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Fish are extremely vulnerable to environmental stimulation and produce oxidative stress. Among them, hydrogen peroxide is an oxidative stress source that cannot be ignored in fish, which can cause physical disorders, inflammation and even death. Taurine was revealed to reduce oxidative damage and inflammation caused by toxic substances, but whether it can reduce toxicity of rice field eel caused by H 2 O 2 has not been determined. Thus, the intervention effects of taurine on H 2 O 2 -induced oxidative stress, inflammation, apoptosis, and autophagy in rice field eel. The results showed that oxidative injury in the liver was determined after H 2 O 2 injection, as indicated by enhanced serum AST and ALT activities, inhibited the antioxidant function (increased MDA and ROS contents, decreased antioxidant enzymes, inhibited nrf2 transcription level), and induced inflammatory response (upregulated il-1β, il-6, il-8, and il-12β gene expression, downregulated tgf-β1 gene expression, activated the transcription level of nf-κb, tlr-3, and tlr-7). In addition, bax, caspase3, beclin1, and Lc3B gene expression were significantly upregulated after H 2 O 2 injection, while bcl2 and p62 gene expression were downregulated, leading to the occurrence of apoptosis and autophagy. In contrast, adding 0.2 and 0.5% taurine to feed significantly alleviated this damage, as indicated by the recovery of the aforementioned bioindicators, and the effect of 0.5% taurine addition is better than 0.2%. Overall, these results suggested that taurine can relieve the liver toxicity induced by H 2 O 2 , which enriched the toxic mechanism of H 2 O 2 on fish and provided evidence for the protective effect of taurine on liver., Competing Interests: Declaration of competing interest The authors declare that they have no conflict of interest., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

Interleukin-1 receptor-associated kinase 4 (IRAK4) plays a crucial role in the Toll-like receptor/IL-1R signal pathway, which mediates the downstream signal transduction involved in innate and adaptive immunity. In the present study, an IRAK4 homologue (named as MaIRAK4) from blunt snout bream (Megalobrama amblycephala) was cloned and characterized. The open reading frame (ORF) of MaIRAK4 contains 1422 nucleotides, encoding a putative protein of 473 amino acids. Protein structural analysis revealed that MaIRAK4 has an N-terminal death domain (DD) and a central kinase domain (S_TKc), similar to those of mammals and other fishes. Multiple sequence alignment demonstrated that MaIRAK4 is highly homologous with that of grass carp (97.67%). The qRT-PCR analysis showed that MaIRAK4 expressed widely in all examined tissues, including heart, liver, spleen, kidney, head-kidney, gill, intestine and muscle, with the highest expression in the liver and spleen. After stimulation with LPS, MaIRAK4 expression upregulated significantly and reached a peak at 6 h and 12 h post LPS stimulation in the spleen and head-kidney, respectively. After challenge with Aeromonas hydrophila, MaIRAK4 expression peaked at 48 h and 72 h in spleen/head-kidney and liver, respectively. These results implied that MaIRAK4 is involved in the host defense against bacterial infection. Subcellular localization analysis indicated that MaIRAK4 distributed in the cytoplasm. Co-immunoprecipitation and subcellular co-localization assay revealed that MaIRAK4 can combine with MaMyD88 through DD domain. MaIRAK4 overexpression can induce slightly the NF-κB promoter activity in HEK 293 cells. However, the activity of NF-κB promoter was dramatically enhanced after co-transfection with MaIRAK4 and MaMyD88 plasmids. The results showed that MaIRAK4 was involved in NF-κB signal pathway mediated by maMyD88. The expression level of pro-inflammatory cytokines (IL-1β, IL-6, IL-8 and TNF-α) decreased significantly after the siRNA-mediated knockdown of MaIRAK4. Together, these results suggest that MaIRAK4 plays an important function in the innate immunity of M. amblycephala by inducing cytokines expression., (Copyright © 2022 Elsevier Ltd. All rights reserved.)

