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1.
Br Poult Sci ; 60(3): 202-208, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-30968708

RESUMO

1. The slow skeletal muscle troponin I (TNNI1) gene has been found to be specifically expressed in slow muscle fibres and plays an important role in muscle development. The aim of this study was to determine the active control area of duck TNNI1 and identify the potential cis-regulatory elements in the promoter. 2. In this study, the TNNI1 promoter was first cloned by genome walking and the sequences were analysed using bioinformatics software. Firefly luciferase reporter gene vectors, driven by a series of constructs with progressive deletions, were used to identify the core transcriptional regulatory region of the duck TNNI1 gene. The methylation status of the CpG island in the TNNI1 promoter was detected in skeletal muscle on embryonic days 21 and 27, by bisulphite sequencing PCR (BSP). 3. The results showed two CpG islands presented in the promoter region, with one of the CpG islands located in the core transcriptional regulatory region (-2078/-885 bp). The total methylation levels of the 14 CpG sites were not altered between breast and leg muscles on embryonic days 21 and 27. However, four CpG sites (loci of positions 4, 11, 13, and 14) showed dramatically different methylation levels between breast and leg muscles at embryonic days 21 and 27. Analysis showed that multiple CpG sites had a significant correlation between the methylation levels of the CpG sites and mRNA expressions in skeletal muscle. Multiple transcription factor binding sites including Sp1, c-Myc, Oct-1 and NF-kB motifs were identified and might be responsible for transcriptional regulation of the TNNI1 gene. 4. These findings contribute to further understanding of the fundamental mechanism for transcriptional regulation of the TNNI1 gene in ducks.


Assuntos
Proteínas Aviárias/genética , Metilação de DNA , Patos/genética , Regulação da Expressão Gênica , Músculo Esquelético/metabolismo , Troponina I/genética , Animais , Proteínas Aviárias/metabolismo , Sequência de Bases , Ilhas de CpG , Patos/metabolismo , Regiões Promotoras Genéticas , Troponina I/metabolismo
2.
Anim Genet ; 50(3): 287-292, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-30994195

RESUMO

Plasma cholinesterase (PCHE) activity is an important auxiliary test in human clinical medicine. It can distinguish liver diseases from non-liver diseases and help detect organophosphorus poisoning. Animal experiments have confirmed that PCHE activity is associated with obesity and hypertension and changes with physiological changes in an animal's body. The objective of this study was to locate the genetic loci responsible for PCHE activity variation in ducks. PCHE activity of Pekin duck × mallard F2 ducks at 3 and 8 weeks of age were analyzed, and genome-wide association studies were conducted. A region of about 1.5 Mb (21.8-23.3 Mb) on duck chromosome 9 was found to be associated with PCHE activity at both 3 and 8 weeks of age. The top SNP, g.22643979C>T in the butyrylcholinesterase (BCHE) gene, was most highly associated with PCHE activity at 3 weeks (-logP = 21.45) and 8 weeks (-logP = 27.60) of age. For the top SNP, the strong associations of CC and CT genotypes with low PCHE activity and the TT genotype with high PCHE activity indicates the dominant inheritance of low PCHE activity. Problems with block inheritance or linkage exist in this region. This study supports that BCHE is a functional gene for determining PCHE levels in ducks and that the genetic variations around this gene can cause phenotypic variations of PCHE activity.


Assuntos
Colinesterases/genética , Patos/genética , Animais , Proteínas Aviárias/sangue , Proteínas Aviárias/genética , Proteínas Aviárias/metabolismo , Butirilcolinesterase/genética , Colinesterases/sangue , Colinesterases/metabolismo , Cruzamentos Genéticos , Patos/sangue , Patos/metabolismo , Feminino , Estudos de Associação Genética , Masculino , Polimorfismo de Nucleotídeo Único
3.
Theriogenology ; 131: 146-152, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-30965207

RESUMO

Lead (Pb) is an environmental pollutant and has toxic effect on birds. Selenium (Se) has alleviative effect on Pb poisoning. This study investigated mitigative effect of Se on autophagy in Pb-treated chicken testes. Seven-day-old male chickens were randomly divided into four groups with 45 birds in each group. The birds of the control group were offered drinking water (DW) and commercial diet (CD) (0.49 mg/kg Se). The birds of the Se group were offered DW and CD containing sodium selenite (SeCD) (1.00 mg/kg Se). The birds of the Pb group were offered DW containing lead acetate (PbDW) (350.00 mg/L Pb) and CD. The birds of the Pb/Se group were offered PbDW and SeCD. On the 30th, 60th, and 90th days, respectively; histology, antioxidant indexes (hydrogen peroxide (H2O2), catalase (CAT), total antioxidant capacity (TAOC), reduced glutathione (GSH), and superoxide dismutase (SOD)), mRNA and protein levels of autophagy-related genes (autophagy-related proteins 5, Beclin 1, Dynein, light chain 3 (LC3)-I, LC3-II, and mammalian target of rapamycin (mTOR)) were performed in chicken testes. The results of this study showed that Pb caused histological changes; increased H2O2 content; decreased CAT, TAOC, and SOD activities and GSH content; and increased mRNA and protein levels of the above autophagy-related genes except that mTOR decreased in chicken testes. Se alleviated the above changes. Se alleviated histological damage, oxidative stress, and autophagy in the Pb-treated chicken testes.


