Dynamic changes in protein concentrations of keratins in crop milk and related gene expression in pigeon crops during different incubation and chick rearing stages.

1. The objective of this study was to examine the keratin composition of crop milk, the variation of epithelial thickness and keratin (K) gene expression in samples from young pigeon during incubation and chick rearing.2. Crop milk was collected from 1-, 3- and 5-day-old squab crops for keratin content analysis. Results showed that K4 accounted for the highest proportion of all detected keratins.3. In total, 42 pairs of adult pigeons were allocated to seven groups according to different stages to collect crop samples. Gene expression studies showed that the K3 gene expression was maximised at rearing Day 15 (15) and R1 in males and females, respectively. K6a gene level was the greatest at R15 in females, whereas it peaked at incubation Day 4 (I4) in males. The K12, K13, K23 and K80 gene levels were inhibited at the peak period of crop milk formation in comparison with I4. In females, K cochleal expression peaked at I10, whereas it was the greatest at R25 in males. K4 and K14 gene expression was the highest at I10 in females, while K4 and K14 were minimised at I17 and R7 in males, respectively. Gene expressions of K5, K8, K19 and K20 in males and K19 in females were maximised at R1. The K5, K20 and K75 gene levels in females peaked at R7. K75 and K8 expressions in males and females reached a maximum value at R25 and I17, respectively.4. The epithelial thickness of male and female crops reached their greatest levels at R1 and had the highest correlation with K19.5. These results emphasised the importance of keratinisation in crop milk formation, and different keratins probably play various roles during this period. The K19 was probably a marker for pigeon crop epithelium development. The sex of the parent pigeon affected keratin gene expression profiles.

In ovo feeding of creatine pyruvate modulates growth performance, energy reserves and mRNA expression levels of gluconeogenesis and glycogenesis enzymes in liver of embryos and neonatal broilers.

The effects of in ovo feeding (IOF) of creatine pyruvate (CrPyr) on the growth performance, energy reserves and mRNA expression levels of gluconeogenesis and glycogenesis enzymes in liver of late-term embryos and neonatal broilers were investigated. After candling on 16 day of incubation, a total of 960 eggs were randomly assigned to three treatments: (i) non-injected control, (ii) saline group injected with 0.6 ml of 0.75% physiological saline and (iii) Creatine pyruvate group injected with 0.6 ml of physiological saline containing 12 mg CrPyr/egg. After hatching, 120 male chicks with average body weight (BW) were randomly allocated into each treatment group for a 7-day feeding trial. The results showed that broilers subjected to CrPyr treatment had higher BW than those of the control and saline groups on 1, 3 and 7 day post-hatch, as well as the yolk sac weight on 19 day of incubation (19 E), the day of hatch and 3 day post-hatch (p < .05). Compared with the control and saline groups, IOF of CrPyr increased the plasma creatine concentration on the day of hatch, and the plasma pyruvate concentration on the day of hatch and 3 day post-hatch (p < .05). Moreover, IOF of CrPyr increased the liver pyruvate and glucose concentrations on 19 E and the day of hatch, and the liver glycogen concentration during the experiment (p < .05). Broilers in the CrPyr group showed increased mRNA expression levels of pyruvate carboxylase (PC), phosphoenolpyruvate carboxykinase (PEPCK) and glycogen synthase 2 (GYS2) on 19 E and the day of hatch (p < .05). These results indicated that IOF of CrPyr increased energy reserves in liver of embryos and neonatal broilers possibly through upregulating the mRNA expression levels of PC, PEPCK and GYS2, which could benefit the increase of BW in broilers on 7 day post-hatch.

Molecular characterization, tissue expression, and polymorphism analysis of liver-type fatty acid binding protein in Landes geese.

