RESUMO
Antibodies against the extracellular domains of the chicken leptin receptor were used to study the biological function of leptin in growing chickens. Both polyclonal and monoclonal anti-LEPR antibodies were administered intramuscularly to 30-d-old Chinese indigenous Gushi pullets. Both antibody preparations increased feed intake for 6â¯h after injection and reduced plasma concentrations of glucose, triglycerides, and both high- and low-density lipoproteins. The antibody treatments also upregulated agouti-related peptide and neuropeptide Y in the hypothalamus and downregulated proopiomelanocortin, melanocortin 4 receptor, and leptin receptor. The treatments also upregulated leptin receptor, acetyl CoA carboxylase beta, and acyl-CoA oxidase in the liver, abdominal fat, and breast muscle and downregulated sterol regulatory element-binding protein-1 and fatty acid synthase. Furthermore, even though the anti-leptin receptor antibodies failed to affect leptin receptor signaling transduction when administered alone, they did augment the induction of leptin receptor signaling transduction by leptin. These results demonstrate that antibodies against the extracellular domains of leptin-specific receptor enhance, but do not mimic, the ability of leptin to activate receptors. Furthermore, the enhanced leptin bioactivity observed after the intramuscular injection of anti-LEPR antibodies confirmed the occurrence of de novo leptin in the peripheral tissues and blood of treated chickens.
Assuntos
Galinhas , Receptores para Leptina/metabolismo , Animais , MasculinoRESUMO
BACKGROUND AND OBJECTIVE: Currently, there is no single golden standard for diagnosing ulcerative colitis (UC). Now serum αvß6 autoantibodies have shown promise as a diagnostic tool for UC. Here the aim was to determine the diagnostic performance of serum αvß6 autoantibodies for UC. METHODS: PubMed, the Cochrane Library, the Embase, and the Web of Science were searched comprehensively. STATA software was utilized to analyze the relevant data. RESULTS: 9 studies from 6 articles with 1827 subjects were eligible. The summary sensitivity and specificity of serum αvß6 autoantibodies to diagnose UC were 0.82 (95 % confidence interval (CI): 0.65-0.92) and 0.94 (95 % CI: 0.90-0.97) with an area under the summary receiver operating characteristic curve of 0.96 (95 % CI: 0.94-0.97). Subgroup analysis was conducted owning to substantial heterogeneity between studies (I2 = 97 % and P < 0.001). The aggregate sensitivity and specificity to diagnose UC in adults were 0.75 (95 % CI: 0.61-0.86) and 0.95 (95 % CI: 0.90-0.97), and when using a threshold of mean control+3SD, 0.80 (95 % CI: 0.60-0.91) and 0.96 (95 % CI: 0.90-0.99), respectively. Additionally, to differentiate UC from healthy participants, non-inflammatory bowel disease, and Crohn's disease, the overall specificity was 0.96, 0.88, and 0.80, respectively. CONCLUSIONS: serum αvß6 autoantibodies, as a non-invasive tool, demonstrated good diagnostic accuracy for UC. However, their application may be limited in some immune-related disorders, and further studies are needed for validation.
Assuntos
Autoanticorpos , Colite Ulcerativa , Humanos , Colite Ulcerativa/sangue , Colite Ulcerativa/diagnóstico , Colite Ulcerativa/imunologia , Autoanticorpos/sangue , Sensibilidade e EspecificidadeRESUMO
Growth hormone (GH) is required for normal postnatal development in poultry; however, no immunoassay exists to assess its levels in geese plasma, hindering the study of endocrine regulation in this species. We developed a sandwich ELISA to determine the GH concentrations in the plasma of geese. Recombinant goose GH was produced using a eukaryotic expression system and purified for use as the reference standard in ELISA and the antigen for producing the polyclonal antibodies in rabbits. Rabbit anti-goose GH polyclonal antibody was used to coat the wells of the ELISA plate, and its biotinylated form served as the detection antibody. An avidin-conjugated horseradish peroxidase was used to bind the detection antibody and catalyze the chromogenic reaction of 3,3,5,5-tetramethylbenzidine and H2O2. A sigmoidal curve was fitted to the optical density and the log of the standard GH concentration using the four-parameter logistic model. The sensitivity of the assay was less than 0.156 ng/mL. The intra- and interassay coefficients of variation were less than 9 and 13%, respectively. The response curve of the serially diluted plasma samples from geese exhibited a good parallel relationship with that observed for the reference standards. The assay effectively detected differences in GH concentrations in plasma samples from geese at various physiological stages; thus, it will be useful for future study of their growth and metabolism.
