Crucial roles of a novel exoskeletal-derived lectin in innate immunity of the oriental river prawn, Macrobrachium nipponense.

As important pattern recognition receptors (PRRs), C-type lectins play crucial roles in the crustacean innate immune system. In this study, a novel C-type lectin, designated as MnLec1, was obtained from the exoskeleton of the oriental river prawn Macrobrachium nipponense for the first time. The full-length cDNA of MnLec1 was 1329 bp with an open reading frame of 774 bp. The predicted MnLec1 protein contains a single carbohydrate-recognition domain with an EPN/LND motif and one Ca2+ binding site-2. MnLec1 transcripts were widely detected in the tested tissues of M. nipponense and significantly up-regulated after Aeromonas hydrophila challenge. The recombinant MnLec1 protein was found to have a wide spectrum of binding activities towards various microorganisms, agglutinate two kinds of Gram-negative bacteria (Escherichia coli and A. hydrophila) in a Ca2+ -independent manner. What's more, the survivability of prawns was significantly down-regulated after RNAi of MnLec1 when infected with A. hydrophila. Collectively, these findings suggest that MnLec1 from the exoskeleton might function as a PRR and play a crucial role in immune defense against invading pathogens in M. nipponense.

Reconstruction and analysis of a genome-scale metabolic model for Eriocheir sinensis eyestalks.

The eyestalk of Eriocheir sinensis has significant biological functions with many nerve peptide hormones expressed in the X-organ which exists in the eyestalk. A metabolic network model is an effective tool for the systematic study of E. sinensis eyestalks. In this work, we reconstructed a metabolic network model for E. sinensis eyestalks based on transcriptome sequencing. The model contains 1304 reactions, 1381 unigenes and 1243 metabolites distributing in 98 pathways. The reconstructed metabolic network model was used for the functional module and block metabolite analysis of eyestalks, which reveals that the function of the eyestalk network agrees with its function as the centre of the E. sinensis endocrine system. The difference expression analysis of reactions in the model indicates that the eyestalk mainly functions in the regulation of amino acids, carbohydrate and nucleotide metabolism.

A novel effect of imidazole derivative KK-42 on increasing survival of Aeromonas hydrophila challenged prawn Macrobrachium nipponense.

Imidazole derivative KK-42 is well known as the insect growth regulator. Here we find that KK-42 pretreatment could promote the survival of Macrobrachium nipponense infected with Aeromonas hydrophila, which is considered to be possibly related to the prophenoloxidase (proPO), a conserved copper-containing enzyme that plays an important role in defense against pathogens. In this study, a full-length of proPO gene from M. nipponense haemocytes, designated as MnproPO, was firstly cloned and characterized. The full-length cDNA contained 2428 bp with a 2013 bp open reading frame encoding a putative proPO protein of 671 amino acids with a predicted molecular mass of 76.5 kDa and pI of 7.31. It was predicted to possess all the expected features of proPO members, including two putative copper-binding sites with six histidine residues and a thiol ester-like motif. Sequence analysis showed that MnproPO exhibited the highest amino acid sequence similarity (93%) to a proPO of Macrobrachium rosenbergii. The gene was expressed highly in haemocytes and weakly in hepatopancreas. Real-time PCR analysis revealed that the MnproPO expression increased significantly at 3, 12 and 24 h after KK-42 treatment, the PO activity also importantly rose from 6 to 48 h in KK-42-treated prawns and reached the maximum at 24 h with a 2.3-fold higher than that in control group. Injection of A. hydrophila could stimulate the MnproPO transcription and PO activity whether or not the prawns were pretreated by KK-42, the mRNA level increased obviously only at 3 h and 6 h after the bacterium injection (challenged control), but increased constantly during the phase of experiment except at 6 h under the condition of KK-42 pretreatment (challenged treatment group). The change trend of PO activity was basically similar to that of MnproPO expression. Our present results demonstrate that the MnproPO expression as well as PO activity may be induced by KK-42, which is likely one of the molecular mechanisms of KK-42 acts for increasing survival of the prawn infected with A. hydrophila.

Research for the effect of octylphenol on spermatogenesis and proteomic analysis in octylphenol-treated mice testes.