Assuntos
Autofagia/efeitos dos fármacos , Galinhas , Poluentes Ambientais/toxicidade , Chumbo/toxicidade , Estresse Oxidativo/efeitos dos fármacos , Selênio/farmacologia , Testículo/efeitos dos fármacos , Animais , Proteínas Aviárias/genética , Proteínas Aviárias/metabolismo , Biomarcadores/metabolismo , Masculino , RNA Mensageiro/metabolismo , Distribuição Aleatória
4.
J Therm Biol ; 81: 1-11, 2019 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-30975405

RESUMO

Heat-stress exposure increased the expression of heat-shock proteins (HSPs), B-cell lymphoma 2 (BCL-2) and anti-oxidative enzymes to maintain normal cellular function by attenuating the oxidative reaction and apoptosis. Reducing the stress response or enhancing anti-stress capability is an important goal in animal production. Our previous study indicated a protective role of flavangenol, a pine bark extract, in chicks after three hours of high-temperature exposure. However, the cellular mechanism of flavangenol was not clarified ex vivo. In the current study, we investigated the effect of flavangenol on cellular apoptosis and oxidation in heat-stressed treated chick brain cells (mixed neurons and glia cells). The primary brain cells were isolated from the diencephalon of 14-day-old chicks and cultured at 41.5 °C (to mimic the body temperature of young chicks), and were treated with flavangenol from day 3 of isolation to day 8. Cells were kept bathed in the cell culture dish under a high temperature (HT: 45 °C, 20 or 60 min) on day 8 and were then collected for analysis of cell viability as well as for HSP and other related gene expression. Flavangenol treatment significantly increased cell viability and BCL-2 mRNA expression, and attenuated HSP-70 and BCL-2-associated X protein mRNA expression. Moreover, flavangenol treatment elevated the mRNA expression of glutathione peroxidase in the HT group, which indicates that cellular anti-oxidative ability was strengthened by flavangenol. In conclusion, flavangenol may play a protective role in cells damaged or killed by heat stress by increasing cellular anti-oxidative pathways.


Assuntos
Antioxidantes/administração & dosagem , Apoptose/efeitos dos fármacos , Biflavonoides/administração & dosagem , Encéfalo/efeitos dos fármacos , Galinhas/genética , Regulação da Expressão Gênica/efeitos dos fármacos , Resposta ao Choque Térmico/efeitos dos fármacos , Fármacos Neuroprotetores/administração & dosagem , Proantocianidinas/administração & dosagem , Fenômenos Fisiológicos da Nutrição Animal , Animais , Proteínas Reguladoras de Apoptose/metabolismo , Proteínas Aviárias/metabolismo , Encéfalo/metabolismo , Sobrevivência Celular/efeitos dos fármacos , Galinhas/metabolismo , Proteínas de Choque Térmico/metabolismo , Resposta ao Choque Térmico/genética , Temperatura Alta , Masculino , Cultura Primária de Células , RNA Mensageiro/metabolismo
5.
Res Vet Sci ; 124: 256-262, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-30999161

RESUMO

Interferon-induced proteins with tetratricopeptide repeats (IFITs) are a family of proteins strongly induced downstream of type I interferon signaling. The function of IFITs has been investigated extensively in mammals. IFIT5 is the sole protein in this family found in birds and little information is available about the function of avian IFIT5. In this study, duck IFIT5 was cloned from peripheral blood mononuclear cells. Multiple amino acid sequence alignment and phylogenetic analysis showed that duck IFIT5 is highly homologous to chicken IFIT5. Tissue specificity analysis demonstrated that duck IFIT5 was ubiquitously expressed in all examined tissues of five-day-old ducklings, with the highest expression levels in heart, followed by thymus, cerebrum, liver, and lung; kidney expressed the lowest. Quantitative real-time PCR (qRT-PCR) analysis revealed that duck IFIT5 expression rapidly increased both in vitro and in vivo after stimulation with polyinosinic:polycytidylic acid [poly (I:C)] and infection with virulent duck hepatitis A virus type 3 (DHAV-3), respectively. Altogether, these results indicate that the expression of duck IFIT5 is positively correlated with viral load and may play an important role in the immune response to DHAV-3 infection. This study lays a foundation for further research into the innate antiviral immune responses of ducklings.


Assuntos
Patos/genética , Patos/imunologia , Proteínas de Neoplasias/genética , Sequência de Aminoácidos , Animais , Animais Recém-Nascidos/genética , Animais Recém-Nascidos/imunologia , Proteínas Aviárias/química , Proteínas Aviárias/genética , Proteínas Aviárias/metabolismo , Sequência de Bases , Clonagem Molecular , Vírus da Hepatite do Pato/fisiologia , Hepatite Viral Animal/imunologia , Proteínas de Neoplasias/química , Proteínas de Neoplasias/metabolismo , Fases de Leitura Aberta , Filogenia , Infecções por Picornaviridae/imunologia , Infecções por Picornaviridae/veterinária , Poli I-C/farmacologia , Doenças das Aves Domésticas/imunologia , Alinhamento de Sequência
6.
Mol Biotechnol ; 61(6): 400-409, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-30945164

RESUMO

Transgenic chickens are of great interest for the production of recombinant proteins in their eggs. However, the use of constitutive strong promoters or the tissue-specific ovalbumin promoter for the generation of the transgenic chickens have different drawbacks that have to be overcome in order to make chicken bioreactor an efficient production system. This prompted us to investigate the use of an alternative tissue-specific promoter, the vitellogenin promoter, which could overcome the difficulties currently found in the generation of chicken bioreactors. In the present work we establish and characterize a DNA construct consisting of a fragment of the 5´-flanking region of the chicken vitellogenin II gene cloned in a reporter vector. This construct is capable of showing the ability of the promoter to drive expression of a reporting gene in a tissue-specific manner and in a way that closely resembles physiologic regulation of vitellogenin, making it an ideal candidate to be used in the future for generation of avian bioreactors. Besides, we validate an in vitro culture system to test the performance of the DNA construct under study that could be used as a practical tool before generating any transgenic chicken. These results are important since they provide the proof of concept for the use of the vitellogenin promoter for future genetic modification of chickens bioreactors with improved characteristics in terms of quality of the recombinant protein produced.


