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Natural and synthetic environmental estrogens (EEs) are widespread and have received extensive attention. Our previous studies demonstrated that depletion of the cytochrome P450 17a1 gene (cyp17a1) leads to all‐testis differentiation phenotype in zebrafish and common carp. In the present study, cyp17a1‐deficient zebrafish with defective estrogen biosynthesis were used for the evaluation of EEs, as assessed by monitoring vitellogenin (vtg) expression. A rapid and sensitive assessment procedure was established with the 3‐day administration of estradiol (E2), followed by examination of the transcriptional expression of vtgs in our cyp17a1‐deficient fish. Compared with the control fish, a higher E2‐mediated vtg upregulation observed in cyp17a1‐deficient zebrafish exposed to 0.1 μg/L E2 is known to be estrogen receptor–dependent and likely due to impaired in vivo estrogen biosynthesis. The more responsive vtg expression in cyp17a1‐deficient zebrafish was observed when exposed to 200 and 2000 μg/L bisphenol A (BPA) and perfluoro‐1‐octanesulfonate (PFOS). The estrogenic potentials of E2, BPA, and PFOS were compared and assessed by the feminization effect on ovarian differentiation in cyp17a1‐deficient zebrafish from 18 to 50 days postfertilization, based on which a higher sensitivity of E2 in ovarian differentiation than BPA and PFOS was concluded. Collectively, through the higher sensitivity to EEs and the capacity to distinguish chemicals with different estrogenic potentials exhibited by the all‐male cyp17a1‐deficient zebrafish with impaired estrogen biosynthesis, we demonstrated that they can be used as an excellent in vivo model for the evaluation of EEs. Environ Toxicol Chem 2024;43:1062–1074. © 2024 SETAC [ABSTRACT FROM AUTHOR]

(1) Background: Non-small cell lung cancer (NSCLC) is the most common lung cancer. Enhancer RNA (eRNA) has potential utility in the diagnosis, prognosis and treatment of cancer, but the role of eRNAs in NSCLC metastasis is not clear; (2) Methods: Differentially expressed transcription factors (DETFs), enhancer RNAs (DEEs), and target genes (DETGs) between primary NSCLC and metastatic NSCLC were identified. Prognostic DEEs (PDEEs) were screened by Cox regression analyses and a predicting model for metastatic NSCLC was constructed. We identified DEE interactions with DETFs, DETGs, reverse phase protein arrays (RPPA) protein chips, immunocytes, and pathways to construct a regulation network using Pearson correlation. Finally, the mechanisms and clinical significance were explained using multi-dimensional validation unambiguously; (3) Results: A total of 255 DEEs were identified, and 24 PDEEs were selected into the multivariate Cox regression model (AUC = 0.699). Additionally, the NSCLC metastasis-specific regulation network was constructed, and six key PDEEs were defined (ANXA8L1, CASTOR2, CYP4B1, GTF2H2C, PSMF1 and TNS4); (4) Conclusions: This study focused on the exploration of the prognostic value of eRNAs in the metastasis of NSCLC. Finally, six eRNAs were identified as potential markers for the prediction of metastasis of NSCLC. [ABSTRACT FROM AUTHOR]

In this systematic review, we highlight the differences between the male and female zebrafish brains to understand their differentiation and their use in studying sex-specific neurological diseases. Male and female brains display subtle differences at the cellular level which may be important in driving sex-specific signaling. Sex differences in the brain have been observed in humans as well as in non-human species. However, the molecular mechanisms of brain sex differentiation remain unclear. The classical model of brain sex differentiation suggests that the steroid hormones derived from the gonads are the primary determinants in establishing male and female neural networks. Recent studies indicate that the developing brain shows sex-specific differences in gene expression prior to gonadal hormone action. Hence, genetic differences may also be responsible for differentiating the brain into male and female types. Understanding the signaling mechanisms involved in brain sex differentiation could help further elucidate the sex-specific incidences of certain neurological diseases. The zebrafish model could be appropriate for enhancing our understanding of brain sex differentiation and the signaling involved in neurological diseases. Zebrafish brains show sex-specific differences at the hormonal level, and recent advances in RNA sequencing have highlighted critical sex-specific differences at the transcript level. The differences are also evident at the cellular and metabolite levels, which could be important in organizing sex-specific neuronal signaling. Furthermore, in addition to having one ortholog for 70% of the human gene, zebrafish also shares brain structural similarities with other higher eukaryotes, including mammals. Hence, deciphering brain sex differentiation in zebrafish will help further enhance the diagnostic and pharmacological intervention of neurological diseases. [ABSTRACT FROM AUTHOR]

