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1.
Raptor, a member of the target of rapamycin complex 1 (TORC1), participates in the formation of complex proteins related to the mechanistic target of rapamycin (mTOR) signalling. In this study, a 5,020 bp cDNA of Raptor with an open reading frame (ORF) of 3,804 bp encoding for 1,267 amino acids was cloned from Litopenaeus vannamei. The protein contains three conserved domains: Raptor N, HEAT and WD40 domains. The expression of Raptor gene was detected by qRT‐PCR in different tissues of L. vannamei, including hepatopancreas, intestinal, stomach, eyestalk, gill and muscle. The mRNA expression profiles of Raptor in muscle were also analysed under suppression or stimulation of mTOR signalling pathway. The level of Raptor mRNA significantly increased either at 0.5–6 hr after an injection of rapamycin (RAPA) or after 3 days starvation. Leucine or arginine alleviated the up‐regulation of Raptor gene expression caused by RAPA or starvation. The Raptor gene was successfully suppressed using RNA interference (RNAi) technology, and the gene expression and the protein phosphorylation level of 4EBP1 and S6K were significantly decreased. The results of the study suggested that the expression of Raptor was sensitive to the immunology status of L. vannamei and participated in nutritional metabolism.  相似文献   

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3.
The present study was to determine the time‐dependent alterations in growth performance, histomorphology, digestive enzymes activities and the genes expression related to target of rapamycin (TOR) signalling pathway involved in hepatopancreas and intestine of Litopenaeus vannamei at 3, 6, 12, 18, 24 and 30 days after Aflatoxin B1 (AFB1) challenge. The results manifested that AFB1 could induce a significant decrease in growth performance and visible variations of the hepatopancreas and intestine structures in shrimp. Meanwhile, the digestive enzymes activities and genes expression level were significantly lower in the experimental group than in the control group and showed a time‐dependent decrease (p < 0.05). The expression levels of tor and s6k were significantly higher in the experimental group than in the control group from days 6 to 24 and showed a tendency to increase first and then decrease as a whole (p < 0.05). Meanwhile, we analysed candidate pathways involved in hepatopancreas and intestine of shrimp in metabolic response to AFB1. The mainly disturbed pathways related to metabolism in hepatopancreas were involved in pyrimidine, glycosylphosphatidylinositol, glycosaminoglycan, vitamin and amino acid biosynthesis and metabolism, while the mainly disturbed metabolic pathways in intestine were glycosaminoglycan, vitamin and amino acid biosynthesis and metabolism.  相似文献   

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5.
The complement component 9 (C9) plays a significant role in the formation of membrane attack complex (MAC) on the targeted cell surface. The current study is dealt with molecular characterization of C9 gene from rohu, Labeo rohita, an important cultured carp species in India. An open reading frame (ORF) of 1998 bp was amplified by polymerase chain reaction (PCR) that encodes a polypeptide of 666 amino acids having a signal peptide of 19 amino acids and a mature peptide of 647 amino acids. The SMART domain architecture analysis revealed two thrombospondin type‐1 domains (TSP1), a low‐density lipoprotein receptor domain class A (LDLa), a membrane attack complex and perforin (MACPF) domain, and an epidermal growth factor (EGF)‐like domain. Multiple sequence alignment and evolutionary analysis revealed a primitive C9 sequence of rohu with maximum similarity and clustering with common carp, grass carp and zebrafish. C9 was highly expressed in liver and constitutively expressed in wide array of tissues except in eye of rohu juveniles. It was expressed during early developmental days of rohu including in milt. A variable level of up‐regulation in C9 expression was noticed upon poly I:C induction, Aeromonas hydrophila and Argulus siamensis infections in liver, spleen and gill tissues of rohu at different time points. A constitutive expression of C9 in different stages of rohu during the ontogeny and in response to pathogen exposures along with high degree of sequence homology with other fish species proved it as an important primitive immune molecule of the complement system lytic pathway.  相似文献   

