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Electron. j. biotechnol ; 19(6): 49-55, Nov. 2016. ilus
Article in English | LILACS | ID: biblio-840313


Background: Bisphenol A (BPA) is an endocrine-disrupting chemical (EDC) with a weak estrogen-like activity in fish that is found ubiquitously in aquatic environments. However, there has been little study about BPA on the endocrine disrupting effects of crab. In the present study, cDNA of vasa was cloned and characterized in the Charybdis japonica. Histological structures of testis and expression patterns of vasa gene in the testis of C. japonica after treatment with BPA were investigated. Results: The cDNA of vasa is composed of 3051 bp with a 2166 bp open reading frame encoding 721 AA. The deduced amino acid sequence contained eight conserved domains of the DEAD-box protein family. The tissue distribution showed that vasa mRNA was specifically expressed in ovary and testis. Histologically, the sperm cells were decreased in number and an acellular zone was seen in the testis. The transcript level of vasa gradually increased with a significant difference between the experimental and control groups. After BPA exposure with 0.50 and 1.00 mg/L for 1,3, 6 and 9 d, the expression levels of vasa increased. Conclusion: These findings suggest that BPA can increase the expression level of vasa mRNA and influence the development of the testis in C. japonica.

Animals , Male , Benzhydryl Compounds/pharmacology , Brachyura/drug effects , Brachyura/genetics , DEAD-box RNA Helicases/drug effects , DEAD-box RNA Helicases/genetics , Phenols/pharmacology , Cloning, Molecular , Endocrine System/drug effects , Nucleic Acid Amplification Techniques , Real-Time Polymerase Chain Reaction , Testis/drug effects
Electron. j. biotechnol ; 16(2): 2-2, Mar. 2013. ilus, tab
Article in English | LILACS | ID: lil-670130


Background: Cyclophilin A (CypA), a receptor for the immunosuppressive agent cyclosporin A (CsA), is a cis-trans peptidyl-prolyl isomerase (PPIase) which accelerates the cis-trans isomerization of prolyl-peptide bonds and interacts with a variety of proteins to regulate their activities. Results: The full-length cDNA of crab Eriocheir sinensis CypA (EsCypA) was cloned by EST and RACE technique. The complete sequence of EsCypA cDNA contained a 5' untranslated region (UTR) of 50 bp, a 3’ UTR of 233 bp with a polyA tail, and an open reading frame (ORF) of 495 bp encoding a polypeptide of 164 amino acids with the predicted molecular weight of 17.36 kDa. The deduced amino acid sequence of EsCypA contained two highly conserved signature sequences of peptidyl-prolyl cis-trans isomerase and a pro-isomerase domain. The mRNA transcripts of EsCypA were detectable in all the examined tissues, including haemocytes, gill, hepatopancreas, gonad, muscle and heart, with higher expression level in hepatopancreas and gonad. No significant difference in the relative mRNA expression level of EsCypA was observed during the whole course of bacteria challenge, whereas it was up-regulated during fungi challenge. The purified recombinant protein rEsCypA exhibited a significant PPIase activity and an antifungal activity. Conclusions: All these results indicated that it was a typical CypA member and potentially involved in the innate immune responses of crab.

Animals , Brachyura/genetics , Brachyura/immunology , Cyclophilin A , Antifungal Agents , Phylogeny , Sequence Alignment , Sequence Analysis , Immunity
Genet. mol. biol ; 34(3): 529-531, 2011. ilus
Article in English | LILACS | ID: lil-595991


The decapod Grapsus grapsus is commonly found on oceanic islands of the Pacific and Atlantic coasts of the Americas. In this study, a simple, quick and reliable method for detecting its larvae in plankton samples is described, which makes it ideal for large-scale studies of larval dispersal patterns in the species.

Animals , Brachyura/genetics , Genetic Markers , Larva/genetics , Plankton/genetics
Genet. mol. biol ; 34(1): 136-141, 2011. graf, tab
Article in English | LILACS | ID: lil-573719


The identification of expressed genes involved in sexual precocity of the mitten crab (Eriocheir sinensis) is critical for a better understanding of its reproductive development. To this end, we constructed a cDNA library from the rapid developmental stage of testis of E. sinensis and sequenced 3,388 randomly picked clones. After processing, 2,990 high-quality expressed sequence tags (ESTs) were clustered into 2,415 unigenes including 307 contigs and 2,108 singlets, which were then compared to the NCBI non-redundant (nr) protein and nucleotide (nt) database for annotation with Blastx and Blastn, respectively. After further analysis, 922 unigenes were obtained with concrete annotations and 30 unigenes were found to have functions possibly related to the process of reproduction in male crabs - six transcripts relevant to spermatogenesis (especially Cyclin K and RecA homolog DMC1), two transcripts involved in nuclear protein transformation, two heat-shock protein genes, eleven transcription factor genes (a series of zinc-finger proteins), and nine cytoskeleton protein-related genes. Our results, besides providing valuable information related to crustacean reproduction, can also serve as a base for future studies of reproductive and developmental biology.

Animals , Male , Brachyura/genetics , Gene Expression Profiling , Reproduction , China , DNA, Complementary , Expressed Sequence Tags