Hierarchically porous carbon sheets decorated with transition metal carbides nanoparticles and metal-nitrogen coordinative sites have been proposed as the promising non-precious metal oxygen electrocatalysts. In this work, we demonstrate a facile and low-cost strategy to in situ form Fe/N codoped hierarchically porous graphene-like carbon nanosheets abundant in Fe-N x sites and Fe 3 C nanoparticles (Fe–N/C) from pyrolyzing chestnut shell precursor. The as-prepared Fe–N/C samples with abundant Fe-N x sites and Fe 3 C nanoparticles show superior electrocatalytic activity to oxygen reduction reaction (ORR) in the alkaline medium as well as high stability and methanol tolerance due to the integration of multi-factors: the high content of Fe-N x active sites, the coexistence of Fe 3 C, the unique hierarchically porous structure and high conductivity of carbon matrix. The optimal Fe–N/C-2-900 sample exhibits a more positive half-wave potential (−0.122 V vs. Ag/AgCl (3 M) reference electrode) than commercial 20 wt% Pt/C catalyst. This study provides a facile approach to synthesize Fe 3 C nanoparticles decorated Fe/N co-doped hierarchically porous carbon materials for effective oxygen electrocatalyst. • Hierarchically porous graphene-like carbon nanosheets were successfully synthesized. • The highly graphitized carbon nanosheets are abundant in Fe related active sites (Fe-N x and Fe 3 C). • The optimal sample can act as an effective oxygen electrocatalyst for ORR. [ABSTRACT FROM AUTHOR]

Daphnia sinensis is a widespread freshwater microcrustacean. The assembled D. sinensis genome totaled 131.58 Mb with 92.23% of the assembly anchored onto 10 chromosomes. Based on the whole genome information, we further compared the transcriptomic and epigenomic characterization among parthenogenetic females, sexual females and males in D. sinensis. Transcriptomic analysis showed that the up-regulated genes in males were mainly grouped into the cuticle, sex differentiation and methyl farnesoate synthesis, which might play a pivotal role in steering development and reproduction processes. By comparison, the highly expressed genes in parthenogenetic females were mainly grouped into energy metabolism, mitosis, and DNA replication, which might contribute to maintaining rapid production of parthenogenetic females, and nutrient uptake for the growth of neonates. The whole-genome DNA methylation analysis showed that the methylation rate in parthenogenetic females was higher than that in sexual females and males, which might contribute to its rapid response to environment stress. • D. sinensis genome was 131.58 Mb with 92.2% of assembly anchored onto 10 chromosomes. • High temperature and population density could induce reproductive switching. • D. sinensis could revert to a parthenogenetic mode after producing resting eggs. • Up-regulated genes in males were grouped into sex differentiation and MF synthesis. • High methylation rate in PF contributed to its rapid response to environment stress. [ABSTRACT FROM AUTHOR]

d-Allulose is the corresponding epimer of d-fructose at the C-3 position, which exhibits a similar taste and sweetness to sucrose. As a low-calorie sweetener, d-allulose has broad application prospects in the fields of medicine, food, and so on. Currently, the production method of d-allulose is mainly the enzymatic conversion of d-fructose by d-allulose 3-epimerase (DAEase). However, the limited specific activity and thermal stability of DAEase restrict its industrial application. Herein, an ultrahigh-throughput screening assay based on the transcription factor PsiR was extensively optimized from the aspects of culture medium components, screening plasmid, and expression host, which enhanced the correction between the fluorescent readout and the enzyme activity. Then, the error-prone PCR (epPCR) library of Clostridium cellulolyticum H10 DAEase (CcDAEase) was screened through the above optimized method, and the variant I228V with improved specific activity and thermal stability was obtained. Moreover, after combining two beneficial substitutions, D281G and C289R, which were previously obtained by this optimized assay, the specific activity of the triple-mutation variant I228V/D281G/C289R reached up to 1.42-fold of the wild type (WT), while its half-life (T1/2) at 60 °C was prolonged by 62.97-fold. The results confirmed the feasibility of the optimized screening assay as a powerful tool for the directed evolution of DAEase. [ABSTRACT FROM AUTHOR]