Liver weight is an important economic trait in the fatty goose liver industry. Liver-type fatty acid binding protein (L-FABP) is involved in the formation and metabolism of fatty acids. Thus, we hypothesized that sequence polymorphisms in L-FABP were associated with fatty liver weight in goose. We first isolated, sequenced, and characterized the goose L-FABP gene, which had not been previously reported. The goose L-FABP gene was 2490 bp and included 4 exons coding for a 126-amino acid protein. Analysis of expression levels of the goose L-FABP gene in different tissues showed that the expression level in the liver tissue was higher than in other tissues, and was significantly higher in the liver tissue of overfed geese than in control geese. Moreover, a single nucleotide polymorphism located at 774 bp in the gene was identified in a Landes goose population. To test whether this single nucleotide polymorphism was associated with fatty liver production, liver weight and the ratio of liver to carcass weights were determined for the 3 genotypes with this single nucleotide polymorphism (TT, TG, GG) in overfed Landes geese. Our data indicate that individuals with the GG genotype had higher values for the variables measured than those with the other 2 genotypes, suggesting that L-FABP can be a selection marker for the trait of fatty liver production in goose.

Lipid accumulation and oxidative stress in the crop tissues of male and female pigeons during incubation and chick-rearing periods.

The current study aimed to evaluate the changes in lipid accumulation and oxidative status in pigeon crops during different breeding stages. Forty-two pairs of adult pigeons were randomly assigned to 7 groups. Lipid droplet accumulation in pigeon crops was visualized by using oil red O staining from d 17 of incubation (I17) to d 7 of chick rearing (R7). Transmission electron microscopy analysis showed swollen mitochondria with disintegration of cristae and typical characteristics of endoplasmic reticulum stress in crop tissues at R1 compared with those at I4. During the peak of pigeon milk formation, the concentrations of reactive oxygen species, and oxidative damage markers (advanced oxidation protein products, 8-hydroxy-2 deoxyguanosine, and malondialdehyde) and the enzyme activities of total superoxide dismutase and glutathione peroxidase were all elevated significantly (P < 0.05). The protein concentration of B-cell lymphoma-2 associated X in crop tissues was significantly higher at R1, while the level of B-cell lymphoma-2 protein in males was the highest at I4 (P < 0.05). The ratio of B-cell lymphoma-2 associated X protein (Bax)/B-cell lymphoma-2 (Bcl-2) in both male and female crops peaked at R1 (P < 0.05). Gene expression of the key enzymes involved in mitochondrial and peroxisomal fatty acid ß-oxidation was investigated in crops. In males, the gene expression of carnitine palmitoyltransferase 1a peaked at R15, and that of carnitine palmitoyltransferase 2 increased significantly from R1 to R15 (P < 0.05). The mRNA abundance of long chain 3-hydroxyacyl-CoA dehydrogenase increased to the maximum value at R1 and I17 in males and females, respectively. From I17 to R7, the mRNA levels of acyl-CoA oxidase 1 and acyl-CoA oxidase 2 were decreased in pigeon crops (P < 0.05). Conclusively, lipid droplet accumulation was found in male and female pigeon crops from the end of incubation to the early stage of chick rearing. Although antioxidant defence and mitochondrial fatty acid ß-oxidation were both mobilized, oxidative stress in crop tissues still occurred during the peak of milk formation.

Effects of different stocking densities on organ development, blood biochemical indices, and antioxidative status of breeder pigeons during the rearing period.