Assuntos
Proteínas Aviárias/genética , Galinhas/genética , Vetores Genéticos/química , Proteínas Recombinantes de Fusão/genética , Vitelogeninas/genética , Região 5'-Flanqueadora , Animais , Animais Geneticamente Modificados , Proteínas Aviárias/metabolismo , Reatores Biológicos , Embrião de Galinha , Galinhas/metabolismo , Clonagem Molecular , Estradiol/farmacologia , Feminino , Fibroblastos/citologia , Fibroblastos/efeitos dos fármacos , Fibroblastos/metabolismo , Regulação da Expressão Gênica , Genes Reporter , Vetores Genéticos/metabolismo , Hepatócitos/citologia , Hepatócitos/efeitos dos fármacos , Hepatócitos/metabolismo , Luciferases/genética , Luciferases/metabolismo , Cultura Primária de Células , Regiões Promotoras Genéticas , Receptores Estrogênicos , Proteínas Recombinantes de Fusão/metabolismo , Transfecção/métodos , Vitelogeninas/metabolismo , Zigoto/efeitos dos fármacos , Zigoto/crescimento & desenvolvimento , Zigoto/metabolismo
7.
Int J Mol Sci ; 20(8)2019 Apr 22.
Artigo em Inglês | MEDLINE | ID: mdl-31013627

RESUMO

Overcoming P-glycoprotein (P-gp) efflux is a strategy to improve the absorption and pharmacokinetics of its substrate drugs. Berberine inhibits P-gp and thereby increases the bioavailability of the P-gp substrate digoxin in rodents. However, the effects of berberine on P-gp in chickens are still unclear. Here, we studied the role of berberine in modulating broilers P-gp expression and function through both in situ and in vitro models. In addition, molecular docking was applied to analyze the interactions of berberine with P-gp as well as with chicken xenobiotic receptor (CXR). The results showed that the mRNA expression levels of chicken P-gp and CXR decreased in the ileum following exposure to berberine. The absorption rate constant of rhodamine 123 increased after berberine treatment, as detected using an in situ single-pass intestinal perfusion model. Efflux ratios of P-gp substrates (tilmicosin, ciprofloxacin, clindamycin, ampicillin, and enrofloxacin) decreased and the apparent permeability coefficients increased after co-incubation with berberine in MDCK-chAbcb1 cell models. Bidirectional assay results showed that berberine could be transported by chicken P-gp with a transport ratio of 4.20, and this was attenuated by verapamil (an inhibitor of P-gp), which resulted in a ratio of 1.13. Molecular docking revealed that berberine could form favorable interactions with the binding pockets of both CXR and P-gp, with docking scores of -7.8 and -9.5 kcal/mol, respectively. These results indicate that berberine is a substrate of chicken P-gp and down-regulates P-gp expression in chicken tissues, thereby increasing the absorption of P-gp substrates. Our findings suggest that berberine increases the bioavailability of other drugs and that drug-drug interactions should be considered when it is co-administered with other P-gp substrates with narrow therapeutic windows.


Assuntos
Membro 1 da Subfamília B de Cassetes de Ligação de ATP/genética , Membro 1 da Subfamília B de Cassetes de Ligação de ATP/metabolismo , Berberina/farmacologia , Regulação da Expressão Gênica/efeitos dos fármacos , Membro 1 da Subfamília B de Cassetes de Ligação de ATP/química , Animais , Proteínas Aviárias/genética , Proteínas Aviárias/metabolismo , Berberina/química , Galinhas , Cães , Células Madin Darby de Rim Canino , Modelos Moleculares , Conformação Proteica , RNA Mensageiro/genética , Receptores Citoplasmáticos e Nucleares/genética , Receptores Citoplasmáticos e Nucleares/metabolismo , Relação Estrutura-Atividade
8.
Gen Comp Endocrinol ; 276: 69-76, 2019 05 15.
Artigo em Inglês | MEDLINE | ID: mdl-30851298

RESUMO

The function of oocyte-derived growth differentiation factor 9 (GDF9) in ovarian follicles has thus far been poorly defined in avian species compared with the defined function in mammals. Our aim here is to investigate the effects of GDF9 on steroidogenesis and on chicken ovarian granulosa cell (GC) mitosis. Primary GCs from both prehierarchical (6-8 mm in diameter, phGCs) and preovulatory follicles (F1-F5, poGCs) were cultured in the presence or absence of the GDF9 protein. The progesterone (P4) levels in the culture medium were then measured by radioimmunoassay (RIA), and the expression levels of steroidogenesis genes were detected by quantitative PCR. We found that GDF9 alone showed no significant effect on the P4 levels by regulating the expression of steroidogenesis genes, such as STAR, CYP11A1 and HSD3B. Further experiments indicated that GDF9 promoted follicle-stimulating hormone (FSH)-induced P4 production and STAR expression. GDF9 also rescued the FSH-induced decrease of FSH receptor (FSHR) expression but had no effect on the forskolin-induced P4, STAR and forskolin-inhibited FSHR expression levels, suggesting that GDF9 might achieve its regulatory role of P4 by enhancing FSHR and STAR expression. In addition, GDF9 also promoted GC cell cycle progression, regulated the gene transcription of related genes, potentiated DNA replication and inhibited apoptosis. Interestingly, these effects differed between the phGCs and the poGCs. To our knowledge, this is the first report that illustrates the function of GDF9 on chicken GCs and the effects on ovarian steroidogenesis. Our findings highlight the regulation of central oocytes on the surrounding granulosa cells and emphasize the interaction between paracrine signals and endocrine hormones on ovarian progesterone production; these findings contribute to the understanding of the development of avian ovarian follicles.


