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Wang W.,Key Laboratory of Chicken Genetics and Breeding | Wang W.,Key Laboratory of Animal Genetics | Cheng M.,Key Laboratory of Chicken Genetics and Breeding | Cheng M.,Key Laboratory of Animal Genetics | And 6 more authors.
Poultry Science | Year: 2017

Gga-miR-21 is abundantly expressed in chicken pre-adipocytes, but its role is unclear. The present study investigated the role of gga-miR-21 in chicken pre-adipocyte proliferation. Cell proliferation assay and gene expression analysis of proliferating cell nuclear antigen (PCNA) showed that the gga-miR-21 mimic inhibited pre-adipocyte proliferation. In contrast, the gga-miR-21 inhibitor enhanced pre-adipocyte proliferation. The subsequent investigation identified Kruppel-like factor 5 (KLF5) mRNA as a target of ggamiR-21. The gga-miR-21 mimic inhibited KLF5 3′UTR reporter activity and decreased endogenous KLF5 expression in primary pre-adipocytes. KLF5 knockdown using RNAi had a similar effect to that of the gga-miR-21 mimic on cell proliferation. The promoting effect of the gga-miR-21 inhibitor on pre-adipocyte proliferation was partially attenuated by KLF5 knockdown. Taken together, our results demonstrated that miR-21 inhibits chicken pre-adipocyte proliferation, at least in part, by targeting KLF5. © 2016 Poultry Science Association Inc.


Fang Z.,Key Laboratory of Animal Genetics | Fang Z.,Sichuan Agricultural University | Huang F.,Key Laboratory of Animal Genetics | Luo J.,Key Laboratory of Animal Genetics | And 4 more authors.
British Journal of Nutrition | Year: 2010

The present study was conducted in a one-factorial arrangement to determine the effects of dl-2-hydroxy-4-methylthiobutyrate (dl-HMTB) on the first-pass intestinal metabolism of dietary methionine and its extra-intestinal availability. Barrows (n 6; aged 35d; weight 86kg), implanted with arterial, portal, mesenteric and gastric catheters, were fed a diet containing dl-methionine (dl-MET) or dl-HMTB once hourly and infused intramesenterically with 1% p-aminohippurate and intragastrically with [1-13C]methionine at 70mol/kg body weight per h. Arterial and portal blood samples were taken at hourly intervals until 6h of tracer infusion and pigs was then killed for collection of muscle, intestine, liver and kidney samples. The net portal appearance of methionine, expressed as the fraction of ingested directly available l-methionine, was higher (P<005) in the dl-HMTB than in the dl-MET diet, and there was no difference (P=026) in the fractional portal balance of [1-13C]methionine between the diets. [1-13C]methionine enrichment (tracer:tracee ratio; mol/100mol amino acid) in the jejunum, arterial and portal plasma, liver, kidney and muscle was also not different (P>005) between the groups. Over the 6h period after the start of feeding, the average concentration of citrulline both in the arterial and portal plasma was higher (P<005) in the dl-HMTB than in the dl-MET group, and arterial plasma ornithine and taurine concentration was also higher (P<005) in the dl-HMTB than in the dl-MET group. However, plasma urea concentration both in the arterial and portal vein was lower (P<005) in the dl-HMTB than in the dl-MET group. These results suggested that the potential difference in the first-pass use of methionine by the intestine between the dl-HMTB and dl-MET diets might affect intestinal and systemic metabolism of other amino acids, which may provide new important insights into nutritional efficiency of different methionine sources. © 2010 The Author.


Wu J.-H.,Key Laboratory of Animal Genetics | Wu J.-H.,Inner Mongolia Agricultural University | Zhang Y.-J.,Inner Mongolia Agricultural University | Zhang J.-X.,Inner Mongolia Agricultural University | And 5 more authors.
Journal of Integrative Agriculture | Year: 2012

