study, the full-length cDNAs of the MIH, EcR2, and RXR3 genes from the red swamp crayfish, Procambarus clarkii (Girard, 1852) (denoted as PcMIH, PcEcR2, and PcRXR3) were cloned. Tissue-specific and moult stage-specific mRNA expression patterns of these genes were detected by real-time quantitative polymerase chain reaction. PcMIH was detected only in the eyestalk, whereas PcEcR2 and PcRXR3 mRNA were expressed in all tissues tested. The highest levels of PcEcR2 and PcRXR3 were detected in the gill and hepatopancreas. Expression of PcMIH mRNA in the eyestalk increased from postmoult to peak in intermoult and then decreased in premoult. Expression of PcEcR2 mRNA in the eyestalk, hepatopancreas, and muscle increased from postmoult to peak in early premoult and then decreased. However, expression of PcEcR2 mRNA in the gill increased from postmoult to reach a maximum in intermoult and then decreased in premoult. Expression of PcRXR3 mRNA also fluctuated in the eyestalk, hepatopancreas, muscle, and gill, with a decrease from postmoult to late premoult. Expression of PcEcR2 and PcRXR3 mRNA increased relative to the control in the hepatopancreas and gill after unilateral and bilateral eyestalk ablation, which suggested that PcMIH can inhibit their mRNA expression. Double-stranded RNA-mediated RNA interference of PcRXR3 caused different changes in mRNA expression of these genes in different tissues and resulted in decreased expression of PcEcR2 mRNA, which suggested a collaborative relationship between PcEcR2 and PcRXR3.
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