Assuntos
Galinhas/metabolismo , Hormônio Foliculoestimulante/farmacologia , Células da Granulosa/metabolismo , Fator 9 de Diferenciação de Crescimento/farmacologia , Progesterona/biossíntese , Animais , Apoptose/efeitos dos fármacos , Proteínas Aviárias/genética , Proteínas Aviárias/metabolismo , Ciclo Celular/efeitos dos fármacos , Células Cultivadas , Colforsina/farmacologia , DNA/biossíntese , Replicação do DNA/efeitos dos fármacos , Feminino , Regulação da Expressão Gênica/efeitos dos fármacos , Células da Granulosa/efeitos dos fármacos , Humanos , Ovulação/efeitos dos fármacos , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Radioimunoensaio
9.
Gene ; 701: 72-81, 2019 Jun 15.
Artigo em Inglês | MEDLINE | ID: mdl-30898701

RESUMO

Avian leukosis virus subgroup J (ALV-J) is an oncogenic retrovirus that causes severe economic losses to the poultry industry worldwide. Circular RNAs (circRNAs) are a class of non-coding RNAs that has been described in various biological systems and pathogenic processes. However, the immune mechanisms in response to circRNAs remain unknown. In this study, high-throughput transcriptome sequencing was used to detect circRNAs present in chicken macrophage (HD11) and chick embryo fibroblast (CEF) cells infected with ALV-J. We identified 7684 circRNAs from diverse genomic locations in CEF and HD11 after ALV-J infection, these RNAs showed complex expression patterns that differed based on the cells type and infection time. In total, 302 differentially expressed (DE) circRNAs and 164 DE circRNAs were identified in CEF and HD11 after ALV-J infection, respectively. CircRNA7419-associated with KDM4C- and circRNA6679 and circRNA6680-associated with TNFAIP6- were involved in the immune response upon ALV-J infection in CEF. Host genes were analyzed through further bioinformatics analysis. The result confirmed that a large number of DE circRNAs corresponded to several immune-associated or tumor-associated terms and pathways, such as Mucin type O-Glycan biosynthesis, MAPK signaling pathway, B cell receptor signaling, and Wnt signaling pathway in CEF, as well as Jak-STAT signaling pathway, apoptosis, and MAPK signaling pathway in HD11. CircRNAs related to the B cell receptor signaling pathway in CEF, and the Jak-STAT signaling pathway in HD11, were selected for circRNA-miRNA interaction network analyses. Our study indicates that circRNAs expression was altered by ALV-J infection in both CEF and HD11, and may play a key role in the progression of ALV-J infection.


Assuntos
Vírus da Leucose Aviária , Leucose Aviária , Galinhas , Sistema de Sinalização das MAP Quinases , Doenças das Aves Domésticas , RNA , Via de Sinalização Wnt , Animais , Leucose Aviária/genética , Leucose Aviária/metabolismo , Leucose Aviária/patologia , Vírus da Leucose Aviária/genética , Vírus da Leucose Aviária/metabolismo , Proteínas Aviárias/genética , Proteínas Aviárias/metabolismo , Linhagem Celular , Embrião de Galinha , Galinhas/genética , Galinhas/metabolismo , Galinhas/virologia , Doenças das Aves Domésticas/genética , Doenças das Aves Domésticas/metabolismo , Doenças das Aves Domésticas/patologia , Doenças das Aves Domésticas/virologia , RNA/genética , RNA/metabolismo
10.
Proc Natl Acad Sci U S A ; 116(14): 6766-6774, 2019 04 02.
Artigo em Inglês | MEDLINE | ID: mdl-30877242

RESUMO

Focal adhesion kinase (FAK) is a key signaling molecule regulating cell adhesion, migration, and survival. FAK localizes into focal adhesion complexes formed at the cytoplasmic side of cell attachment to the ECM and is activated after force generation via actomyosin fibers attached to this complex. The mechanism of translating mechanical force into a biochemical signal is not understood, and it is not clear whether FAK is activated directly by force or downstream to the force signal. We use experimental and computational single-molecule force spectroscopy to probe the mechanical properties of FAK and examine whether force can trigger activation by inducing conformational changes in FAK. By comparison with an open and active mutant of FAK, we are able to assign mechanoactivation to an initial rupture event in the low-force range. This activation event occurs before FAK unfolding at forces within the native range in focal adhesions. We are also able to assign all subsequent peaks in the force landscape to partial unfolding of FAK modules. We show that binding of ATP stabilizes the kinase domain, thereby altering the unfolding hierarchy. Using all-atom molecular dynamics simulations, we identify intermediates along the unfolding pathway, which provide buffering to allow extension of FAK in focal adhesions without compromising functionality. Our findings strongly support that forces in focal adhesions applied to FAK via known interactions can induce conformational changes, which in turn, trigger focal adhesion signaling.


Assuntos
Trifosfato de Adenosina/química , Proteínas Aviárias/química , Proteína-Tirosina Quinases de Adesão Focal/química , Simulação de Dinâmica Molecular , Desdobramento de Proteína , Trifosfato de Adenosina/metabolismo , Animais , Proteínas Aviárias/genética , Proteínas Aviárias/metabolismo , Galinhas , Ativação Enzimática , Proteína-Tirosina Quinases de Adesão Focal/genética , Proteína-Tirosina Quinases de Adesão Focal/metabolismo , Adesões Focais/enzimologia , Adesões Focais/genética , Mecanotransdução Celular/genética , Domínios Proteicos , Relação Estrutura-Atividade
11.
Proc Natl Acad Sci U S A ; 116(14): 6884-6890, 2019 04 02.
Artigo em Inglês | MEDLINE | ID: mdl-30886106