Assuntos
Gansos , Hormônio do Crescimento , Animais , Ensaio de Imunoadsorção Enzimática/veterinária , Gansos/sangue , Hormônio do Crescimento/sangue , Peróxido de Hidrogênio , CoelhosRESUMO
The aim of this study was to determine the cellular and molecular mechanisms of leptin (LEP) and the leptin receptor (LEPR) in testicular development of prepubertal ganders. In an in vivo animal experiment, active immunization against LEPR severely depressed prepubertal testicular development by significantly reducing testicular weights at 200 and 227 days of age. The number of elongated spermatids in the seminiferous tubules was also significantly decreased by immunization with LEPR at ages of 200 and 227 days. Inhibition of testicular development by LEPR immunization was associated with decreases in LHR, StAR, 3ß-HSD, CYP11A1, CYP17A1, and PRLR mRNA expression levels in testicular tissue, which resulted in a significant decrease in testosterone synthesis. In the in vitro experiments, the addition of LEP combined with anti-LEPR antibodies strengthened LEPR signal transduction, and inhibited significantly testosterone production in cultured Leydig cells isolated from prepubertal gander testes. The mRNA expression of LHR, StAR, 3ß-HSD, CYP11A1, CYP17A1 also decreased significantly after treatment with LEP combined with anti-LEPR antibodies in cultured Leydig cells. These results suggest that anti-LEPR antibodies strengthen LEPR signaling transduction in the presence of LEP, and immunization against LEPR inhibited testes development and testosterone secretion in prepubertal ganders.
Assuntos
Leptina , Receptores para Leptina , Animais , Masculino , Receptores para Leptina/genética , Transdução de Sinais , Testículo , TestosteronaRESUMO
A meta-analysis was performed to evaluate the effect of oestrogen receptor-alpha (ESR1) gene PvuII polymorphism on fracture risk. It included published data from relevant studies (up to May 2010) identified from Medline, Embase and Current Contents. The 13 included studies contained 1279 fracture cases and 6069 controls. The combined results based on these studies showed no relationship between ESR1 gene PvuII polymorphism and fracture risk. No significant difference in genotype distribution was found when stratifying by race. When stratifying by fracture type, it was found that vertebral fracture cases had a significantly higher frequency of the PvuII pp genotype than controls in five studies (552 cases and 2350 controls). This meta-analysis suggests a modest but statistically significant association between the ESR1 PvuII pp genotype and vertebral fracture.
Assuntos
Receptor alfa de Estrogênio/genética , Fraturas Ósseas/genética , Polimorfismo Genético/genética , Estudos de Casos e Controles , Humanos , Fatores de RiscoRESUMO
The aim of this study was to investigate the effects of active immunization against recombinant Anti-Müllerian hormone (AMH) protein on the ovarian follicular development, egg production, and molecular regulatory mechanisms in broody-prone Zhedong White geese. For this, a recombinant goose AMH protein was expressed using a prokaryotic expression system. Fifty incubating geese from the same genetic background were selected and equally divided into two groups. The immunization group was actively immunized against the recombinant goose AMH protein, whereas the control group was immunized against bovine serum albumin (BSA). Immunization against AMH accelerated ovarian follicular development and increased clutch sizes by one to two eggs in two consecutive laying-incubation cycles. Furthermore, immunization against AMH upregulated the mRNA transcription levels of the FSH-beta gene in the pituitary gland, and FSHR, 3beta-HSD, and Smad4 genes in the granulosa layer of pre-ovulatory follicles; however, immunization downregulated the expression of the OCLN gene in the granulosa layer of pre-ovulatory follicles, and Smad5 and Smad9 genes in the granulosa layer of SYFs. These results suggest that AMH might hinder ovarian follicular development by decreasing both pituitary FSH secretion as well as ovarian follicular sensitivity to FSH. The latter molecular mechanism could be fulfilled by regulating Smad5 or Smad9 signals in SYFs, as well as the FSHR and Smad4 signals that affect progesterone synthesis and yolk deposition in the pre-ovulatory follicles.