Seasonal hair follicle activity and fibre growth in some Cashmere-bearing goats (Caprus hircus) is a cyclic process that is well characterized morphologically but understood incompletely at the molecular level. As an initial step in discovering regulators in hair-follicle activity and cycling, we used qPCR to investigate 19 genes expression in Cashmere goat side skin from 12 mon. Many of these genes may be associated with the hair follicle development-relevant genes (HFDRGs) in the literature. Here we show that Hoxc13/β-catenin gene associated with the follicle activity. In addition, Hoxc13 was found to be expressed with an drastic increase between July and November for melatonin treatments. To further investigate the role of Hoxc13 on HFDRGs, fibroblasts and keratinocytes from Cashmere goat skin were transfected with p-ECFP-Hoxc13. The result suggested that overexpression of Hoxc13 gene decreased HFDRGs with negative role for hair follicle development and increase HFDRGs with positive role for hair follicle development in vitro. These findings provide data on the Hoxc13 expression profile of normal Cashmere goat skin and Cashmere goat skin with melatonin treatment, and demonstrate hair-follicle-activity dependent regulation of Hoxc13 expression. © 2012 Chinese Academy of Agricultural Sciences.


Wang W.,Key Laboratory of Animal Genetics | Liu Y.,Nanjing Agricultural University | Wang H.,Key Laboratory of Animal Genetics | Ding X.,Key Laboratory of Animal Genetics | And 3 more authors.
Immunogenetics | Year: 2016

The interferon regulatory factor 9 (IRF9) gene is a member of the IRF family and has been shown to play functionally diverse roles in the regulation of the immune system. Previous study revealed the IRF9 gene resides within the reported quantitative trait locus (QTLs) for cytokine levels. The aims of this study were to identify genomic variants in IRF9 and to test the association between the variants and cytokine levels in pig. A synonymous single-nucleotide polymorphism (c.459A > G) was identified in exon 4 of the IRF9 gene. Association analysis in 300 piglets (Landrace, n = 68; large white, n = 158; and Songliao black, n = 74) showed that this variant was significantly associated with the level of interferon (IFN)-γ and the ratio of IFN-γ to IL-10 in serum (P < 0.05). Relative quantification of messenger RNA (mRNA) revealed that spleen had the highest expression level and individuals with genotype AA had higher expression than those with genotype AG. Transfection-based mRNA stability assay analysis further showed that the mutant allele G could reduce the RNA stability of IRF9. These findings suggest that the SNP (c.459A > G) could be a causative mutation for the association between IRF9 and the serum cytokine levels in swine. © 2015, Springer-Verlag Berlin Heidelberg.


Zhang Z.,Key Laboratory of Chicken Genetics and Breeding | Zhang Z.,Key Laboratory of Animal Genetics | Zhang Z.,Northeast Agricultural University | Wang H.,Key Laboratory of Chicken Genetics and Breeding | And 12 more authors.
Acta Biochimica et Biophysica Sinica | Year: 2013

Krü ppel-like factor 7 (Klf7) has been extensively studied in the mammalian species, but its function in avian species is unclear. The objective of this study was to reveal the function of chicken Klf7 (Gallus gallus Klf7, gKlf7) in adipogenesis. The results of real-time reverse transcription polymerase chain reaction demonstrated that the relative mRNA level of chicken Klf7 (gKlf7/gb-Actin) in the abdominal adipose tissue was significantly associated with the abdominal fat content and the age of broilers (P < 0.05), and gKlf7 was more highly expressed in preadipocytes than in mature adipocytes (P < 0.05). In addition, Oil red O staining showed that gKlf7 inhibited chicken preadipocyte differentiation, and MTT assay indicated that gKlf7 overexpression promoted preadipocyte proliferation. Additionally, luciferase assays showed that gKlf7 overexpression suppressed the chicken CCAAT/enhancerbinding protein a (C/ebpa), fatty acid synthase (Fasn), and lipoprotein lipase (Lpl) promoter activities (P < 0.05), and gKlf7 knockdown increased the chicken peroxisome proliferator-activated receptor g (Pparg), C/ebpa and fatty acid-binding protein 4 (Fabp4) promoter activities (P < 0.05). Together, our study demonstrated that chicken Klf7 inhibits preadipocyte differentiation and promotes preadipocyte proliferation. © The Author 2013.