RESUMO

Animal skin pigment patterns are excellent models to study the mechanism of biological self-organization. Theoretical approaches developed mathematical models of pigment patterning and molecular genetics have brought progress; however, the responsible cellular mechanism is not fully understood. One long unsolved controversy is whether the patterning information is autonomously determined by melanocytes or nonautonomously determined from the environment. Here, we transplanted purified melanocytes and demonstrated that melanocytes could form periodic pigment patterns cell autonomously. Results of heterospecific transplantation among quail strains are consistent with this finding. Further, we observe that developing melanocytes directly connect with each other via filopodia to form a network in culture and in vivo. This melanocyte network is reminiscent of zebrafish pigment cell networks, where connexin is implicated in stripe formation via genetic studies. Indeed, we found connexin40 (cx40) present on developing melanocytes in birds. Stripe patterns can form in quail skin explant cultures. Several calcium channel modulators can enhance or suppress pigmentation globally, but a gap junction inhibitor can change stripe patterning. Most interestingly, in ovo, misexpression of dominant negative cx40 expands the black region, while overexpression of cx40 expands the yellow region. Subsequently, melanocytes instruct adjacent dermal cells to express agouti signaling protein (ASIP), the regulatory factor for pigment switching, which promotes pheomelanin production. Thus, we demonstrate Japanese quail melanocytes have an autonomous periodic patterning role during body pigment stripe formation. We also show dermal agouti stripes and how the coupling of melanocytes with dermal cells may confer stable and distinct pigment stripe patterns.


Assuntos
Galinhas/metabolismo , Codorniz/metabolismo , Pigmentação da Pele/fisiologia , Pele/metabolismo , Animais , Proteínas Aviárias/metabolismo , Embrião de Galinha , Conexinas/metabolismo , Melanócitos/citologia , Pele/citologia
12.
Artigo em Inglês | MEDLINE | ID: mdl-30790719

RESUMO

Adenosine monophosphate-activated protein kinase (AMPK) plays a pivotal role in the regulation of carbohydrate, lipid, and protein metabolism in animals. In this study, we examined whether any cross talk exists between glucocorticoids and AMPK in the regulation of the liver bile acid biosynthesis pathway. Dexamethasone treatment decreased the growth performance of broiler chickens. The liver mRNA levels of fatty acid transport protein (FATP-1), farnesoid X receptor (FXR), AMPK alpha 1 subunit (AMPKα1), and glucocorticoid receptor were significantly upregulated in DEX-treated broilers; the gene expression of liver cholesterol 7 alpha-hydroxylase (CYP7A1) was significantly downregulated. The protein level of liver CYP7A1 was significantly decreased by DEX treatment at both 24 and 72 h, while the protein level of p-AMPK/ t-AMPK stayed unchanged. In the in vitro cultured hepatocytes, compound C pretreatment blocked the increase in CYP7A1 protein level by DEX and significantly suppressed FATP-1, SREBP-1c, FXR, and CYP7A1 gene expression stimulated by DEX. Compound C treatment significantly reduces the protein level of p-AMPK, and the combination of compound C and DEX significantly reduces the protein level of t-AMPK. Thus, glucocorticoids affected liver AMPK and the bile acid synthesis signal pathway, and AMPK might be involved in the glucocorticoid effect of liver bile acid synthesis.


Assuntos
Proteínas Quinases Ativadas por AMP/metabolismo , Proteínas Aviárias/metabolismo , Galinhas/metabolismo , Dexametasona/farmacologia , Regulação da Expressão Gênica/efeitos dos fármacos , Glucocorticoides/farmacologia , Metabolismo dos Lipídeos/efeitos dos fármacos , Animais , Ácidos e Sais Biliares/biossíntese , Transdução de Sinais/efeitos dos fármacos
13.
Environ Sci Pollut Res Int ; 26(11): 10529-10536, 2019 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-30767103

RESUMO

Ammonia (NH3) is a highly irritant, alkaline gas. Atmospheric emission of NH3 was recognized as an environmental challenge. As a global issue, the NH3 emission survey with spatially detailed information demonstrated that the sources of atmospheric NH3 include agriculture (livestock wastes, fertilizers) and some industrial activities. As an environmental pollution, excessive NH3 exposure can induce many bird dysfunction. Neutrophils respond to multiple invading pathogens through different mechanisms. In order to investigate the effect of NH3 exposure on broilers' neutrophil, 1-day-old broilers were treated with/without NH3 for 28 days. We extracted neutrophils from peripheral blood of chicken with/without NH3 exposure and subsequently stimulated with PMA. Changes of cytokines and inflammatory bodies, heat shock proteins (HSPs), and glucose metabolism of neutrophil were examined in both cases. We not only explored that the index associated with inflammation changed due to NH3 exposure but also observed the status of neutrophils which was treated with PMA stimulation. After NH3 exposure, IL-1ß and IL-6 were significantly increased on broilers neutrophil. Inflammatory-related factors (NLRP3, ASC, and caspase-1) were significantly elevated. The mRNA expression of HSP70 and HSP90 was increased significantly. All glucose metabolism indicators were reduced. In summary, we concluded that NH3 enhanced inflammation and disrupted glucose metabolism, and increased the expression of HSPs and inflammatory factors. In addition, the sensitivity of neutrophils to exogenous stimuli was diminished. This information can not only be used to evaluate the damage of NH3-spiked neutrophils to chickens, but also provide clues for human health pathophysiology caused by excess NH3, providing valuable information for NH3 risk management.


