ABSTRACT
Crab grows by periodic molting, which is controlled by molt-inhibiting hormone (MIH) and ecdysteroids. Untranslated regions (UTRs) play crucial roles in the posttranscriptional regulation of gene expression. In this study, using crab collected from Changjiang (Yangtze), Huanghe (Yellow), Liaohe, and Yalujiang rivers, 33 haplotypes of the 3ê-UTR of ecdysteroid-regulated protein (ERP) gene were identified, of which 14 haplotypes were observed in more than one individual. One hundred and forty clones of haplotype h2 (41.5%) were observed in samples from all the 4 rivers. Three haplotypes were observed to be insertions. For the whole crab sample, we found a positive Tajima's D value and a negative Fu's Fs test (Tajima's D value = 0.98726; Fu's Fs test = -27.382), although the P values were not significant (P > 0.10). The network profile of these 33 haplotypes presented a single core pattern with h2 as the core. In this study, we found that the UTR of ERP gene had a considerably high genetic polymorphism among crab from regions south to north of China. Furthermore, we observed a relatively high genetic divergence among different haplotypes, which would suggest a high diversity of the crab gene pool.
Subject(s)
Brachyura/genetics , Genetic Variation/genetics , Molting/genetics , 3' Untranslated Regions/genetics , Animals , Haplotypes/genetics , Polymorphism, Genetic/geneticsABSTRACT
We explored the protective effect of ischemia preconditioning (IP) on ischemia-reperfusion injury in rat liver transplantation. An orthotopic liver transplantation model was utilized in the study. A total of 54 Sprague-Dawley rats were divided into a control group (group A, no liver transplantation), liver transplantation group (group B, heparin Ringer's lactate solution was perfused via the portal vein before donor liver collection), and liver transplantation with IP group (group C, IP was performed for different time periods before donor liver collection). Liver function, B-cell lymphoma 2 expression in hepatic cells, cell apoptosis, and cellular ultrastructure changes were detected after surgery. After surgery, serum alanine aminotransferase activity was significantly higher in group B than in group A, while it was not clearly enhanced in group C and decreased progressively with increasing cycles of IP as bile capacity gradually increased. Compared with group B, group C showed alleviated injury of hepatic cells, increased B-cell lymphoma 2 expression, and a lower apoptosis index. IP had a protective effect on ischemia-reperfusion injury in rat liver transplantation, and the mechanism correlated with increased B-cell lymphoma 2 expression in hepatic cells and inhibition of cell apoptosis.
Subject(s)
Ischemic Preconditioning/methods , Liver Transplantation/methods , Reperfusion Injury/metabolism , Reperfusion Injury/therapy , Animals , Case-Control Studies , Liver/metabolism , Liver/pathology , Liver Function Tests , Male , Proto-Oncogene Proteins c-bcl-2/metabolism , Random Allocation , Rats , Rats, Sprague-DawleyABSTRACT
C-type lectins are a superfamily of Ca(2+)-dependent carbohydrate-recognition proteins that are well known for their participation in pathogen recognition and clearance. In this study, a putative C-type lectin fold (MyCLF) gene was identified from the Japanese scallop Mizuhopecten yessoensis. The full-length of MyCLF was 645 bp, encoding a polypeptide of 167 amino acids. MyCLF carried a signal peptide of 20 amino acid residues, and a single carbohydrate recognition domain, having relatively high amino acid sequence conservation with C-type lectins reported for other bivalves. The expression of MyCLF mRNA transcripts in adult tissues, after bacterial challenge and during different developmental stages was determined using real-time quantitative RT-PCR. MyCLF was mainly distributed in the mantle, gill, and kidney. The expression of MyCLF clearly increased 3 h after Vibrio anguillarum challenge, and dropped to a minimum level after 9 h compared to the control group. During embryonic development, the expression level increased in the gastrulae, trochophore and early D-shaped larvae, decreased in D-shaped larvae, and then increased hundreds of times in metamorphosing larvae. The results suggested that MyCLF was involved in an immune response and it may play important roles during the metamorphosis phase of M. yessoensis.
Subject(s)
Immunity/genetics , Lectins, C-Type/genetics , Metamorphosis, Biological/genetics , Pectinidae/genetics , Amino Acid Sequence , Animals , Base Sequence , Cloning, Molecular , DNA, Complementary/chemistry , DNA, Complementary/genetics , Gene Expression Profiling , Gene Expression Regulation, Developmental , Larva/genetics , Larva/growth & development , Lectins, C-Type/chemistry , Lectins, C-Type/classification , Models, Molecular , Molecular Sequence Data , Pectinidae/embryology , Pectinidae/growth & development , Phylogeny , Protein Conformation , Reverse Transcriptase Polymerase Chain Reaction , Sequence Analysis, DNA , Sequence Homology, Amino AcidABSTRACT
Chitinase-like proteins (CLP) are important members of the glycoside hydrolase family 18 (GH18) and are involved in growth control and remodeling processes. In this study, a CLP transcript was isolated and sequenced from the Japanese scallop (Mizuhopecten yessoensis) after screening expressed sequence tags. The full-length complementary DNA of M. yessoensis CLP (My-Clp1) was 1555 bp in length, consisting of a 75-bp 5'-untranslated region (UTR), a 160-bp 3'-UTR, and a 1320-bp open reading frame bearing characteristics of the GH18 family. The My-Clp1 protein was well conserved, with similar domain structures and architecture across species (e.g., from mollusks to mammals). Expression analysis in healthy tissues and across developmental stages revealed a strong preference for expression; My-Clp1 was abundantly expressed in the mantle and throughout metamorphosis, which suggests the involvement of My-Clp1 in the synthesis of extracellular components, and tissue degeneration and remodeling. My-Clp1 expression was induced after infection with a bacterial pathogen, Vibrio anguillarum, suggesting its involvement in immunity against this intracellular pathogen.
Subject(s)
Chitinases/genetics , DNA, Complementary/genetics , Pectinidae/physiology , Transcriptome/physiology , Amino Acid Sequence , Animals , Base Sequence , Chitinases/chemistry , Chitinases/metabolism , DNA, Complementary/metabolism , Molecular Sequence Data , Organ Specificity/genetics , Pectinidae/microbiology , Phylogeny , Vibrio/pathogenicity , Vibrio Infections/metabolismABSTRACT
Japanese scallop (Mizuhopecten yessoensis) is a cold-water shellfish, and a species of economic importance in China. In this study, we developed and evaluated simple sequence repeat (SSR) markers from the expressed sequence tags (ESTs) of M. yessoensis. The characteristics of 12 EST-SSR loci were investigated in 30 individual scallops, and the result revealed that the number of alleles per locus ranged from 2-4, with an observed heterozygosity ranging from 0.0333-0.7692, and an expected heterozygosity ranging from 0.0333-0.6312. Only two loci were found to depart significantly from the Hardy-Weinberg equilibrium (P < 0.05). The result of our study suggested that these markers could be considered as potential markers for studying the population structure of M. yessoensis and its intraspecific variation.