There are several technical challenges and public issues concerning genome editing applications before they become viable in commercial aquaculture. Recently, we developed a novel strategy to generate all-female (AF) common carp, which exhibited a growth advantage over the control carp, using genetic editing through single gene-targeting manipulation. Here, we found that the body weight of the AF common carp was higher by 22.58% than that of the control common carp. Because the genotype of the AF common carp was cyp17a1 +/−;XX, the contents of sex steroids were normally synthesized, as they were comparable to that of the control female carp. To evaluate the food safety of the AF carp, Wistar rats were fed a diet containing control female carp (control, C) or all-female (AF) carp at an incorporation rate of 5, 10 and 20% (w/w) for 90 days. Compared with those fed control carp, the rats fed AF common carp exhibited no significant difference in body weight, food intake, feed conversion ratio, hematology, serum biochemistry, urine test, relative organ weight, gross necropsy, and histopathological examination. This is the first food safety assessment of the farmed fish strain cultured using CRISPR/Cas9, which will further advance the fishery development of genome-edited animals. • The body weight of AF common carp was higher by 22.58% than that of the control carp. • Feeding of AF carp to Wistar rats did not cause adverse effect in the parameters evaluated due to subchronic toxicity. • We provide the scientific evidence for the food safety assessment of the farmed fish strain cultured using CRISPR/Cas9. [ABSTRACT FROM AUTHOR]

Bioinspired scaffolds with two distinct regions resembling stratified anatomical architecture provide potential strategies for osteochondral defect repair and are studied in preclinical animals. However, delamination of the two layers often causes tissue disjunction between the regenerated cartilage and subchondral bone, leading to few commercially available clinical applications. This study develops an integrated poly(ε‐caprolactone) (PCL)‐based scaffold for repairing osteochondral defects. An extracellular matrix (ECM)‐incorporated 3D printing composite scaffold (ECM/PCL) coated with ECM hydrogel (E‐co‐E/PCL) is fabricated as the upper layer, and magnesium oxide nanoparticles coated with polydopamine (MgO@PDA)‐incorporated composite scaffold (MD/PCL) is fabricated using 3D printing as the bottom layer. The physicochemical and mechanical properties of the bilayer scaffold meet the requirements in designing and fabricating the osteochondral scaffold, especially a strong interface possessed between the two layers. By in vitro study, the integrated scaffold stimulates proliferation, chondrogenic differentiation, and osteogenic differentiation of human bone mesenchymal stem cells. Moreover, the integrated bilayer scaffold exhibits well repair ability to facilitate simultaneous regeneration of cartilage and subchondral bone after implanting into the osteochondral defect in rats. In addition, cartilage "tidemarks" completely regenerated after 12 weeks of implantation of the bilayer scaffold, which indicates no tissue disjunctions formed between the regenerated cartilage and subchondral bone. [ABSTRACT FROM AUTHOR]

Aims: Opioid addiction is a major public health issue, yet its underlying mechanism is still unknown. The aim of this study was to explore the roles of ubiquitin–proteasome system (UPS) and regulator of G protein signaling 4 (RGS4) in morphine‐induced behavioral sensitization, a well‐recognized animal model of opioid addiction. Methods: We explored the characteristics of RGS4 protein expression and polyubiquitination in the development of behavioral sensitization induced by a single morphine exposure in rats, and the effect of a selective proteasome inhibitor, lactacystin (LAC), on behavioral sensitization. Results: Polyubiquitination expression was increased in time‐dependent and dose‐related fashions during the development of behavioral sensitization, while RGS4 protein expression was not significantly changed during this phase. Stereotaxic administration of LAC into nucleus accumbens (NAc) core inhibited the establishment of behavioral sensitization. Conclusion: UPS in NAc core is positively involved in behavioral sensitization induced by a single morphine exposure in rats. Polyubiquitination was observed during the development phase of behavioral sensitization, while RGS4 protein expression was not significantly changed, indicating that other members of RGS family might be substrate proteins in UPS‐mediated behavioral sensitization. [ABSTRACT FROM AUTHOR]