Assuntos
Amônia/toxicidade , Proteínas Aviárias/genética , Citocinas/metabolismo , Glucose/metabolismo , Proteínas de Choque Térmico/genética , Inflamação/veterinária , Neutrófilos/efeitos dos fármacos , Doenças das Aves Domésticas/genética , Animais , Proteínas Aviárias/metabolismo , Galinhas , Citocinas/genética , Feminino , Fertilizantes/análise , Fertilizantes/toxicidade , Transtornos do Metabolismo de Glucose/metabolismo , Proteínas de Choque Térmico HSP70/metabolismo , Proteínas de Choque Térmico/metabolismo , Inflamação/etiologia , Inflamação/genética , Inflamação/metabolismo , Interleucina-1beta/genética , Interleucina-1beta/metabolismo , Interleucina-6/genética , Interleucina-6/metabolismo , Masculino , Neutrófilos/metabolismo , Doenças das Aves Domésticas/etiologia , Doenças das Aves Domésticas/imunologia , Doenças das Aves Domésticas/metabolismo
14.
Sci Total Environ ; 663: 380-386, 2019 May 01.
Artigo em Inglês | MEDLINE | ID: mdl-30716628

RESUMO

Hydrogen sulfide (H2S) is a toxic gas and one of the air pollutants of great concern. High-concentrated H2S can induce energy metabolism disturbance and apoptosis. However, the mechanism of H2S-induced liver injuries is unknown. Lipopolysaccharide (LPS), the main component of endotoxin, can cause fulminant hepatitis. Here, we evaluated the effects of H2S combined with LPS on the energy metabolism and apoptosis pathway in the liver using a one-day-old chicken as a model. Our results showed that the expression levels of energy metabolism-related genes (AMP-activated protein kinase (AMPK), Hypoxia-inducible factor-1 (HIF-1), aconitase 2 (ACO2), hexokinase1 (HK1), hexokinase 2 (HK2), lactate dehydrogenase A (LDHA), lactate dehydrogenase B (LDHB), phosphofructokinase (PFK), pyruvate kinase (PK) and succinate dehydrogenase B (SDHB)) tended to decrease, that the status of apoptosis increased, and that the expression levels of apoptosis-related genes (caspase3, BCL2, and bax) increased in H2S group, suggesting that H2S exposure disturbed the energy metabolism in the liver and induced hepatocyte apoptosis through the mitochondrial pathway. In addition, H2S combined with the LPS aggravated the level of energy metabolism disorders and apoptosis, indicating that H2S inhalation-induced energy metabolism disturbance is involved in LPS-mediated hepatocyte apoptosis through the mitochondrial pathway.


Assuntos
Apoptose/efeitos dos fármacos , Galinhas/metabolismo , Metabolismo Energético/efeitos dos fármacos , Sulfeto de Hidrogênio/efeitos adversos , Animais , Proteínas Aviárias/genética , Proteínas Aviárias/metabolismo , Galinhas/genética , Hepatócitos/efeitos dos fármacos , Hepatócitos/fisiologia , Exposição por Inalação/análise , Lipopolissacarídeos/farmacologia , Mitocôndrias/efeitos dos fármacos , Mitocôndrias/metabolismo , RNA Mensageiro/genética , RNA Mensageiro/metabolismo
15.
Br Poult Sci ; 60(3): 323-329, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-30784300

RESUMO

1. The objective of this study was to reveal the role of chicken RB1 (Gallus gallus RB1, gRB1) in the proliferation of preadipocytes. 2. To measure gene expression of gRB1 in the proliferation of chicken preadipocyte, quantitative real-time PCR was used. The expression levels of gRB1 transiently increased during this process. 3. To detect the effect of gRB1 on the proliferation of chicken preadipocyte, MTT assay and cell-cycle analysis were performed. MTT assay showed that overexpression of gRB1 significantly suppressed (P < 0.05) the proliferation of chicken preadipocytes, and knockdown of gRB1 promoted the proliferation of chicken preadipocytes. Cell-cycle analysis showed that the proportion of preadipocytes in the G1 and G2 phases significantly increased (P < 0.05), and the proportion of preadipocytes in the S phase significantly decreased (P < .05) after up-regulation of the expression of gRB1. The proportion of preadipocytes in the S phase significantly increased (P < 0.05) after down-regulation of gRB1. 4. Quantitative real-time PCR was used to detect the effect of gRB1 on the expression of genes related to proliferation of chicken preadipocytes. Gene expression analysis showed that gRB1 knockdown promoted markers indicating proliferation of Ki-67 (MKi67) expression at 96 h (P < 0.05), and overexpression of gRB1 reduced MKi67 expression at 72 h (P < 0.05). 5. This study demonstrated that gRB1 inhibited preadipocyte proliferation at least in part by inhibiting the G1 to S phase transition.


Assuntos
Adipócitos/fisiologia , Proteínas Aviárias/genética , Proliferação de Células/genética , Galinhas/fisiologia , Fator de Transcrição E2F1/genética , Animais , Proteínas Aviárias/metabolismo , Fator de Transcrição E2F1/metabolismo , Reação em Cadeia da Polimerase em Tempo Real
16.
Poult Sci ; 98(5): 2008-2013, 2019 May 01.
Artigo em Inglês | MEDLINE | ID: mdl-30597054

RESUMO

Salmonella enterica serovar Typhimurium (S. Typhimurium) is a primary avian pathogen responsible for severe intestinal pathology in younger chickens and economic losses to poultry industry. Furthermore, S. Typhimurium is also able to cause infection in humans, characterized by acute gastrointestinal disease. A study was conducted to investigate antibody response and expression kinetics of interferon gamma (IFNγ), interleukin (IL-12, and IL-18) genes in broiler chicken at 0, 1, 3, 5, 7, 9, 11, 13, and 15 D post infection following experimental infection of S. Typhimurium. Immunological studies showed higher titres of IgG and IgM in the infected group as compared to the age-matched un-infected control group. The Real-Time PCR-based gene expression analysis revealed significant increase of IFNγ, IL-12, and IL-18 mRNA levels in the infected group as compared to their respective controls (P < 0.05). The present study shall help in understanding the immune responses in birds, thus allowing development of more effective vaccines and vaccination strategies.