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7.
Toll‐like receptors (TLRs) play an indispensable role in fish immunity, being involved in pathogen recognition and the triggering of immune reactions. Here, a member of the TLR family, TLR1, from Lateolabrax japonicus was characterized and its expression pattern and intracellular localization were analysed. The full‐length LjTLR1 cDNA (2,755 bp) was found to encode a polypeptide of 827 amino acids. The deduced amino acid sequence contained three main structural domains: an extracellular leucine‐rich repeat domain, a transmembrane domain and a Toll/IL‐1 receptor domain. Tissue distribution analysis indicated that LjTLR1 was expressed in all of the examined tissues to varying degrees, with the highest levels being measured in the head kidney. In order to assess the antibacterial functions of LjTLR1 during infection, the pathogenic bacteria Vibrio harveyi and Streptococcus agalactiae were used. LjTLR1 was significantly upregulated in the three immune organs (the head kidney, spleen and liver) following bacterial stimulation, and its expression was detected 6 hr after initial exposure. In mRNA in situ hybridization experiments, positive signals were more numerous in the treatment group than the control group, verifying the expression patterns observed. Assessment of the intracellular localization of LjTLR1 revealed it to be present in the cytoplasm. These results indicate the potential role of LjTLR1 in immune responses to bacterial infection. This study enriches our knowledge of L. japonicus immune genes and provides a theoretical basis for further research concerning the antibacterial functions of fish TLRs during infection.  相似文献   

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10.
The use of traditional bait and medicament in freshwater aquaculture exacerbates environmental pollution and leads to frequent occurrence of diseases. Effluent collected after Rhodopseudomonas palustris‐mediated wastewater treatment could be reutilized as microbial feeds, medicament and aquaculture water to culture Pelteobagrus vachelli. Therefore, a novel integrated system of wastewater treatment using effluent containing R. palustris that improves yield, increases disease resistance and enhances the quality of aquaculture water for P. vachelli culture was proposed and investigated. P. vachelli can grow well in effluent containing R. palustris (ER). The survival rate, yield and whole‐body composition of the ER group were all increased compared to those of the control group (CK). The biochemical (B vitamin) and other substances in the effluent of R. palustris enhanced the activity of proteases, amylases, lipases, alkaline phosphatase (AKP), acid phosphatase (ACP), phagocytic, superoxide dismutase (SOD) and catalase (CAT) by up‐regulating the expression of AKP, ACP, SOD and CAT genes. Theoretical analysis showed that biochemical substances regulated the expression of these genes and enzyme activities as stimulus signal, component and active centre. Moreover, R. palustris and biochemical substances improved the target of rapamycin (TOR) and nuclear factor kappa B (NF‐κB) signalling pathways and intestinal microbiota. Furthermore, R. palustris inhibited Aeromonas hydrophila, which increased resistance to fish diseases and promoted the growth of intestinal probiotics. Meanwhile, R. palustris in effluent also purified the quality of aquaculture water. Use of this technology simultaneously helped improve aquaculture water quality, reduce water pollution and wastewater discharge and increased the output and disease resistance of P. vachelli.  相似文献   

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12.
A 9‐week feeding trial was conducted to estimate the dietary isoleucine requirement of juvenile blunt snout bream. Six isonitrogenous and isoenergetic experimental diets were formulated to contain graded isoleucine levels ranging from 5.3 to 20.1 g kg?1 dry diet. At the end of the experiment, weight gain (WG), specific growth rate (SGR), feed efficiency ratio (FER) and protein efficiency ratio (PER) increased with increasing dietary isoleucine level up to 11.1 g kg?1 dry diet, and dietary isoleucine level above 14.2 g kg?1 dry diet declined these performances. Dietary isoleucine levels (14.2 and 17.3 g kg?1 dry diet) significantly improved whole‐body protein content, but decreased whole‐body lipid, plasma triglyceride and cholesterol contents. Significantly lower visceral fat index (VFI) in fish fed with 14.2 g kg?1 dietary isoleucine was observed compared to those fed with deficient or excessive isoleucine. Dietary isoleucine supplementation significantly increased plasma isoleucine concentration, while plasma valine and leucine concentrations showed a reversed trend. Dietary isoleucine levels regulated the target of rapamycin (TOR) gene expression and improved plasma superoxide dismutase (SOD) activity in juvenile blunt snout bream. Based on second‐order polynomial regression model analysis of SGR and FER, the optimum dietary isoleucine requirement was estimated to be 13.8 g kg?1 dry diet (40.6 g kg?1 dietary protein) and 14.0 g kg?1 dry diet (41.2 g kg?1 dietary protein), respectively.  相似文献   