Advanced hierarchical hybrid nanostructures of polar, conductive and catalytic chemical traps/carbon with rationally tailored components, structures, and chemistries are important in determining the performance of lithium-sulfur (Li–S) batteries. Herein we propose a facile strategy to prepare 3D-structured CNF@graphene-CoSe (CNF@G L -CoSe) cable/sheath heterostructure with the in-situ formed CoSe as a catalyst for graphene-like carbon growth and Se as a catalyst promoter. The architecture integrates high electrical conductivity, effective ion transport, notable polysulfide entrapment, and polysulfide redox kinetics, availing lithium/sulfur conversion when applied as an interlayer in a Li–S battery. As expected, the battery delivered a low decay of 0.053% per cycle over 700 cycles at 1 C. The capacity even remained 649.4 and 524.5 mA h g−1 at 4 and 6 C, respectively. Furthermore, an areal capacity of about 3 mA h cm−2 at 0.1 C after long-term tests can still maintain with sulfur loading of 4.5 mg cm−2. A facile effective strategy to prepare 3D-structured CNF@graphene-CoSe cable/sheath heterostructure with the in situ formed CoSe as a catalyst for graphene-like carbon growth and Se as a catalyst promoter, achieving significant enhancement in electrochemical performance of Li–S batteries. [Display omitted] • In situ constructing CNF@graphene-CoSe cable/sheath heterostructure. • CoSe catalyzes the growth of graphene sheath and Se as a catalyst promoter. • Synergism of conductivity, absorption and catalysis. • Enabling excellent performance as an interlayer. [ABSTRACT FROM AUTHOR]

• In situ self-boosting catalytic synthesis of hierarchical CNF-CNT/TMS architecture. • CNF/CNT doped with rich N, S. • TMS as the catalyst for secondary CNT branches growth. • Integrating of conductivity, absoption and catalysis. • Ultrastable long-life (1000 cycles) and high-rate performance (6C). Advanced hierarchical carbon-based hybrid (CH) nanostructures, typical carbon nanotubes/carbon nanofibers hybrid (CNT-CNF), with rationally tailored components, structures, and chemistries, have been widely dedicated to potential energy storage and conversion applications. Here we report a facile methodology for the synthesis of precise hierarchical heteroatom-doped carbon hybrid / transition metal sulfide (CNF-CNT/TMS) nanostructure assemblies with novel in-situ formed TMSs as the catalysts for secondary CNTs growth via a self-boosting catalytic pyrolysis. The key to in situ growth of CNT arrays is obtaining small high active TMS nanocatalysts and sufficient supply of carbon sources originated from polystyrene. Such special structure provides abundant active sites, beneficial for the application of N, S codoped CNF-CNT/TMS hybrid film in the electrochemistry without introducing extra dopants. As a proof-of-concept application, the representative CNF-CNT/Co 9 S 8 hybrid was exploited as a conductive and catalytic membrane reactor in a lithium–sulfur (Li-S) battery. This unique architecture can achieve a maximized synergistic effect of electron/ion transport, polysulfide entrapment and catalytic conversion, which enables significant enhancement in electrochemical performance of Li-S batteries. As anticipated, the Li-S battery exhibits ultrastable long-life (over 1000 cycles, with an ultralow decay of 0.034% per cycle) and high-rate performance (6 C). Moreover, at high loading of sulfur of 3.7 mg cm−2, stable cycling performance with high capacity retention can still be achieved after long-term tests. This work also presents a facile and effective strategy in the development of complex hierarchical carbon nanostructure, which may be a competitive candidate in other energy storage and conversion system. [Display omitted] [ABSTRACT FROM AUTHOR]

Colors are important phenotypic traits for fitness under natural conditions in vertebrates. Previous studies have reported several functional genes and genetic variations of pigmentation, but the formation mechanisms of various skin coloration remained ambiguous in fish. Jinbian carp, a common carp variant, displays two colors (yellow and black) in the skin, thus, it is a good model for investigating the genetic basis of pigmentation. In the present study, using the Jinbian carp as model, isobaric tags for relative and absolute quantification (ITRAQ) proteomics analysis was performed for yellow and black skin, respectively. The results showed that 467 differentially expressed proteins (DEPs) were identified between the yellow skin and the black skin. Similar to mammals, the up-regulated DEPs in black skin included UV excision repair protein RAD23 homolog A (Rad23a), melanoregulin (mreg), 5,6-dihydroxyindole-2-carboxylic acid oxidase5 (tyrp1) and melanocyte protein PMEL (PMEL), which were mainly grouped into melanogenesis pathway. However, several up-regulated DEPs in yellow skin were mainly enriched in nucleotide metabolism, such as GTPase IMAP family member 5 (GIMAP5), AMP deaminase 1 (AMPD1), adenosylhomocysteinase b (ahcy-b), and pyruvate kinase (PKM). In summary, several candidate proteins and their enrichment pathways for color variation in Jinbian carp were identified, which may be responsible for the formation of different colorations. [ABSTRACT FROM AUTHOR]