OP (octylphenol), an environmental oestrogen was administered, and differentially expressed proteins were analysed in mice testes to clarify its mechanism of action in male sterility. Male Kunming suckling mice (10 days old) were subcutaneously injected with OP at a dose of 10 µg/kg per day, 50 µg/kg per day and 100 µg/kg per day as low-, medium- and high-dose groups, respectively, for 35 days. Animals in the control group received subcutaneous injections of olive oil at a dose of 10 µl/mouse per day. Serum oestradiol, testosterone, FSH (follicle stimulating hormone) and LH (luteinizing hormone) levels were measured on day 45. The left testes were removed for tissue analysis, and the right testes were analysed for differentially expressed proteins by using two-dimensional gel electrophoresis and MS. Tissue analysis showed that mice spermatogenesis was blocked at the round spermatid stage in the high-dose group, whereas no such changes were found in the medium- and low-dose groups. Higher serum oestradiol (P<0.05) and lower testosterone (P<0.05) levels were found in the medium- and high-dose groups. There was no significant difference in serum oestradiol and testosterone levels in the low-dose and control groups. No significant influence of OP was seen on serum FSH and LH levels in all OP-treated animals. The results from four differentially expressed proteins such as PPIA (peptidyl-prolyl cis-trans isomerase A), PEBP1 (phosphatidylethanolamine-binding protein1), TPI (triose-phosphate isomerase) and TCP-1 (T-complex protein 1) in the high-dose and control groups showed up-regulation of PPIA expression and down-regulation in PEBP1, TPI and TCP-1 expressions. These findings will contribute to clarify the mechanism of male sterility by environmental oestrogens.

Regulatory effects of corticosterone on ornithine decarboxylase activity during liver regeneration in rats.

AIMS: The regulation of ornithine decarboxylase (ODC) gene expression and enzyme activity by corticosterone during rat liver regeneration induced by partial hepatectomy (PH) was evaluated. METHODS: Bilateral adrenalectomies were performed on ether-anesthetized rats 3 days before PH. Corticosterone in sesame oil was injected subcutaneously to adrenalectomized rats. ODC mRNA, ODC protein and enzyme activity were detected by in situ hybridization, Western blot and high performance liquid chromatography (HPLC), respectively. RESULTS: The ODC mRNA levels, protein accumulation and enzyme activity were lower in the intact liver compared to the regenerating liver. After PH, mRNA levels were remarkably enhanced in all groups and peaked at 5 h post-PH, and presented a persistent increase only in adrenalectomy rats during the regeneration process. Corticosterone treatment resulted in a dose-dependent decrease in ODC mRNA content after 5 h post-PH. ODC protein accumulation in adrenalectomy rats was higher than that in sham-adrenalectomy rats, but it decreased in corticosterone-treated (10 mg/kg) rats until 24 h post-PH, with a strong decline seen in 40 mg/kg corticosterone-treated rats. ODC activity was rapidly promoted, and the highest levels were observed at 6 h after PH in all groups. After corticosterone treatment, the activities declined significantly at 6 h post-PH, with the lowest value found in the 40 mg/kg group. CONCLUSIONS: Corticosterone treatment results in dose-dependent decreases in ODC mRNA and enzyme protein both in the intact liver and the regenerating liver. The change in ODC activity is partially related to alterations of ODC mRNA and protein accumulation.

[Progress in the research of antizyme].

Antizyme is a small protein induced by elevated intracellular polyamines. Antizyme binds specifically to ornithine decarboxylase (ODC) , targeting ODC for ubiquitin-independent degradation by the 26S protosome, which consequently reduces polyamine synthesis. Polyamine transport may also be regulated by antizyme, through which the cellular polyamine levels are maintained. In recent years, more and more studies focus on antizyme with the development of biotechnique. In this article, advancements in the antizyme family, synthesis, function and intracellular location are reviewed.

Expression patterns and action analysis of genes associated with drug-induced liver diseases during rat liver regeneration.

AIM: To study the action of the genes associated with drug-induced liver diseases at the gene transcriptional level during liver regeneration (LR) in rats. METHODS: The genes associated with drug-induced liver diseases were obtained by collecting the data from databases and literature, and the gene expression changes in the regenerating liver were checked by the Rat Genome 230 2.0 array. RESULTS: The initial and total expression numbers of genes occurring in phases of 0.5-4 h after partial hepatectomy (PH), 4-6 h after PH (G0/G1 transition), 6-66 h after PH (cell proliferation), 66-168 h after PH (cell differentiation and structure-function reconstruction) were 21, 3, 9, 2 and 21, 9, 19, 18, respectively. It is illustrated that the associated genes were mainly triggered at the initial stage of LR and worked at different phases. According to their expression similarity, these genes were classified into 5 types: only up-regulated (12 genes), predominantly up-regulated (4 genes), only down-regulated (11 genes), predominantly down-regulated (3 genes), and approximately up-/down-regulated (2 genes). The total times of their up- and down-expression were 130 and 79, respectively, demonstrating that expression of most of the genes was increased during LR, while a few decreased. The cell physiological and biochemical activities during LR were staggered according to the time relevance and were diverse and complicated in gene expression patterns. CONCLUSION: Drug metabolic capacity in regenerating liver was enhanced. Thirty-two genes play important roles during liver regeneration in rats.