Lycium barbarum polysaccharide (LBP) is one of the main active ingredients in the fruit of L. barbarum L. It has been used as herbal medicine for thousands of years in China. In this study, Nile tilapia (Oreochromis niloticus) was taken as the research object. After feeding tilapia with 5 different doses of LBP (0 mg/kg, 500 mg/kg, 1000 mg/kg, 1500 mg/kg, 2000 mg/kg) for 55 d, it was found that LBP could promote the growth of tilapia, and this effect was the strongest at Group 1500 mg/kg. Apoptosis analysis in the liver and spleen showed that dietary supplementation with 1000 mg/kg LBP had the best protective effect on the spleen and liver in tilapia. Combined transcriptomics and metabolomics of the spleen in tilapia at Group 0 mg/kg and 1000 mg/kg showed that the differentially expressed genes (DEGs) such as NT5C2L1, pmm1, FasL and the differentially metabolites such as xanthine, dGMP, guanine and glutamate were mainly concentrated in signaling pathways such as Purine metabolism and FoxO signaling pathway. In conclusion, LBP regulates the metabolic waste levels of tilapia mainly through Purine metabolism and the FoxO signaling pathway, thereby inhibiting cell apoptosis, improving the utilization of nutrients, and promoting the growth of tilapia. This study not only provides a theoretical basis for the application of LBP in aquatic animals but also provides useful information for the healthy development of the aquaculture., (Copyright © 2021. Published by Elsevier Inc.)

Hypophthalmichthys molitrix (silver carp) is phytoplanktivorous and is an economically and ecologically important fish species. As a well-known invasive species, a number of factors associated with the ecological adaptations of this species are largely unknown. Here, we present a chromosomal-level assembly of the species based on the PacBio Sequel II platform and Hi-C scaffolding technology. Based on the high-quality genome sequences and previous genome sequencing projects, a number of genes that were probably subject to positive selection reside in the genome of H. molitrix, and the last common ancestors of H. molitrix and H. nobilis were identified. Some of these genes may partially explain the mechanisms of H. molitrix for surviving damage due to toxic algae. Demographic history estimation suggests that the effective population size (EPS) of the species may have constantly increased along with the uplift of the Qinghai-Tibet Plateau, started to decline when quaternary glaciation started, and further declined during the Younger Dryas Period. Moreover, the introgression from H. nobilis to H. molitrix in North America was corroborated based on the whole-genome sequencing data, and the proportion of introgressed regions was estimated to be approximately 5.8%. Based on the high-quality assembly, the possible mechanisms by which H. molitrix adapts to its endemic and invaded locations were profiled., (Copyright © 2021. Published by Elsevier Inc.)

DGAT2 (acyl CoA:diacylglycerol acyltransferase 2) is a key and rate-limiting enzyme that catalyzes the final step of triglyceride (TG) synthesis. In this study, hybrid tilapia were generated from Nile tilapia (♀) and blue tilapia (♂) crossing. The TG content levels in the liver of these tilapia were measured. The results showed that the TG content was higher in the hybrid tilapia. In addition, protein and mRNA expression levels in the tilapia livers were determined. Higher hepatic mRNA and protein expression of DGAT2 in the hybrid fish was found. A luciferase reporter assay with HEK293T cells revealed that miRNA-19a-5p targeted the 3'UTR of DGAT2, suggesting a direct regulatory mechanism. Using qRT-PCR, we found that DGAT2 mRNA levels had a negative correlation with miRNA-19a-5p expression in Nile tilapia and hybrid. Taken together, these findings provide evidence that miRNA-19a-5p is involved in TG synthesis in the regulation of lipid metabolism in tilapia., (Copyright © 2021 Elsevier Inc. All rights reserved.)

In this study, RNA sequencing was used to identify the hepatic gene expression profile in grass carp associated with luteinizing hormone-releasing hormone agonist (LHRH-A) treatment. A total of 93,912,172 reads were generated by HiSeq 4000 sequencing platform. After filtering, 83,450,860 clean reads were mapped to the reference genome. By calculating the FPKM of genes, 1475 differentially expressed genes were identified. PPAR signaling pathway was enriched with upregulated genes in LHRH-A injection group showing the regulation of the lipid metabolism by LHRH-A. The expression of eight key genes in PPAR signaling pathway was confirmed by qPCR and the results suggested that ACSL4A, ACSL4B, ANGPTL4, LPL, RXRBA and SLC27A1B were significantly stimulated by LHRH-A injection. This investigation provides the evidence that LHRH-A could play a role in lipid metabolism., (Copyright © 2020 Elsevier Inc. All rights reserved.)