Xie B.,Northeast Agricultural University | Wang J.,Northeast Agricultural University | Liu S.,Northeast Agricultural University | Xue B.,Northeast Agricultural University | And 5 more authors.
Cellular Reprogramming | Year: 2014

Induced pluripotent stem cells (iPSCs) and nuclear transfer (NT) are two of the primary routes to reprogram differentiated cells back to the pluripotent state. However, it is still unknown whether there is any correlation between the reprogramming efficiency of iPSCs and NT if the same donor cells are employed. In this study, six porcine embryonic fibroblast (PEF) lines from Landrace (L1, L6, L9) or Congjiang local pigs (C4, C5, C6) were used for iPSC induction and NT. Furthermore, the resultant iPSCs from four PEF lines (L1, L6, C4, and C5) were used for NT (iPSC-NT), and the expression of exogenous genes was detected in iPSC-NT embryos by real-time PCR. The results showed that the efficiency of iPSC lines established from different PEF lines were significantly different. When the same PEF lines were used as donor cells for NT, the blastocysts rates were also different among different PEF lines and positively related with iPSCs induction efficiency. When the iPSCs were used as donor cells for NT, compared with the source PEFs, the blastocysts rates were significantly decreased. Real-time PCR results indicated that exogenous genes (Oct4, c-Myc) continued to be expressed in iPSC-NT embryos. In summary, our results demonstrate that there was a positive correlation between iPSCs and NT reprogramming efficiency, although the mechanism of these two routes is different. This may provide a new method to select the appropriate donor cells for inducing iPSCs. Copyright © 2014, Mary Ann Liebert, Inc.


Liu S.J.,Key Laboratory of Animal Genetics | Liu S.J.,Heilongjiang Bayi Agricultural University | Li J.L.,Heilongjiang Bayi Agricultural University | Li H.,Key Laboratory of Animal Genetics | Li H.,Northeast Agricultural University
Animal Production Science | Year: 2014

The objective of this study was to score reactions to artificial semen collection procedures and to develop an efficient method for gander selection in two original geese breeds. One-hundred and three Zi geese and 110 Rhin geese were used. Gander reaction scores to massage and semen quality parameters were investigated in 10 Zi and 10 Rhin ganders during a complete reproductive season. The relationships between average gander massage reaction score and semen quality were evaluated. After natural mating, egg fertility comparison between Zi ganders selected by massage reaction scores and those not subjected to selection were investigated. Ganders exhibited large variations in artificial semen collection and semen characteristics between breeds. Only 30.1% (31/103) of Zi geese and 46.3% (51/110) of Rhin geese had stable positive massage reactions with semen ejaculation (P < 0.05). Average massage reaction score, ejaculate volume, spermatozoa concentration, sperm motility and semen quality factor for single ejaculate were significantly higher in Zi than in Rhin geese. There was a high correlation between average massage reaction score and semen volume, being 0.87 for Zi geese (P < 0.05) and 0.82 for Rhin (P < 0.05), and this correlation enabled pre-selection of ganders at the beginning of the breeding season. Reproductive efficiency could be improved with massage-selected ganders in natural breeding flocks. Examination of gander massage reaction scores combined with laboratory assessment of semen quality early in the breeding season is a feasible method for predicting fertilising capability of ganders. © 2014 CSIRO.


Cong Y.,Northeast Agricultural University | Ma J.,Northeast Agricultural University | Sun R.,Harbin Medical University | Wang J.,Northeast Agricultural University | And 5 more authors.
Journal of Genetics and Genomics | Year: 2013

Embryonic germ (EG) cells are cultured pluripotent stem cells derived from the primordial germ cells (PGCs) that migrate from the dorsal mesentery of the hindgut to the developing genital ridge. In this study, the morphology of the porcine genital ridge was assessed in embryos harvested on days 22-30 of pregnancy. PGCs from embryos at these stages were cultured to obtain porcine EG cell lines, and EG-like cells were derived from PGCs from embryos harvested on days 24-28 of pregnancy. The EG-like cells expressed Oct4, Sox2, Nanog, SSEA-3, SSEA-4 and alkaline phosphatase (AP). These cells were able to form embryoid bodies (EBs) in suspension culture and differentiate into cells representative of the three germ layers as verified by a-fetoprotein (AFP), α-smooth muscle actin (α-SMA), and Nestin expression. Spontaneous differentiation from the porcine EG-like cells of delayed passage invitro showed that they could differentiate into epithelial-like cells, mesenchymal-like cells and neuron-like cells. Invitro directed differentiation generated osteocytes, adipocytes and a variety of neural lineage cells, as demonstrated by alizarin red staining, oil red O staining, and immunofluorescence for neuronal class III β-tubulin (Tuj1), glial fibrillary protein (GFAP) and galactosylceramidase (GALC), respectively. These results indicate that porcine EG-like cells have the potential for multi-lineage differentiation and are useful for basic porcine stem cell research. © 2013.