13.
Six diets were formulated to investigate the success of fish meal (FM) replacement by plant proteins; diet 1 reflected a commercial feed (8% FM), diet 3 contained 4% FM, and diet 5 was devoid of FM. Whereas, diet 2, diet 4, and diet 6 reflected diet 1, diet 3, diet 5, respectively, and supplemented with essential amino acid (EAA). At the end of 8‐week trial, there was no significant difference in survival rate. Significantly higher final weight, weight gain rate, and specific growth rate were recorded in the group fed diet 2 compared with the other treatments (except diet 4) (p < 0.05). Feed conversion ratio of fish fed diet 2 was significantly lower than those fed diets 1, 3, and 5 (p < 0.05). The lowest feed intake and highest protein efficiency rate were found in fish fed diet 2 (p < 0.05). There was no significant difference in whole body compositions between treatments. Plasma aspartate transaminases, alanine aminotransferase, and glucose were significantly affected by dietary treatments (p < 0.05), while plasma protein and albumin contents were not influenced by the treatments. The relative expression of target of rapamycin (TOR) and phosphatidylinositol 3‐kinase, regulatory subunit 1 (alpha) (PIK3R1) in fish fed diet 3 (4% FM) were significantly down‐regulated compared with those fed diet 6 for TOR and diets 4 and 6 for PIK3R1 (p < 0.05). Insulin receptor substrate 1 (IRS‐1) and janus kinase 3 (JAK3) expressions were fluctuated, with the higher levels in fish fed diets 4 and 6. In conclusion, the findings of this study indicate that plant protein mixture supplemented with EAA could be used to substitute FM in practical diet for Megalobrama amblycephala.  相似文献   

14.
One hundred and twenty crabs (initial weight: 51.32 ± 0.06 g) were fed three experimental diets with cottonseed meal protein hydrolysate (CPH) at 0, 6 and 12 g/kg for 60 days. The results showed that CPH0.6 diet significantly improved the 1‐hr feed intake, protease activity and weight gain of crabs. In thoracic ganglia, 4E‐binding protein (4E‐BP) expression level of crab fed the diets containing CPH was significantly higher than CPH0 group. In cerebral ganglia, protein kinase B and 4E‐BP expression levels of crab fed CPH0.6 diets were significantly lower than CPH0 group. In mid‐intestine, target of rapamycin (TOR) relative expression level of crabs fed the diets containing CPH was significantly lower than CPH0 group. 4E‐BP expression level showed an opposite pattern. In thoracic ganglia, cerebral ganglia and mid‐intestine, leptin receptor expression level of crabs fed CPH0.6 diet was significantly lower than CPH0 group. Neuropeptide Y and neuropeptide Y receptor expression level of crab fed CPH0.6 diet was significantly higher than CPH0 group. In conclusion, our results indicated that dietary supplementation of 6 g/kg CPH could stimulate appetite and quicken feeding rate via the TOR signalling pathway.  相似文献   

15.
The γ‐aminobutyrate type A receptor‐associated protein (GABARAP) is a ubiquitin‐like modifier implicated in membrane trafficking and fusion events involving the γ‐aminobutyrate type A receptor, autophagy and apoptosis. In this study, the gene encoding GABARAP was cloned from swimming crab Portunus trituberculatus (PtGABARAP) based on the expression sequence tag (EST). The full‐length cDNA of 664 bp includes a 5′ untranslated region (UTR) of 87 bp, a 3′ UTR of 223 bp with a poly(A) tail, and an open reading frame (ORF) of 354 bp encoding a polypeptide of 117 amino acids with a predicted molecular weight of 13.96 kDa. The deduced amino acid sequence shares high similarity (93%–100%) with GABARAPs from other species and includes a conserved Atg8 domain. In a phylogenetic analysis PtGABARAP clustered with GABARAPs from other species, and more widely with other GABARAP family proteins. The impact of elevated ocean acidification (OA) on P. trituberculatus behaviours was investigated, and real‐time RT‐PCR revealed that PtGABARAP expression was up‐regulated after OA exposure. Ocean acidification also caused crabs anxiety‐like behaviours, like the shoal average speed increase, preference for dark environment (scototaxis) and fast exploration. The results indicated that GABARAP might be involved in the interactions of GABAA receptors and elevated‐CO2 seawater.  相似文献   

16.
The experiment was conducted to determine the leucine requirement of juvenile Pacific white shrimp Litopenaeus vannamei (Boone) in low‐salinity water (0.50–1.20 g L?1). Six diets were formulated to contain 410 g kg?1 crude protein with fish meal, peanut meal and precoated crystalline amino acids with different concentration of l ‐leucine (16.72, 19.60, 22.06, 24.79, 27.28 and 30.16 g kg?1 dry diet). Each diet was randomly assigned to triplicate groups of 30 shrimps (0.38 ± 0.002 g), and the feed trial lasted for 8 weeks. The results indicated that the maximum weight gain was observed at 24.95 g kg?1 dietary leucine group, whereas the diets containing higher leucine concentration conversely reduced the growth performance (P < 0.05). Moreover, the highest body protein content and body protein deposition and the lowest haemolymph AST and ALT activities were also found at 24.95 g kg?1 dietary leucine group. With the increase in leucine in diets, a dose‐dependent increase was found in body lipid content and haemolymph urea concentration. The polynomial regression calculated using weight gain, feed efficiency and body protein deposition indicated that the optimal dietary leucine requirement for L. vannamei reared in low‐salinity water was 23.73 g kg?1 leucine of dry diet, correspondingly 57.88 g kg?1 of dietary protein.  相似文献   