Tumor necrosis factor receptor-associated factor 3 (TRAF3) is a multifunctional adaptor protein primarily involved in both bacterial defense and antiviral immunity in living organisms. However, the knowledge on TRAF3 in blunt snout bream (Megalobrama amblycephala), a freshwater fish with economic values, remained unclear. In the present study, we identified and characterized successfully Traf3 gene from M. amblycephala (maTraf3). The maTraf3 cDNA contained a 1722 bp open reading frame that encoded a protein of 573 amino acid residues. The deduced amino acid sequence comprised of a RING finger domain, two zinc finger motifs, a coiled-coil region and a MATH domain. Analysis of the transcriptional patterns of maTraf3 revealed that it was ubiquitously distributed in various tissues tested from M. amblycephala, with the abundance of expression in spleen and muscle. Following a challenge with Aeromonas hydrophila and lipopolysaccharide stimulation, the expression of maTraf3 was strongly enhanced at different time points in vitro and in vivo. MaTRAF3 was identified as a cytosolic protein and suggested to form aggregates or be associated with vesicles scattering in the cytoplasm. NF-κB transcription was activated by maTraf3 in reporter assay. The overexpression of maTraf3 produced high levels of pro-inflammatory cytokines such as IL-1β, IL-6, IL-8 and TNF-α, implying its immune-regulatory role in M. amblycephala. Taken together, our results obtained in this study demonstrated the crucial role of maTraf3 in mediating host innate immune response to pathogen invasion via NF-κB signaling pathway, which might indicate a novel therapeutic approach to combat bacterial infection in fish., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

Streptococcus agalactiae is a common pathogen in aquatic animals, especially tilapia, that hinders aquaculture development and leads to serious economic losses. Previously, a S. agalactiae strain named HN016 was identified from infected tilapia, and the attenuated strain YM001 was subsequently obtained by continuous passaging in Tryptic Soy Broth (TSB) medium. YM001 has been demonstrated as a safe vaccine for S. agalactiae infection in tilapia. To understand the molecular bases of the virulence of these two strains, we performed proteomic and transcriptomic analysis to reveal the protein and gene expression changes in the liver and intestine during the infection process. HN016 significantly decreased the contents of white blood cells (WBCs), neutrophils (NEUs), red blood cells (RBCs) and hematocrit (HCT) and increased the levels of total protein (TP), albumin (ALB) and globulin (GLO), while no such significant differences were observed when comparing the control with YM001. During the infection process, pathogenic peptidoglycan hydrolase, CSPA and membrane proteins were significantly differentially expressed between YM001 and HN016. Furthermore, both proteome and transcriptome data showed that the complement and coagulation cascades pathway and the antigen processing and presentation pathway were stimulated in the liver and intestine, respectively, by YM001 infection compared to HN016 infection. The interaction network analysis of key virulence genes from pathogens suggested that CSPA, as a key node, affects the expression of DOLPP1, MIPEP, PA2G4, OCIAD1, G3BP1 and CLIC5 with a positive correlation. The present evidence suggests that during the infection process, CSPA was the key genes contributing to low virulence in YM001., Competing Interests: Declaration of competing interest The authors declare no conflict of interest., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

Ghrelin is a peptide hormone secreted by gastrointestinal tract which regulates multiple physiological processes such as appetite, metabolism, growth and gonad development in fish. In the present study, the effects of ghrelin on hybrid tilapia infected with Aeromonas hydrophila are elucidated. Juvenile hybrid tilapia fish (20.0 ± 5.0 g) were intraperitoneally injected with 0, 0.1, 1.0, or 10.0 ng/g ghrelin/body weight synthetic ghrelin alone or in combination with A. hydrophila (0.5 × 10 6  CFU). At 10 days post treatment, the survival rate in the group that received 1.0 ng/g ghrelin/body weight ghrelin in combination with A. hydrophila was higher (66.66%) than that of the Ah group (13.33%) that received A. hydrophila alone. In tilapia that received ghrelin injections, reactive oxygen species (ROS) levels tended to increase at 5 h, while injection of 10.0 ng/g ghrelin/body weight ghrelin resulted in a significant decrease in ROS levels at 10 h. No changes in serum immune or antioxidant-related indicators were observed in fish injected with A. hydrophila compared to controls. However, ghrelin injection decreased Albumin (ALB), glutathione peroxidase (GSH-Px), lysozyme (LZM) and superoxide dismutase (SOD). Histological analysis showed that ghrelin injection alleviated the pathological changes in liver and spleen caused by A. hydrophila infection. Overall, the expression of HSP70, IL-1β, and TGF-β in the liver tended to upregulate compared to the control. In the kidney, HSP70, IL-1β and TGF-β levels were increased, and TNF-α expression levels were decreased compared to the control. The HSP70 level in the spleen was decreased, and IL-1β, TGF-β, and TNF-α were expressed at significantly higher levels in the spleen in the tilapia that received ghrelin injections. Taken together, our results indicate that injection with 1.0 ng/g ghrelin/body weight ghrelin may effectively protect juvenile hybrid tilapia against A. hydrophila infection by improving hematological indicators, maintaining normal histology and regulating cytokine gene expression., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