Myocardial infarction (MI) is one of the most serious cardiovascular diseases, characterized by a rapid and irreversible decline in myocardial function. Early detection of patients with MI and prolonging the optimal therapeutic window of acute myocardial infarction (AMI) are particularly important. This study aimed to identify the diagnostic biomarkers and novel therapeutic targets for acute myocardial infarction. We generated the AMI mouse models by ligating the proximal left anterior descending coronary artery. Six time points—Sham, AMI 10-min, 1-h, 6-h, 24-h, and 72-h—were chosen to examine the molecular changes that occur during the AMI process. RNA-seq and DIA-MS were performed on the infarcted left ventricular tissues of AMI mice at each time point. Co-expression pattern genes were screened from myocardial infarction samples at different time points by time-series analysis. Gene Ontology (GO) function and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis were used to examine these genes. Using the Interactive Gene/Protein Retrieval Tool (STRING) database, the protein-protein interaction network (PPI) was constructed and the hub genes were identified. In order to evaluate the diagnostic value of hub genes, a receiver operating characteristic (ROC) curve was constructed. An independent data set, GSE163772, confirmed the diagnostic value of hub genes further. We obtained the expression profiles at different time points after the occurrence of heart failure through high-throughput sequencing, and found 167 genes with similar expression patterns through time series analysis. The immune response and immune-related pathways had the greatest enrichment of these genes. Among them, Itgb2 Syk, Tlr4, Tlr2, Itgax, and Lcp2 may play key roles as hub genes. Combined with the results of proteomic analysis, it was found that the expression of Coro1a in both omics increased with time. The results of external validation showed that TLR2, ITGAX, and LCP2 had good predictive ability for AMI diagnosis. Itgb2, Syk, Tlr4, Tlr2, Itgax, Lcp2 and Coro1a are considered to be the seven key genes significantly associated with AMI. Our results may provide potential targets for the prevention of adverse ventricular remodeling and the treatment of AMI. • To clarify the pathological mechanism for AMI, we established AMI mice models and sequenced their RNA-seq and DIA-MS profiles. • The immunoinflammatory molecules with continuous action after myocardial infarction were obtained by time-series analysis b. • Itgb2, Syk, Tlr4, Tlr2, Itgax, Lcp2 and Coro1a were considered to be the seven key genes significantly associated with AMI. • Our results may provide potential targets for the prevention of adverse ventricular remodeling and the treatment of AMI. [ABSTRACT FROM AUTHOR]

Abstract The design of porous carbon frameworks with high-density distributions of dopant sites, especially those derived from renewable biomaterials, is highly desirable to improve the activity of nonprecious porous carbon catalysts for oxygen reduction reaction (ORR). In this paper, we prepared N, P-doped hierarchical porous carbon catalysts via a direct pyrolysis of a gelatin/phytic acid supermolecular aggregate without extra activation processes. The optimal catalyst materials (NPC1000) showed not only the highest N and P contents but also the highest specific surface area, total pore volume and mesoporous pore volume, which are also higher than those of the contrast sample prepared with a chitosan/phytic acid supermolecular aggregate. Thus, the sample NPC1000 had the best electrocatalytic activity compared with the Pt/C catalyst, which was nearly a direct four-electron reaction. The sample also had higher stability and methanol tolerance than Pt/C. This work provides a simple, economical and environmentally friendly method to prepare nonprecious porous carbon catalysts with the desired properties. Graphical abstract Image 1 Highlights • Biological materials gelatin and phytic acid are used as raw materials. • Porous carbon framework featuring sheets own large surface area and pore volume. • The one-step pyrolysis method is facile without activation. • The high ORR activity is attributed to high N and P content and unique structure. • Especially, the mesoporosity play a crucial role in boosting the ORR performance. [ABSTRACT FROM AUTHOR]