The present experiment was conducted to investigate the effect of different stocking densities on the organ development, blood biochemical indices, and antioxidative status of breeder pigeons during the rearing period. A total of 280 (half male and half female) 40-day-old young pigeons were allocated into 4 groups, including 3 experimental groups (in compartments of the flying room): the high stocking density (HSD) (0.308 m3/bird), standard stocking density (SD) (0.616 m3/bird), and low stocking density (LSD) (1.232 m3/bird) and a caged control (0.04125 m3/bird). The results showed that the contents of corticosterone and heat shock protein 70 in males and the corticosterone content in females were higher in the control than in the other groups. The relative weight of liver, lung, and gizzard in males of the HSD group was the highest among the 4 treatments, whereas the abdominal fat index in the control group was higher than those in the other 3 treatments. Body weight and the relative weight of liver and abdominal fat in female pigeons in HSD group increased significantly. The levels of serum urea nitrogen and uric acid in pigeons of LSD group increased significantly, while the concentration of total cholesterol and the activity of alanine aminotransferase were higher in the control group. Ion (K+, Ca2+, and Na+) concentrations in female pigeon serum were also elevated in the control. The activity of antioxidant enzymes, including the total antioxidant capacity, superoxide dismutase, and glutathione peroxidase in pigeon breast muscle and liver had different degrees of inhibition when the space room was crowded. Moreover, the level of malondialdehyde in the liver of male caged pigeons was higher than that in the other treatments. In summary, rearing in cages or at a high density caused stress responses in the breeder pigeons. The stocking density of breeder pigeons during the rearing period should be ranged from 0.616 m3/bird to 1.232 m3/bird.

Identification and characterization of microRNA from chicken adipose tissue and skeletal muscle.

MicroRNA (miRNA) is small noncoding RNA that is extensively expressed in organisms. Different types play important roles in various biological processes, such as growth and development. In this study, we identified 47 miRNA in chicken adipose tissue and skeletal muscle, of which 38 were known chicken miRNA, 4 were known miRNA homologous to other species, and 5 were potentially novel miRNA. The target genes from adipose tissue and skeletal muscle were predicted. The expression assay indicated that the 10 selected miRNA were differentially expressed in different developmental stages. Both miRNA-133a and miR-1a were muscle-related, whereas miR-122 was adipose-related miRNA. Certain identified miRNA may be essential to growth and development of chicken adipose tissue and skeletal muscle. Further studies of these miRNA will help to understand their functions in growth and development of adipose tissue and skeletal muscle of poultry.

Gene expression profile in the liver tissue of geese after overfeeding.

Geese form a fatty liver after feeding on a carbohydrate-rich diet, possibly as an evolutionary adaptation to accumulate reserves for migration. To gain insight into the gene-regulation processes of hepatic steatosis in geese, we examined the profile of transcriptional expression in goose fatty liver and control liver by suppression subtractive hybridization and measured the levels of serum biochemical variables. We found 107 genes whose expression was different between the treatment and control groups. The main functions of these genes are metabolic processes, including the metabolism of carbohydrates, amino acids, and lipids. Twenty-four genes were classified using the Kyoto Encyclopaedia of Genes and Genomes pathways. Twelve genes that related to metabolic and cellular processes were confirmed by quantitative RT-PCR. A specific positive effect of feeding was observed on the expression of genes involved mainly in unsaturated fatty acids and triglyceride synthesis, and a negative effect was observed on genes involved in ß-oxidation, cholesterol metabolism, and glycolysis. The results could serve as an important reference for the development of goose breeding for fatty liver production and human liver disease research.

Probing the effects of dietary selenised glucose on the selenium concentration, quality, and antioxidant activity of eggs and production performances of laying hens.

Selenised glucose (SeGlu) is a newly invented organic selenium compound being synthesised through the selenisation reaction of glucose with NaHSe. We hypothesised that glucose could be used as a carrier for the stable low-valent organoselenium to enhance the selenium concentrations of eggs. To probe the effects of SeGlu on production performances of laying hens, egg selenium concentration, egg quality, and antioxidant indexes, 360 Hy-Line Brown laying hens were randomly assigned to three treatment groups fed with a basal diet alone or the diet supplemented with 5 or 10 mg/kg of Se from SeGlu. The results showed that SeGlu treatment not only enhanced (P < 0.001) the Se concentration in albumen and yolks, glutathione peroxidase activity, and total antioxidant capacity of eggs but also increased (P = 0.032) the Haugh unit of eggs being stored for 2 weeks, while the production performances and egg qualities of fresh eggs were not affected. Moreover, SeGlu supplementation linearly (P < 0.001) increased the scavenging ability of superoxide radicals in eggs. Briefly, SeGlu can enhance the selenium deposition and antioxidant activity of eggs, thereby meeting the nutritional requirement for Se-deficient humans.