In this study, the effect of Lycium barbarum polysaccharides (LBP) on immunological parameters, apoptosis, and growth performance of Nile tilapia (Oreochromis niloticus) was investigated. Dietary supplementation with LBP significantly increased complement 3 (C3) activity and promoted interleukin IL-1β gene expression in spleen tissue, significantly reduced apoptosis in spleen tissue, increased the specific growth rate (SGR), relative length gain (LG), and relative weight gain (WG) of Nile tilapia. However, dietary supplementation with LBP did not have a significant effect on serum alkaline phosphatase (AKP), malondialdehyde (MDA), and superoxide dismutase (SOD), blood constituents, apoptosis, or gene expression of IL-1β in liver tissue. Overall, the results showed that dietary supplementation with LBP increased the nonspecific immunity of Nile tilapia and reduced the apoptosis rate to promote growth and development. Thus, LBP has potential for use as a new immunostimulant in aquaculture., Competing Interests: Declaration of competing interest We declare that we have no conflict of interest., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

Selenium is an essential element but toxic at high levels in animals. The effects of Se on growth performance and the immune system in Nile tilapia remain inconclusive. In this study, Nile tilapia Oreochromis niloticus was fed on selenium yeast (Se(Y))- and selenite (Se(IV))-enriched feed at 0, 3, 6, and 12 μg/g (dry wt) for 45 and 90 d. The growth, bioaccumulation, biochemical markers related to antioxidant, immunological, nervous and digestive systems were evaluated in various fish tissues (liver, intestine, kidney, muscle, brain, spleen, gills). The results showed that the accumulation of Se(Y) was 1.3-2 folds of Se(IV) in most tissues. The growth of tilapia was enhanced by both Se(Y) and Se(IV) at 3 μg/g after 90 d, with Se(Y) better than Se(IV) in tilapia feed. After 45 d, the levels of lipid peroxidation, the activity of the antioxidant enzymes, and the transcriptional levels of the immune related genes (IL-1β, IFN-γ and TNF-α) and stress proteins (HSP70 and MT) were enhanced in all treatments, except that of MT in the 12 μg/g Se(Y) group. In addition, both Se species inhibited the activity of acetylcholinesterase (AChE) in the brain and one digestive enzyme α-glucosidase (α-Glu) in the intestine at 12 μg/g. However, after 90 d, the effects on most biochemical markers were less pronounced, implying a possible acclimation after prolonged duration. The results demonstrate Se is beneficial to O. niloticus at low levels and toxic at elevated levels. The immunostimulation by Se might be greatly weakened after long term feeding Se-enriched feed. This study helps to better understand the effects of Se on the antioxidant and immune systems and to establish the optimal Se levels in the feed and duration for O. niloticus., Competing Interests: Declaration of competing interest The authors declare that there are no conflicts of interest in the present experiment., (Copyright © 2019 Elsevier Ltd. All rights reserved.)

Common carp (Cyprinus carpio) is a widespread freshwater fish and economically important species in China and other East Asian countries. Recent studies suggest that insulin can alter the expression of immune genes and, thus, can be regarded as an immunomodulatory hormone. To understand the mechanism of the immune response to insulin, we performed a comparative RNA-seq transcriptome analysis using livers from common carp injected with insulin (5 μg/g bodyweight) or saline as a control. After filtering the low-quality reads and removing the adaptors, the clean raw reads were assembled into 60,421 unigenes with mean length of 746.81 bp. Furthermore, 37,107 unigenes were annotated based on homology after blast search in public databases. Differentially expressed genes were identified using the fragments per kb per million fragments method and EdgeR software. In total, 782 differentially expressed genes were found. Thereinto, 444 and 338 genes were upregulated and downregulated, respectively, in the insulin-injected group. A Gene Ontology analysis indicated that these genes were concentrated in glucose metabolism, hormone secretion, andimmune system processes. Moreover, 153 enriched KEGG pathways were associated with the differentially expressed genes, including the Toll-like receptor (TLR) and nuclear factor kappa beta (NF-κB) signaling pathways. Signal transducer and activator of transcription 1 (10.56-fold), TLR3 (0.089-fold), activator protein-1 (0.007-fold), tumor necrosis factor-α (0.139-fold), and macrophage inflammatory protein-1β (0.038-fold) expression were significantly changed after the insulin injection. This study characterized the profile of genes expression response to insulin in common carp liver for the first time and provided new insight into understanding the molecular mechanism of insulin as an immunomodulatory hormone., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