Assuntos
Anseriformes/fisiologia , Hormônio Antimülleriano/imunologia , Folículo Ovariano/imunologia , Ovulação/imunologia , Animais , Clonagem Molecular , Feminino , Hormônio Liberador de Gonadotropina/genética , Hormônio Liberador de Gonadotropina/metabolismo , Imunização , Folículo Ovariano/metabolismo , Ovulação/fisiologia , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Proteínas RecombinantesRESUMO
Photoperiodic control is essential for manipulating the reproductive performance of avian species. This study was conducted to assess the neuroendocrine mechanisms that regulate reproductive functions of Yangzhou geese when there are different monochromatic light colors from light emitter diode (LED) sources. A flock of geese was divided into four groups with white, red, blue, and green light treatments being imposed. The results indicated that peak laying rates and reproductive performance were greater in geese treated with white or red as compared with blue or green light treatments. The fertilization rate of eggs and hatchability of fertilized eggs were greater with the white or red as compared with blue or green light treatments. There was a greater abundance of OPN5, Dio2, c-Fos, and GnRH-I mRNA in the hypothalamus earlier in the treatment period and abundances of these hypothalamic factors were greater with the white or red light treatments. Abundances of pituitary LH beta and FSH beta mRNA increased at a lesser rate with the blue or green light treatments and were in greater abundances with the white or red light treatments. The lighting regimen also resulted in photo-refractoriness with there being greater abundances of GnIH, VIP, and PRL mRNA with the use of white or red light treatments. The results indicate that the use of white or red monochromatic lights while imposing a long photoperiod of 11 h daily could result in sustaining functions of the reproductive system of Yangzhou geese for considerably longer times, thus, resulting in greater egg-laying performance.
Assuntos
Cor , Gansos/fisiologia , Oviposição/fisiologia , Animais , Feminino , Estimulação Luminosa , Fotoperíodo , Distribuição AleatóriaRESUMO
Hungarian White geese are regarded as good producers of meat, eggs, and feathers, but specific lighting schedules are required to improve their egg-laying performance. This study reveals the neuroendocrine regulatory mechanisms that govern the reproductive activities and egg-laying performances of Hungarian White geese. The results indicated that increasing the daily photoperiod from a short 8â¯h period to either 11â¯h or 14â¯h initiated reproduction. Egg-laying rates increased faster in the 14â¯h group, peaking (48.2%) on day 33 as compared to the peak (52.67%) reached on day 53 in the 11â¯h group. Changes to the plasma estradiol and progesterone concentrations produced similar patterns in the two groups. In the hypothalamus, OPN5, Dio2, c-Fos, and GnRH-I expression levels showed similar sequential increases and decreases. Changes in GnIH and VIP expression levels were the opposite to those of GnRH-I, but the levels peaked earlier under the 14â¯h photoperiod conditions. Pituitary LH beta and FSH beta expression levels increased at slower rates but remained significantly higher in the 11â¯h group than in the 14â¯h group. However, pituitary PRL expression increased considerably earlier and was higher in 14â¯h geese than in 11â¯h geese, which was opposite to the observed egg-laying rate patterns. An increase from a short to a relatively long photoperiod (11â¯h) regulated the neuroendocrine system and led to reproductive activities being sustained for a longer period, which resulted in high egg-laying performances.
Assuntos
Anseriformes/fisiologia , Oviposição/fisiologia , Fotoperíodo , Animais , Peso Corporal , Estradiol/sangue , Feminino , Regulação da Expressão Gênica/efeitos da radiação , Sistema Hipotálamo-Hipofisário/fisiologia , Folículo Ovariano , Ovário/fisiologia , Progesterona/sangueRESUMO
The aim of this study was to determine the cellular and molecular mechanisms of prolactin (PRL) in testicular development of prepubertal cockerels. In an in vivo animal experiment, active immunization against PRL severely depressed prepubertal testicular development by significantly reducing testicular weights at both 122 and 164 d of age. The number of elongated spermatids in the seminiferous tubules was also significantly decreased by immunization with 199-residue chicken PRL (cPRL) at age 122 d. Inhibition of testicular development by cPRL immunization was associated with decreases in LH receptor (LHR), FSH receptor (FSHR), Stat5b, P450scc, steroidogenic acute regulatory (StAR) protein, and 3ß-hydroxysteroid dehydrogenase (3ß-HSD) mRNA expression levels in testicular tissue. In in vitro experiments, testosterone production by cultured Leydig cells isolated from prepubertal cockerel testes was dose-dependently enhanced by treatment with bioactive recombinant PRL, but a lesser response was seen with high concentrations of PRL. The distinct changes in testosterone production in response to high and low concentrations of added PRL were paralleled by similar patterns of change in the mRNA levels of Stat5b, LHR, P450scc, StAR, 3ß-HSD, and CYP17A1 in cultured Leydig cells, as well as protein amounts of phosphorylated Jak2 and Stat5a/b. In conclusion, low to medium doses of PRL potentiate testis development in prepubertal cockerels by enhancing testosterone secretion from Leydig cells via activation of PRLR/Stat5b signal transduction, which upregulates mRNA expression of LHR and testosterone synthesizing enzymes. However, this positive regulation was weaker in response to a high dose of PRL, which reduced PRLR/Stat5b signal transduction and the expression of genes involved in LH signaling and testosterone synthesis.