17.
O‐GlcNAc transferase gene (OGT) was considered as the sole rate‐limiting enzyme in the O‐GlcNAc modification. In the present study, the OGT gene of hybrid grouper (Epinephelus fuscoguttatus ♀ × E. lanceolatus ♂) was cloned and characterized, and its expression in response to dietary carbohydrate level and acute glucose treatment was investigated. The full‐length of OGT (GenBank accession no. KY656469 ) was 4,063 bp, including a 302 bp 5′untranslated terminal region (UTR), a 3,165 bp coding region that encoded 1,054 amino acids residues and a 596 bp 3′ UTR. The highly conservation of OGT gene between fish and mammals was also observed through multiple sequences alignment and phylogenetic analysis. O‐GlcNAc transferase gene was ubiquitously expressed in all detected tissues with highest expressions in brain and liver, to a lesser degree, in eye, heart, kidney and intestine. The increasing dietary carbohydrate from 8.02% to 16.08% had no significant effect on the mRNA expression of OGT. However, the expression of OGT was slightly elevated at 6 hr post‐glucose injection, and the elevation became significant at 24 hr time‐point. These data may enhance our understanding on the nutritional regulation of OGT and O‐GlcNAc modification in fish species.  相似文献   

18.
Haemocyanin is a multi‐subunit protein complex found in the haemolymph and is involved in the immune system of crustaceans. In this study, a haemocyanin gene of Macrobrachium rosenbergii, designated MrHc, was successfully isolated. The MrHc gene contained an open reading frame (ORF) of 1,992 nucleotides, encoding a protein of 663 amino acid residues with a molecular mass of 76.5 kDa. The deduced amino acid sequence contained distinct structural motifs of the haemocyanin superfamily, including an all‐alpha domain, a copper‐containing domain and an immunoglobulin‐like domain. Based on the phylogenetic analysis, the MrHC protein demonstrated a close relationship with the haemocyanins of Palaemon carinicauda and Macrobrachium nipponense. The MrHc gene was expressed in various shrimp tissues, including the hepatopancreas, gill, haemocytes, stomach and muscle. After Macrobrachium rosenbergii nodavirus (MrNV) challenge tests, the MrHc gene was up‐regulated 237‐fold at day 2. A recombinant protein of the MrHc immunoglobulin‐like domain exhibited antibacterial activity against Vibrio vulnificus, V. parahaemolyticus, Aeromonas caviae, A. veronii, A. hydrophila and Bacillus cereus. This study suggested that MrHc may play important roles in the shrimp innate immune response to MrNV infection and bacterial infection.  相似文献   

19.
Elucidation, through molecular analyses, of bacterial afflictions in commercially important aquaculture‐reared shrimps is pivotal for the prevention and/or control of disease outbreaks. In this study, we examined the phylogenetic relatedness and compared the possible immune‐related functional roles of both translationally controlled tumour protein (TCTP) and ferritin genes with previous studies. Both TCTP and ferritin genes were substantially upregulated in the Indian white prawn, Fenneropenaeus indicus (H. Milne Edwards), post‐larvae following bath challenge with the virulent strain of bacteria, Vibrio harveyi D3. Full‐length cloning of these genes by rapid amplification of complementary DNA ends ‐polymerase chain reaction (RACE‐PCR) yielded 727‐base pair (bp)‐long TCTP and 1212‐bp‐long ferritin gene sequences. Their open reading frames (ORFs) were 507 and 510 bp, respectively. The TCTP‐ORF coded for 168 amino acids with three substitutions at positions 37, 141, 155, and the ferritin ORF coded for 170 amino acids with no species‐specific substitutions. Phylogenetic analysis suggested the closest relatedness of both TCTP and ferritin from F. indicus to Chinese white prawn, Fenneropenaeus chinensis (Osbeck). In addition to reporting the full‐length sequences of these immune‐relevant genes, this study highlighted their conserved natures, which perhaps make them important defence‐related proteins in the innate immune system of F. indicus.  相似文献   

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