Large-sized porous glucose-derived carbon (C) and graphene (G) co-modified LiFePO 4 (LFP) composite microspheres (C@LFP/G) have been successfully in situ synthesized by one-pot solvothermal and followed annealing process using biological phytic acid (PhyA) as phosphorus source. The loose and porous microspheres with diameters in the range of 5–12 μm are self-assembled by carbon coated LFP nano particles along with randomly embedded/distributed graphene sheets. The effect of PhyA dosage and carbon modification on morphology and electrochemical properties were investigated. The results indicated that PhyA dosage plays an important role in construction of the self-assembled three-dimensional spherical microstructures during hydrothermal process. A reasonable assembly process elucidating the formation of the different structure is provided based on the experimental results. Additionally, the effects of carbon modification were also investigated, which reflected that the graphene and glucose-derived carbon personate a synergistic effect in regulating the size of LFP primary nanoparticles. The typical large-sized C@LFP/G microspheres composite exhibited the best electrochemical properties with the discharge specific capacity of 163.7 mA h g −1 at 0.1 C, excellent rate capability and cycling performance. This method may be extended to prepare other spherical olivine structured materials with large size as cathode materials for lithium-ion batteries. [ABSTRACT FROM AUTHOR]

• A culture system was developed to breed cladocerans. • Phylogenetic position of D. sinensis was determined. • Phylogenetic analysis of eight Anomopoda was carried out. • Phylogenetic analysis basing on transcriptome was available in cladoceran. Anomopoda is the widespread planktonic microcrustacean, which plays a crucial role in aquatic ecosystem. There are few studies about the evolutionary relationships among various Anomopoda basing on molecular data. In the present study, phylogenetic analysis of eight Anomopoda was carried out. Firstly, the culture system was developed to breed cladocerans. By using this system, eight species (Daphnia magna , D. pulex, D. sinensis , Ceriodaphnia reticulata , Moina micrura , Scapholeberis kingi, Simocephalus vetulus and Eurycercus lamellatus) were purified and cultured stably in the laboratory. Then, transcriptomic sequences and partial mitochondrial DNA sequences were both used to reconstruct the phylogenetic tree among 8 species. Transcriptomic sequences were sequenced on Illumina Hiseq 2500 platform. After assembly and annotation, transcriptomic sequences were spliced together and aligned for phylogenetic analysis. Basing on the orthologous genes derived from transcriptomic sequences, the phylogenetic analysis showed that 4 genera of Daphniidae were clustered into one group, and among the 4 genera, Ceriodaphnia was closer to Daphnia than Simocephalus , while Scapholeberis was farthest from other species. In addition, Eurycercidae was closer to Daphniidae than Moinidae. The phylogenetic trees based on both 12S rRNA and 16S rRNA sequences were similar with that based on transcriptomic sequences. Meanwhile, the phylogenetic tree based on 16S rRNA sequences was more suitable than that based on 12S rRNA sequences. These results suggested that the phylogenetic analysis basing on the transcriptomic sequences was available in cladocerans, which will help us to effectively understand the phylogenetic relationships among various cladocerans. [ABSTRACT FROM AUTHOR]

Neurokinin B (NKB), the gene product of tachykinin 3 (TAC3), has recently emerged as a key regulator for reproduction. However, previous studies were mainly focused on the functions of NKB on hypothalamus GnRH regulation in mammals, little information is available regarding its functional role at the pituitary level in teleost. In this study, using grass carp pituitary cells as model, transcriptome analysis revealed that the differential expression genes (DEGS) induced by NKBa were mainly involved in the regulation of appetite, hormone activity, growth factor, and metabolic process. Secondly, by using qPCR, we further confirmed that NKB could significantly induce four anorectic peptides (UTS1, CART, POMCb and NMB) mRNA expression in a time- and dose-dependent manner in grass carp pituitary cells. Thirdly, using a pharmacological approach, the stimulatory effects of NKBa on pituitary UTS1, CART, POMCb and NMB mRNA expression were found to be all mediated by NK3R and coupled to the cAMP/PKA, PLC/IP 3 /PKC and Ca2+/CaM/CaMK-II signal pathways. Furthermore, intraperitoneal injection of NKBa could also significantly induce the four anorectic peptides (UTS1, CART, POMCb and NMB) mRNA expression in grass carp pituitary. Finally, we found that food intake in grass carp could significantly induce hypothalamus TAC3a and TAC3b mRNA expression. These results, taken together, suggested that NKB should be a satiety factor and involved in the regulation of pituitary anorectic peptides in teleost. • The anorectic peptides (NMB, UTS1, POMCb, CART) were highly expressed in pituitary. • NKB could induce NMB, UTS1, POMCb, CART mRNA expression in grass carp pituitary. • Food intake could induce TAC3 (precursor of NKB) mRNA expression in grass carp. • NKB might be a coupling factor to integrate the feeding metabolism and reproduction. [ABSTRACT FROM AUTHOR]