Effect of Akt activation on apoptosis-related gene expression in the crop tissues of male and female pigeons (Columba livia).

The current study investigated whether the expression of apoptosis genes in the pigeon crops was affected by the Akt signaling pathway during crop milk formation. First, 78 pairs of adult White King pigeons were randomly assigned to 7 groups, and the expression of apoptosis-related genes and Akt signaling pathway-related proteins in the crop tissues during different breeding stages were examined. The results showed that the mRNA levels of Bak, caspase-3, caspase-6, and caspase-9 in female crops all increased and reached their highest levels at d 17 of incubation (I17). In male crops, the levels of caspase-3 and caspase-9 gene expression peaked at d 1 of chick rearing (R1). The lowest level of Bcl-2 gene expression in females was observed at I17. The expression ratios of p-Akt (Ser473)/Akt and p-Akt (Thr308)/Akt in male crops decreased to their minimum at R1, while it was observed at d 7 of chick rearing (R7) in females. Second, 36 pairs of adult pigeons were divided into 3 groups and were subjected to SC79 injections with dosages of 0, 0.02, or 0.04 mg/kg bodyweight. The SC79 injections resulted in a considerable decrease in growth performance of pigeon squabs. In male crops, the expression ratios of p-Akt (Ser473)/Akt and p-Akt (Thr308)/Akt were significantly elevated in the 0.02 mg/kg SC79 group, while in female crops, they were higher in the 0.04 mg/kg SC79 group (P < 0.05). The SC79 injection inhibited the gene expression of Bak in female crops, but enhanced the gene expression of Bcl-2 in both male and female crops. In the 0.04 mg/kg SC79 group, a 50.7 to 75.7% decrease was observed in the expression of caspase-3, caspase-6, and caspase-9 in male and female pigeon crops. Expression of the caspase-8 gene and total Akt protein in pigeon crops was not changed in different breeding stages or after SC79 injection. In conclusion, the expression of genes related to mitochondria-dependent apoptosis can be regulated by the Akt signaling pathway, which may play a potential role in pigeon milk formation.

Thiamine modulates intestinal morphological structure and microbiota under subacute ruminal acidosis induced by a high-concentrate diet in Saanen goats.

Ruminant animals are generally fed with starch-rich grain as the main energy source, and the incidence of metabolic diseases such as subacute ruminal acidosis (SARA) is high due to the intensive farming. Thiamin has been reported to alleviate SARA caused by high-concentrate diets, but the exact mechanism is not well understood. The goal of this study was to examine the role of thiamine in intestinal inflammation and microbiota caused by high-concentrate diets. The SARA model was induced by low neutral detergent fibre/starch ration to study the effects of thiamine on intestinal tissue structure and microbiota. 18 mid-lactation (148 ± 3 d in milk; milk yield = 0.71 ± 0.0300 kg/d) Saanen goats (BW = 36.5 ± 1.99 kg; body condition score = 2.73 ± 0.16, where 1 = emaciated and 6 = obese) in parities 1 or 2 were selected. The goats were randomly divided into three groups with six replicates: (1) control diet (C; concentrate:forage 30:70), (2) high-concentrate diet (H; concentrate:forage 70:30), and (3) high-concentrate diet with 200 mg of thiamine/kg of DM intake (H + T;concentrate:forage 70:30). The experimental period was lasted for 56 d. The small and large intestine, expression of inflammatory factor genes, tight junction protein genes, total antioxidant capacity, and intestinal microbiota were measured. The results showed that SARA was observed in treatment H, whereas rumen fluid pH was improved in treatment H + T. Treatment H + T also significantly repaired the intestinal tissue structure damaged by SARA, improved the total antioxidant capacity of the small intestinal mucosa, reduced mRNA expression of inflammatory factors in the small intestine tissue, and increased the mRNA expression of tight junction genes in small intestine tissue. The high-concentrate diet reduced the diversity of intestinal microbiota. When thiamine is added to the high-concentrate diet, the relative abundance of intestinal Firmicutes and beneficial bacteria represented by Lactobacilli were upregulated, and the relative abundance of Proteus, a marker of intestinal dysbacteriosis, returned to normal. In conclusion, thiamine supplementation could alleviate the damage to the intestinal tissue structure and microbial environment caused by SARA condition in dairy goats fed a high-concentrate diet.