Assuntos
Galinhas/crescimento & desenvolvimento , Prolactina/farmacologia , Testículo/efeitos dos fármacos , Animais , Células Cultivadas , Regulação da Expressão Gênica/efeitos dos fármacos , Células Intersticiais do Testículo/efeitos dos fármacos , Masculino , Prolactina/imunologia , Testículo/citologia , Testículo/crescimento & desenvolvimentoRESUMO
Five single nucleotide polymorphisms (SNP) of the chicken insulin-like factor binding protein 2 (IGFBP2) gene were selected to genotype a F2 designed population with restriction fragment length polymorphisms and single stranded-conformation polymorphisms. The associations of the SNP and their haplotypes with chicken growth and carcass traits were analyzed. Results showed that the difference induced by the haplotypes derived from the 5 SNP was more significant than that by the single SNP in the genotype-phenotype association analysis. The haplotypes were associated with BW at hatch and at 21, 28, 42, 49, 56, and 90 d of age, as well as eviscerated weight with giblets (EWG), eviscerated weight (EW), and weights of heart, liver, and gizzard (HLGW) (P < or = 0.01). The haplotypes were also related to BW at 7, 14, and 35 d of age, breast depth, carcass weight, and breast muscle weight (P < or = 0.05). Significant and suggestive dominant effects of H1H5 diplotype were detected for BW at 7, 14, 21, 28, and 90 d of age, as well as breast depth, carcass weight, eviscerated weight with giblets, eviscerated weight, breast muscle weight, leg muscle weight, and weights of heart, liver, and gizzard. It was concluded that H1H5 was the most advantageous diplotype, and H4H10 was the negative diplotype for growth and carcass traits.
Assuntos
Galinhas/crescimento & desenvolvimento , Galinhas/genética , Proteína 2 de Ligação a Fator de Crescimento Semelhante à Insulina/genética , Animais , Sequência de Bases , Peso Corporal/genética , Ligação Genética , Haplótipos , Proteína 2 de Ligação a Fator de Crescimento Semelhante à Insulina/fisiologia , Polimorfismo de Nucleotídeo Único , Característica Quantitativa HerdávelRESUMO
In this study, immunization against chicken leptin receptor (cLEPR) extracellular domain (ECD) was applied to investigate leptin regulation and LEPR biofunction in growing chicken pullets. A recombinant protein (cLEPR ECD) based on the cLEPR complemenary DNA sequence corresponding to the 582nd to 796th amino acid residues of cLEPR mature peptide was prepared and used as antigen. Immunization against cLEPR ECD in growing chickens increased anti-cLEPR ECD antibody titers in blood, enhanced proportions of phosphorylated janus kinase 2 (JAK2) and served as signal transducer and activator of transcription 3 (STAT3) protein in liver tissue. Chicken live weight gain and abdominal fat mass were significantly decreased (P < 0.05), but feed intake was stimulated by cLEPR ECD immunization (P < 0.05). The treatment also upregulated the gene expression levels of lepR, AMP-activated protein kinase (AMPK), acetyl CoA carboxylase-2 (ACC2), and uncoupling protein 3 (UCP3) in liver, abdominal fat, and breast muscle (P < 0.05) but decreased fasn expression levels (P < 0.01). Apart from that of lepR, the expression of appetite-regulating genes, such as orexigenic genes, agouti-related peptide (AgRP) and neuropeptide Y (NPY), were upregulated (P < 0.01), whereas the anorexigenic gene proopiomelanocortin (POMC) was downregulated in the hypothalamic tissue of cLEPR-immunized pullets (P < 0.01). Blood concentrations of metabolic molecules, such as glucose, triglycerides, and very-low-density lipoprotein, were significantly decreased in cLEPR-immunized pullets but those of cholesterol, high-density lipoprotein, and low-density lipoprotein increased. These results demonstrate that antibodies to membrane proximal cLEPR ECD enhance cLEPR signal transduction, which stimulates metabolism and reduces fat deposition in chickens.