Simple Summary: In mammals, the functions of epidermal growth factor (EGF) have been widely studied. However, little is known about the pituitary actions of EGF in teleost. Using primary cultured grass carp pituitary cells as model, we found that EGF could reduce pituitary luteinizing hormone β (LHβ) mRNA expression, but induce pituitary growth hormone (GH), prolactin (PRL) and somatolactins (SL) mRNA expression. Furthermore, we also found that NKB could suppress EGF-induced PRL mRNA expression in grass carp pituitary cells. These results suggested that EGF could directly regulate pituitary hormones expression in teleost. In mammals, epidermal growth factor (EGF) plays a vital role in both pituitary physiology and pathology. However, the functional role of EGF in the regulation of pituitary hormones has rarely reported in teleost. In our study, using primary cultured grass carp pituitary cells as an in vitro model, we examined the effects of EGF on pituitary hormone secretion and gene expression as well as the post-receptor signaling mechanisms involved. Firstly, we found that EGF significantly reduced luteinizing hormone (LHβ) mRNA expression via ErbB1 coupled to ERK1/2 pathway, but had no effect on LH release in grass carp pituitary cells. Secondly, the results showed that EGF was effective in up-regulating mRNA expression of growth hormone (GH), somatolactin α (SLα) and somatolactin β (SLβ) via ErbB1 and ErbB2 and subsequently coupled to MEK1/2/ERK1/2 and PI3K/Akt/mTOR pathways, respectively. However, EGF was not effective in GH release in pituitary cells. Thirdly, we found that EGF strongly induced pituitary prolactin (PRL) release and mRNA expression, which was mediated by ErbB1 and subsequent stimulation of MEK1/2/ERK1/2 and PI3K/Akt/mTOR pathways. Interestingly, subsequent study further found that neurokinin B (NKB) significantly suppressed EGF-induced PRL mRNA expression, which was mediated by neurokinin receptor (NK2R) and coupled to AC/cAMP/PKA signal pathway. These results suggested that EGF could differently regulate the pituitary hormones expression in grass carp pituitary cells. [ABSTRACT FROM AUTHOR]

Neuropeptides, the largest amount of signaling molecules mainly secreted in the brain, play important roles in many physiological processes, including reproduction, growth, homeostasis and metabolism. Generally, neuropeptides exert physiological function via activating their specific G protein-coupled receptors (GPCRs). However, compared to mammals, the global information on neuropeptides and their GPCRs in teleost is still highly scarce. In this study, using grass carp as a model, a total of 91 preproneuropeptides of 23 families were identified from grass carp genome and transcriptome databases, as well as 112 GPCRs potentially binding with these neuropeptides. Then, using RNA-seq, the transcript levels of these preproneuropeptides and their GPCRs were examined among six different brain regions (hypothalamus, olfactory bulb, telencephalon, optic tectum, cerebellum, and medulla oblongata) and pituitary, respectively. Finally, basing on the tissue distribution of these preproneuropeptides and their receptors in different brain areas and pituitary, we further predict the potential function for each neuropeptide family in grass carp. This study will provide a significant contribution to further clarifying the functional role of each neuropeptide family in the neuroendocrine system of grass carp. • Global view of the biological actions of brain neuropeptides in fishes • Sequence analysis for 91 preproneuropeptides in grass carp • Transcript distribution for 91 preproneuropeptides and their 112 GPCRs in various brain areas and pituitary of grass carp. [ABSTRACT FROM AUTHOR]