Toll-like receptors (TLRs) are central players in the innate immune system in response to a wide range of pathogen infection. Among various TLRs, TLR4 plays a key role in recognition of bacterial lipopolysaccharides (LPS). In the present study, we identified and characterized a novel TLR4 homologue (maTLR4b) in blunt snout bream (Megalobrama amblycephala) which was significantly distinct from previously reported M. amblycephala TLR4 (tentatively named maTLR4a). The results showed that the complete cDNA sequence of maTLR4b was 3261 bp with an open reading frame encoding a polypeptide of 820 amino acids, and that its genomic sequence was 3793 bp, which had 3 exons. Structurally, the deduced maTLR4b protein showed a typical TLR domain architecture, including a signal peptide, eight leucine-rich repeats (LRRs) in the extracellular region, a transmembrane domain, and a Toll-Interleukin 1 receptor (TIR) domain in the cytoplasmic region. Phylogenetic analysis revealed that all TLR4s from teleost fish formed a monophyletic clade. Both maTLR4a and maTLR4b were divided into two distinct branches, and showed the highest level of similarity with the grass carp TLR4.2 and TLR4.4 homologue, respectively. MaTLR4b was constitutively expressed in all healthy tissues tested although at different levels. After LPS stimulation, the expression levels were significantly up-regulated in spleen, and peaked at 4 h between maTLR4a and maTLR4b, but with a distinct and complementary expression patterns. Taken together, these results suggested that maTLR4b is indeed a functional homologue of TLR4 in other species, which may play vital role in innate immune., (Copyright © 2016 Elsevier Ltd. All rights reserved.)

Bactericidal permeability-increasing protein (BPI) and lipopolysaccharide-binding protein (LBP) belong to the lipid transfer protein/lipopolysaccharide-binding protein family and play a critical role in the innate immune response to Gram-negative bacteria. In the present study, a novel BPI/LBP from blunt snout bream, Megalobrama amblycephala (maBPI/LBP) was isolated by RACE techniques. The open reading frame (ORF) of maBPI/LBP gene encoded a polypeptide of 474 amino acids with a putative 18-aa hydrophobic signal peptide. Structurally, the maBPI/LBP showed highly similar to those of BPI/LBPs from invertebrate and teleost, LBPs and BPIs from mammal, which contained an N-terminal BPI/LBP/CETP domain BPI1 with a LPS-binding domain, a C-terminal BPI/LBP/CETP domain BPI2, and proline-rich domain. The homologous identities of deduced amino acid sequences displayed that the maBPI/LBP possessed significant similarity (96.61% and 90.07%) with those of grass carp and common carp, respectively. The recombinant protein of maBPI/LBP showed effectively kill Gram-negative bacteria. The maBPI/LBP gene was expressed in a wide range of normal tested tissues, with the highest expression levels in the kidney. The experiments revealed that the mRNA expression of maBPI/LBP in spleen considerably up-regulated from 2 h to 8 h post LPS stimulation, and peaked rapidly at 2 h (7.40-fold, P < 0.05), which confirmed that maBPI/LBP was the absolute sensitive to LPS stimulation. Furthermore, the level of maBPI/LBP mRNA expression reached the maximum for a second time at 24 h after LPS stimulation. These results suggested that maBPI/LBP was a constitutive and inducible acute-phase protein contributing to the host immune defense against pathogenic bacterial infection in M. amblycephala. This study will further our understanding of the function of BPI/LBP and the molecular mechanism of innate immunity in teleost., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

High-throughput sequencing of transcriptome promises a new approach for detecting evolutionary divergence among species. Up to now, the information about evolution of immune genes in cultured fish, especially in tilapias which would aid to understand the molecular basis of immune phenotypic differentiation is still lack. Thus, in the present study, we used high-throughput sequencing to obtain large amount of gene sequences in blue tilapia and characterized the diversity of orthologs among Nile tilapia, blue tilapia and zebrafish. A total of 52,424,506 raw reads, representing 31,404 unigenes were obtained from blue tilapia cDNA library of mixed tissues, including brain, pituitary, gill, heart, liver, spleen, kidney, intestine, muscle, testis and ovary. Based on Ks value, we calculated that the divergence time between Nile tilapia and blue tilapia is 2.93 million years ago. And the tilapias are both apart from zebrafish in 197 million years ago. Furthermore, the positive selected genes were identified by calculating of Ka/Ks ratio. Several immune-related genes were identified as positively selected genes, such as Notch2 and nfatc3b. Considering that these genes play crucial role in immune regulating function, the immune system genes met a great variation under environment selection in tilapias which suggests fast evolution in immune system of cultured tilapias., (Copyright © 2015 Elsevier Ltd. All rights reserved.)