Shang Z.,Key Laboratory of Chicken Genetics and Breeding | Shang Z.,Key Laboratory of Animal Genetics | Shang Z.,Northeast Agricultural University | Guo L.,Key Laboratory of Chicken Genetics and Breeding | And 14 more authors.
Bioscience Reports | Year: 2014

In addition to providing energy and constituting cell membrane, fatty acids also play an important role in adipocyte differentiation and lipid metabolism. As an important member of monounsaturated fatty acids, oleate, together with other components, is widely used to induce chicken preadipocyte differentiation. However, it is not clear whether oleate alone can induce chicken preadipocyte differentiation. In the present study, four different treatments were designed to test this question: basal medium, IDX [insulin, dexamethasone and IBMX (isobutylmethylxanthine)], oleate and IDX plus oleate. Cytoplasmic lipid droplet accumulation and mRNA expression for adipogenesis-related genes were monitored. After treatment of oleate on chicken preadipocytes, apparent lipid droplet formation and lipid accumulation were observed, accompanied by increasing expression of PPARγ (peroxisome proliferator-activated receptor-γ) and AFABP (adipocyte fatty acid-binding protein), but decreasing level of GATA2 (GATA-binding protein 2). In contrast, for cells cultured in the basal medium with or without IDX supplementation, lipid droplet barely occurred. These results suggest that exogenous oleate alone can act as an inducer of preadipocyte differentiation into adipocytes. © 2014 The Author(s).


Li Y.,Key Laboratory of Animal Genetics | Fang Z.,Key Laboratory of Animal Genetics | Dai J.,Key Laboratory of Animal Genetics | Partridge G.,Danisco | And 2 more authors.
Animal Feed Science and Technology | Year: 2010

Three experiments were conducted to assess enzymes with potential to improve the digestibility of dietary components by pigs fed diets based on extruded (EXd) or non-extruded (nEXd) corn. In Exp. 1, effects of enzyme blends (amylase, protease and xylanase) at different dose rates [high (H) vs. medium (M) vs. low (L)] on the coefficient of apparent digestibility (CAD) were investigated in vitro using a two-stage enzyme incubation method. The CAD of starch and NDF were both higher (P<0.05) in H- and M-enzyme diets than in L-enzyme and control diets. Though the CAD of DM, GE, starch and NDF did not differ between H- and M-enzyme diets, the CP CAD was higher (P<0.05) in H-enzyme than in M-enzyme diets. Exp. 2 was designed to further examine the effects of corn extrusion, the addition of phytase and its combination with H- and M-enzyme on the CAD of dietary components. In this experiment M-levels of xylanase and amylase were included in both the H- and M-enzyme blends, with protease dosage being held the same in each blend as in Exp. 1. The CAD of all dietary components evaluated was higher (P<0.05) in EXd than in nEXd corn diets. For nEXd corn diets, the combination of phytase with M- and H-enzymes resulted in a higher (P<0.05) CAD of GE, CP, NDF and starch than did phytase addition alone. In Exp. 3, both in vitro and in vivo trials were conducted to evaluate responses of corn-based diets to the extrusion process and the addition of phytase, H- and M-enzyme blends. Five cannulated pigs were fed five diets according to a 5 × 5 Latin square design. Similar to the results observed in the in vitro trial of Exp. 3, the EXd corn diet had higher (P<0.05) CAD of DM, CP, starch, NDF and GE than nEXd corn diets. M- and H-enzyme addition both resulted in increased (P<0.05) DM and CP CAD of nEXd corn diets. The CAD of amino acids of nEXd corn diets with H-enzyme addition was comparable with that of the EXd corn diet, and was higher (P<0.05) than that of the nEXd corn control diet except for Met, Thr, Trp and Cys. These results suggest that extrusion and multiple enzyme addition are effective ways to improve the nutritional value of corn-based diets for pigs. © 2010 Elsevier B.V. All rights reserved.

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