Assuntos
Galinhas , Doenças das Aves Domésticas/genética , Doenças das Aves Domésticas/imunologia , Salmonelose Animal/genética , Salmonelose Animal/imunologia , Salmonella typhimurium/fisiologia , Animais , Anticorpos Antibacterianos/sangue , Formação de Anticorpos , Proteínas Aviárias/genética , Proteínas Aviárias/metabolismo , Expressão Gênica , Perfilação da Expressão Gênica/veterinária , Interferon gama/genética , Interferon gama/metabolismo , Interleucina-12/genética , Interleucina-12/metabolismo , Interleucina-18/genética , Interleucina-18/metabolismo , Doenças das Aves Domésticas/microbiologia , RNA Mensageiro/genética , Salmonelose Animal/microbiologia
17.
Poult Sci ; 98(5): 2220-2230, 2019 May 01.
Artigo em Inglês | MEDLINE | ID: mdl-30597072

RESUMO

This study evaluated the effect of reduced dietary energy (ME) and crude protein (CP) levels along with inclusion of a phytogenic feed additive (PFA) on gut microbiota composition and gene expression of Toll-like receptor(s) (TLR), tight junction proteins, and inflammatory cytokines expressed in secondary lymphoid organs. Depending on dietary ME and CP level down regulation and the inclusion or not of PFA at 125 mg/kg diet, 450 one-day-old male broilers were allocated in the following 6 treatments for 42 D according to a 3 × 2 factorial design: A: diet formulated optimally to meet broiler nutrient requirements; APh: A+PFA; B: suboptimal in ME and CP levels by 3%; BPh: B+PFA; C: suboptimal in ME and CP levels by 6%; CPh: C+PFA. Diet type and PFA supplementation were shown to affect mostly the mucosa-associated microbiota compared to the luminal ones. Ileal mucosa-associated total bacteria (PD= 0.005), Lactobacillus spp. (PD= 0.003), and Clostridium cluster XIVa (PD= 0.009) were affected by diet type with broilers fed diet B having lower levels compared to broilers fed diets A or C. Moreover, diet type affected cecal mucosa-associated Lactobacillus spp. (PD= 0.002) with broilers fed diet C having lower levels compared to broilers fed diets A or B. Supplementation with PFA resulted in higher levels of cecal mucosa-associated Bacteroides (PP= 0.031), Clostridium cluster IV (PP= 0.007), and Clostridium cluster XIVa (PP= 0.039). Diet type affected TLR2 (PD= 0.046) and claudin 5 (PD= 0.027) in cecal epithelium. Lower TLR2 (PP= 0.021) and higher zonula occludens 2 (PP= 0.031) relative gene expressions were seen in ileal epithelium following PFA supplementation. Moreover, in cecal epithelium, PFA supplementation resulted in lower TLR2 (PP < 0.001) and higher zonula occludens 2 (PP= 0.009), claudin 5 (PP= 0.005) and occludin (PP= 0.039) relative gene expressions. There were no significant diet type and PFA effects on cytokines in secondary lymphoid organs, except for a dietary effect on transforming growth factor beta 4 (PD= 0.023) in cecal tonsils. In conclusion, PFA inclusion beneficially modulated elements of gut microbiota, Toll-like signaling molecules and gut tight junction genes.


Assuntos
Proteínas Aviárias/genética , Galinhas/genética , Galinhas/microbiologia , Suplementos Nutricionais/análise , Microbioma Gastrointestinal , Expressão Gênica , Ração Animal/análise , Animais , Proteínas Aviárias/metabolismo , Ceco/efeitos dos fármacos , Ceco/metabolismo , Citocinas/genética , Citocinas/metabolismo , Dieta/veterinária , Microbioma Gastrointestinal/efeitos dos fármacos , Expressão Gênica/genética , Íleo/efeitos dos fármacos , Íleo/metabolismo , Masculino , Distribuição Aleatória , Baço/efeitos dos fármacos , Baço/metabolismo , Proteínas de Junções Íntimas/genética , Proteínas de Junções Íntimas/metabolismo , Receptores Toll-Like/genética , Receptores Toll-Like/metabolismo
18.
Poult Sci ; 98(5): 2201-2210, 2019 May 01.
Artigo em Inglês | MEDLINE | ID: mdl-30608557

RESUMO

In mammals, the AMP-activated protein kinase (AMPK) pathways in the central and peripheral tissues coordinately integrate inputs from multiple sources to regulate energy balance. To investigate the effects of the fatty liver hemorrhagic syndrome (FLHS) caused by high-energy, low-protein diets and to explore the potential role of AMPK in the energy homeostasis of FLHS, 60 laying hens were equally divided into 2 groups: control group (basal diet) and experimental group (high-energy, low-protein diet). Liver tissues were subjected to histopathological analysis. Liver tissues were also collected on the 100th day to determine the levels of total cholesterol, triglyceride (TG), high-density lipoprotein cholesterol (HDL-Ch), low-density lipoprotein cholesterol (LDL-Ch), aspartate aminotransferase, and alanine aminotransferase in plasma. Additionally, the mRNA expression levels of AMPK signaling pathway related genes in liver were determined by quantitative RT-PCR. The results showed that histopathological lesions presented different degrees of lipid vacuolization in hepatocytes. In combination with hematoxylin and eosin and oil red O staining, the experimental group was divided into mild group and severe group. In the severe group, contents of TG and LDL-Ch were extremely significantly increased (P < 0.01) compared to the control group, and HDL-Ch content was extremely significantly decreased (P < 0.01). The serine-threonine kinase 11 and AMPKα1 mRNA expression levels were downregulated, while acetyl-CoA carboxylase, fatty acid synthase, hepatocyte nuclear factor-4α, 3-hydroxy-3-methyl glutaryl coenzyme A reductase and carnitine palmitoyltransferase-I mRNA expression levels were upregulated by a high-energy and low-protein diet. Taken together, these findings suggest that a functional AMPK signaling pathway exists in chickens and AMPK may alter the energy balance in the FLHS induced by high-energy, low-protein diets.


