Characterization of hemocyanin from the mud crab Scylla paramamosain and its expression analysis in different tissues, at various stages, and under Vibrio parahaemolyticus infection

Abstract

ABSTRACT.Hemocyanin is an important respiratory protein in many arthropod and mollusk species. Here, four cDNAs (SpHc1, SpHc2, SpHc3, and SpHc4), encoding distinct hemocyanin subunits from Scylla paramamosain were cloned using EST analyses and the rapid amplification of cDNA ends. The four full-length cDNA fragments (SpHc1-4) were 2281, 2002, 2184, and 2069 bp, respectively, and they encoded four putative proteins (570-676 amino acids) with a molecular mass of ~65.0-76.8 kDa. Quantitative real-time PCR analyses revealed that the four genes were mainly expressed in the hepatopancreas, testis, and hemocytes. SpHc mRNA expression during continuous developmental stages in zoeal phases (Z1, Z2, Z3, Z4, and Z5), megalopa, and juvenile crab I stages were also detected. The expression levels of SpHc3 and SpHc4 were higher than that of SpHc1 and SpHc2 during the first six stages, and they sharply declined during the juvenile stage. After infection with Vibrio parahaemolyticus, the temporal expression of both the four SpHc mRNAs in the megalopa stage rapidly declined during the first 3 h, followed by upregulation and peak expression at 12 h after the challenge. The expression levels of the four SpHc subunits were upregulated at 48 h after the challenge, and were then gradually downregulated. These findings suggest that hemocyanin may potentially be involved in the crab immune response, and that the role of the four subunits may differ in different tissues and during various developmental stages.

Prediction of Scylla olivacea (Crustacea; Brachyura) peptide hormones using publicly accessible transcriptome shotgun assembly (TSA) sequences

“…As part of the ongoing effort to support and enhance the aquaculture of Scylla species, much work has focused on understanding the biology of these crabs (e.g., Bao et al, 2015;Chen et al, 2015;Girish et al, 2015;Gong et al, 2015;Huang et al, 2014Huang et al, , 2015Ikhwanuddin et al, 2014;Kornthong et al, 2014;Sun et al, 2015;Wang et al, 2015aWang et al, , 2015bXie et al, 2014;Zeng et al, 2016;Zhao et al, 2015). One outcome of these studies has been the development of extensive transcriptomic resources for several Scylla species (e.g., Bao et al, 2015;Kornthong et al, 2014;Ma et al, 2014;Xie et al, 2014), including S. olivacea, for which approximately 160,000 transcriptome shotgun assembly (TSA) sequences are now available in GenBank (e.g., BioProject Accession No.…”

Antibacterial activity of crab haemocyanin against clinical pathogens

“…Similarly, horse crab Portunus trituberculatus and mangrove crab Scylla serrata reportedly have five types of monomeric arthropod haemocyanin subunits [36,37]. Some crustaceans express distinct haemocyanin subunits during certain developmental stages, or synthesise such subunits in response to certain physiological conditions [38]. Many researchers have reported that the expression levels of hemocyanin genes are affected by bacterial or viral infection [30,38].…”

“…Some crustaceans express distinct haemocyanin subunits during certain developmental stages, or synthesise such subunits in response to certain physiological conditions [38]. Many researchers have reported that the expression levels of hemocyanin genes are affected by bacterial or viral infection [30,38]. These data could explain the presence of a different number of glycosylated haemocyanin subunits in the haemolymph of E. verrucosa.…”