Epidermal growth factor (EGF) is a member of the EGF-like ligands family, which plays a vital role in cell proliferation, differentiation, and folliculogenesis through binding with EGF receptors, including ErbB1 (EGFR/HER1), ErbB2 (HER2), ErbB3 (HER3), and ErbB4 (HER4). In mammals, many functional roles of EGF have been reported in the ovaries and breasts. However, little is known about the functions of EGF in the pituitary, especially in teleost. In this study, using grass carp pituitary cells as the model, we try to examine the direct pituitary actions of EGF in teleost. Firstly, transcriptomic analysis showed that 599 different expressed genes (DEGs) between the control and EGF-treatment group were mainly involved in cell proliferation, cell migration, signal transduction, and transcriptional regulation. Then, we further confirmed that EGF could significantly induce UTS1, EGR1, and MMP13 mRNA expression in a time-and dose-dependent manner. The stimulatory actions of EGF on UTS1 and EGR1 mRNA expression were mediated by the MEK1/2/ERK1/2 and PI3K/AKT/mTOR pathways coupled with both ErbB1 and ErbB2 in grass carp pituitary cells. The receptor specificity and signal transductions for the corresponding responses on MMP13 mRNA expression were also similar, except that the ErbB2 and PI3K/AKT/mTOR pathway were not involved. As we know, MMP13 could release EGF from HB-EGF. Interestingly, our data also showed that the MMPs inhibitor BB94 could suppress EGF-induced UTS1 and EGR1 mRNA expression. These results, taken together, suggest that the stimulatory actions of EGF on UTS1 and EGR1 mRNA expression could be enhanced by EGF-induced MMP13 expression in the pituitary. [ABSTRACT FROM AUTHOR]

In teleost, pigment in the skin and scales played important roles in various biological processes. Iridophores, one of the main pigment cells in teleost, could produce silver pigments to reflect light. However, the specific mechanism of the formation of silver pigments is still unclear. In our previous study, some transparent mutant individuals were found in the carp–goldfish nucleocytoplasmic hybrid (CyCa hybrid) population. In the present study, using transparent mutants (TM) and wild type (WT) of the CyCa hybrid as a model, firstly, microscopic observations showed that the silver pigments and melanin were both lost in the scales of transparent mutants compared to that in wild types. Secondly, genetic study demonstrated that the transparent trait in the CyCa hybrid was recessively inherent, and controlled by an allele in line with Mendelism. Thirdly, RNA-Seq analysis showed that differential expression genes (DEGs) between wild type and transparent mutants were mainly enriched in the metabolism of guanine, such as hydrolase, guanyl nucleotide binding, guanyl ribonucleotide binding, and GTPase activity. Among the DEGs, purine nucleoside phosphorylase 4a (pnp4a) and endothelin receptor B (ednrb) were more highly expressed in the wild type compared to the transparent mutant (p < 0.05). Finally, miRNA-Seq analysis showed that miRNA-146a and miR-153b were both more highly expressed in the transparent mutant compared to that in wild type (p < 0.05). Interaction analysis between miRNAs and mRNAs indicated that miRNA-146a was associated with six DEGs (MGAT5B, MFAP4, GP2, htt, Sema6b, Obscn) that might be involved in silver pigmentation. [ABSTRACT FROM AUTHOR]

Epidermal growth factor (EGF) is a potent regulator of cell function in many cell types. In mammals, the EGF/EGFR system played an important role in both pituitary physiology and pathology. However, it is not clear about the pituitary action of EGF in lower vertebrates. In this study, using grass carp as a model, we found that EGF could stimulate NK3R mRNA and protein expression through pituitary ErbB1 and ErbB2 coupled to MEK/ERK and PI3K/Akt/mTOR pathways. In addition, EGF could also induce pituitary somatolactin α (SLα) secretion and mRNA expression in a dose- and time-dependent manner in vivo and in vitro. The stimulatory actions of EGF on SLα mRNA expression were also mediated by PI3K/Akt/mTOR and MEK/ERK pathways coupled to ErbB1 and ErbB2 activation. Our previous study has reported that neurokinin B (NKB) could also induce SLα secretion and mRNA expression in carp pituitary cells. In the present study, interestingly, we found that EGF could significantly enhance NKB-induced SLα mRNA expression. Further studies found that NK3R antagonist SB222200 could block EGF-induced SLα mRNA expression, indicating an NK3R requirement. Furthermore, cAMP/PKA inhibitors and PLC/PKC inhibitors could both abolish EGF- and EGF+NKB-induced SLα mRNA expression, which further supported that EGF-induced SLα mRNA expression is NK3R dependent. [ABSTRACT FROM AUTHOR]