Assuntos
Proteínas Quinases Ativadas por AMP/genética , Galinhas , Dieta com Restrição de Proteínas/veterinária , Fígado Gorduroso/veterinária , Hemorragia/veterinária , Doenças das Aves Domésticas/genética , Transdução de Sinais , Proteínas Quinases Ativadas por AMP/metabolismo , Ração Animal/análise , Animais , Proteínas Aviárias/genética , Proteínas Aviárias/metabolismo , Fígado Gorduroso/genética , Feminino , Hemorragia/genética
19.
Poult Sci ; 98(5): 2231-2240, 2019 May 01.
Artigo em Inglês | MEDLINE | ID: mdl-30624755

RESUMO

We examined the effects of dynamic feeding low- and high-methionine diets on the transcriptional level of glucose and lipid metabolism-related genes in the liver of laying hens. A total of 180 laying hens (Brown Hy-line, 41 wk old) were exposed to 16 h of light and 8 h of darkness with lights on at 06:00 h local time (Zeitgeber time [ZT] 0) and allocated into 3 equal groups with 6 replicates. The control group was fed a 0.30% methionine diet. The low-high (LH) group was fed 0.27% methionine diet at ZT1.5 and 0.33% methionine diet at ZT8.5. The feeding regime in high-low (HL) group was opposite to that of the LH group. After 10 wk, a total of 108 hens (6 hens/group/time point) were randomly and equally selected for further analysis. Blood and liver tissues were collected at ZT21.5, ZT1.5, ZT5.5, ZT9.5, ZT13.5, and ZT17.5. The results showed that the serum ALP activity in the HL group was the highest at ZT9.5 and ZT13.5, and that the serum glucose content in the HL group was lowest at ZT21.5 compared to the other 2 groups (P < 0.05). The mesors, amplitudes, and acrophases of the cosine curves for the CLOCK, BMAL1, CRY1, PER2, and PER3 genes were altered in the LH and HL groups. G6PC3, PCK2, GPAT, INSIG2, FASN, ACACA, ACOX1, HMGCR, LDLR, and PPARA expression in the liver were affected by the feeding regime at some time point (P < 0.05). Two-way analysis of variance comparisons showed some significant effects of time on the mRNA expression of G6PC3, G6PC2, PCK2, FBP1, FBP2, GCK, GYS2, FASN, GPAT, HMGCR, LDLR, ACC, SREBP2, and INSIG1 (P < 0.05). Moreover, different feeding regimes significantly affected the expression of FASN, GPAT, HMGCR, LDLR, ACACA, SREBP1, and INSIG1 (P < 0.05). In conclusion, dynamical feeding low- and high-methionine diets affected the variation of serum ALP and glucose levels, as well as the mRNA expression of circadian clock- and glucose and lipid metabolism-related genes in the liver of laying hens.


Assuntos
Proteínas Aviárias/genética , Galinhas/genética , Relógios Circadianos/genética , Expressão Gênica , Glucose/metabolismo , Metabolismo dos Lipídeos/genética , Metionina/metabolismo , Ração Animal/análise , Animais , Proteínas Aviárias/metabolismo , Galinhas/metabolismo , Relógios Circadianos/efeitos dos fármacos , Dieta/veterinária , Relação Dose-Resposta a Droga , Feminino , Expressão Gênica/efeitos dos fármacos , Metabolismo dos Lipídeos/efeitos dos fármacos , Fígado/efeitos dos fármacos , Fígado/metabolismo , Metionina/administração & dosagem , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Distribuição Aleatória
20.
Poult Sci ; 98(5): 2272-2280, 2019 May 01.
Artigo em Inglês | MEDLINE | ID: mdl-30624759

RESUMO

Within the last 60 yr genetics of broilers have changed to produce rapid growing birds that achieve market weight in 6 wk or less. To investigate the differences in factors that play a role in nutrient processing and uptake between modern fast growing (Ross) and slow growing broilers not selected for growth (ACRBC), a study was carried comparing the expression of 13 genes that encode amino acid transporters (ASCT1, ATBo,+, BoAT, bo, +AT, CAT1, CAT2, EAAT3, γ+LAT1, and LAT1) and sugar transporters (GLUT2 and GLUT5), as well as aminopeptidase (APN) and the di- and tri-peptide transporter PepT1. The growth rate of Ross birds was approximately 4 times greater than that of ACRBCs, and the feed conversion ratio (FCR) was greater in ACRBCs at all-time points measured. Gene expression in the duodenum, jejunum, and ileum was measured at 1, 3, 5, 10, and 14 d post hatch (PH). The expression of genes that encode proteins (particularly ASCT1, ATBo, +, and BoAT) located at the brush border of the gut epithelium was generally higher in ACRBCs especially at earlier time points. The expression of genes that encode proteins located at the basolateral surface of the gut epithelium was less affected. The expression of GLUT2 and GLUT5 was significantly decreased in ACRBCs at most time points and gut segments. Based on the present data we conclude that expression of brush border and sugar transporters in the small intestine can be correlated with growth. Presented increases the identification of the factors that influence growth and will assist future studies of the function of these molecules.


Assuntos
Proteínas Aviárias/metabolismo , Galinhas/genética , Galinhas/metabolismo , Expressão Gênica , Nutrientes/metabolismo , Seleção Genética , Aminopeptidases/genética , Aminopeptidases/metabolismo , Animais , Proteínas Aviárias/genética , Masculino , Proteínas de Membrana Transportadoras/genética , Proteínas de Membrana Transportadoras/metabolismo , Transportador 1 de Peptídeos/genética , Transportador 1 de Peptídeos/metabolismo
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