We profiled the expression of a group of proinflammatory immune genes, comprising TNFα-1, TNFα-2, IFN-γ, IL1β-1, IL1β-2, CCL-1, and CXCL-8 in liver, head kidney, gills, and spleen of goldfish, during the reproductive cycle and in response to injection of the hormone human chorionic gonadotrophin (hCG). Most genes showed higher expression during the breeding season in both sexes. However, activation of immune responses was much stronger in female goldfish. Injection with hCG, an analog of luteinizing hormone (LH), which is involved in numerous reproductive functions, markedly changed gene expression in most studied organs, in both male and female goldfish. Again, female goldfish were found to be more responsive than male goldfish. The strongest activation of these genes was seen 7 days post-injection; the effect was dose dependent with a lower dose being in general more effective. For several of the genes, the gills were the most responsive tissue and, in male goldfish, gills were often the only responsive tissue, suggesting an important immunological role for gills during breeding. The data suggest that increasing expression levels are regulated by LH arising during the breeding season, with greater sensitivity in female goldfish than in male goldfish. These data support an interaction between the innate immune system and the reproductive axis., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

Piwi proteins as constituent factors of the piRNA pathway are required for germline maintenance, meiosis and gonad development. Previous study showed hCG could regulate the Piwi expression in ovary of teleosts. In this study, we revealed effects of LHRH-A and hCG on Piwi expression in testis of tilapia using Real-time PCR and Western blot. Both in vivo and in vitro study suggest that LHRH-A and hCG significantly down-regulated Piwil-1 and Piwil-2 in mRNA or protein levels compared with controls. Meanwhile, tissue and cell distribution showed that Piwi proteins were mainly expressed in spermatocytes rather than mature sperms. These results indicated that HPG suppresses Piwis which may play a crucial role in testis differentiation and development., (Copyright © 2013 Elsevier Inc. All rights reserved.)

Growth hormone (GH), growth hormone receptor (GHR) and insulin-like growth factor 1 (IGF-1) are pivotal signaling factors of the GH/IGF axis, which plays a crucial role in regulating growth in vertebrates. In this study, GH, GHR and IGF-1 cDNAs were cloned from triploid and tetraploid crucian carp. In addition, mRNA expression levels were characterized in diploid red crucian carp, triploids and tetraploids. Reverse transcriptase PCR indicated that GH genes were only expressed in the pituitary, while GHR and IGF-1 were widely expressed in all tested tissues. Real-time PCR study of different seasonal profiles showed that triploids had significantly higher expression of the studied genes during both the prespawning and the spawning season. Although different temperatures (22, 26 and 30°C) showed no significant effects on GH, GHR and IGF-1 mRNA expression in either diploids or triploids, triploids had higher expression levels than diploids at each temperature. After 1 week of fasting, the expression of all studied genes was reduced in both diploids and triploids, while the expressions levels were higher in triploids than in diploids. These results suggest that the elevated expression of GH/IGF axis genes in triploids plays a crucial role in the faster growth rate of triploids., (Copyright © 2012 Elsevier Inc. All rights reserved.)

Piwi proteins are required for germline maintenance and gonad development. In this study, the cDNAs encoding Piwil1 and Piwil2 were cloned and sequenced from the common carp. The full-length cDNA of Piwil1 and Piwil2 were 3114 and 3421bp, encoding 858 and 1034 amino acids including PAZ domain and PIWI domain, respectively. In addition, the Piwil1 and Piwil2 proteins shared high homology with other teleosts. Reverse transcriptase PCR revealed that the Piwi mRNAs were exclusively expressed in adult testes and ovaries. Using real-time PCR, expression study of different developmental profiles showed that Piwil1 and Piwil2 were down-regulated during pre-ovulation. Further, human chorionic gonadotropin treatment in ovaries (in vivo) and in cultured ovaries cells (in vitro) resulted in down-regulation of Piwi RNAs. These results suggest that the decreased expression which was regulated by hormone plays a crucial role during ovarian differentiation and development., (Copyright © 2012 Elsevier Inc. All rights reserved.)