Effects of incubation and chick rearing on intestinal morphology, digestive enzyme activities, and mRNA expression of nutrient transporter genes in the pigeon (Columba livia) under artificial farming conditions.

The present study investigated the changes in morphology, enzyme activities in the pancreas and mucosa, and nutrient transporter gene expression in the duodenum and jejunum in male and female pigeons during the incubation and chick-rearing periods. Forty-two pairs of White King pigeons with 2 fertile eggs per pair were randomly divided into 7 groups by different breeding stages. The crypt depth of the duodenum and jejunum reached the peak at day 1 (R1) and day 7 (R7) of chick rearing, respectively. The jejunum surface area increased to a maximum value at R1. Amylase activity in the pancreas decreased to the lowest value at R1, whereas trypsin and lipase activities peaked at 17 D of incubation (I17) and R7, respectively. In male pigeons, mucosal Na+-K+-ATPase activity in the duodenum and jejunum was the highest at R15 and it was at I17 in female pigeons. Jejunum sucrose activity in female pigeons was higher at I4 than that at I17 (P < 0.05). The gene expression of FAT/CD36 and I-FABP in the duodenum gradually increased and then declined in the late chick-rearing period. SGLT1 in the jejunum decreased to a lower level at I17 and R25 in male pigeons (P < 0.05). GLUT2 expression in female duodenum and male jejunum decreased to a lower value at I17 compared with that at R15 (P < 0.05). In the late of incubation (from I10 to I17), expression of duodenum CAT1, B0AT1, and PepT1 and jejunum CAT1, ASCT1, and PepT1 in female pigeons was significantly reduced (P < 0.05), whereas opposite results were found in male jejunum CAT1 and duodenum ASCT1. In conclusion, variations of intestinal morphology, activities of pancreatic and mucosal enzymes, and gene expression of nutrient transporters during incubation and chick-rearing periods, underlying potential changes of digestive and absorptive function and intestinal adaptation with sexual effects, may represent a complicated response to stimuli of different breeding stages.

The profiling of amino acids in crop milk and plasma and mRNA abundance of amino acid transporters and enzymes related to amino acid synthesis in the crop tissue of male and female pigeons during incubation and chick-rearing periods.

The present study was carried out to investigate the changes in amino acid (AA) contents of crop milk and plasma and mRNA abundance of AA transporters and AA synthesis-related enzymes in the crop tissue of male and female pigeons during incubation and chick-rearing periods. Forty-two pairs of adult White King pigeons with 2 fertile eggs per pair were randomly divided into 7 groups by different breeding stages. The AA content of crop milk decreased from day 1 (R1) to day 25 (R25) of chick rearing (P < 0.05). In both male and female adult pigeons, the contents of Thr, Leu, Val, His, Asp, and Pro in plasma increased to maximum levels on R25. Parental sex effect and interaction between stage and sex were observed in the AA contents of pigeon plasma (P < 0.05). For AA transporters, the mRNA abundances of SNAT2, ASCT1, LAT1, and y+LAT2 in the male crops reached the highest value on day 17 of incubation (I17), and the peak mRNA levels of PAT-1, xCT, b0,+AT, and CAT1 were found on R7 (P < 0.05). In females, the abundances of ASCT1, B0AT1, asc-1, and CAT1 mRNA peaked on R1, whereas the maximum levels of LAT1, PAT-1, b0,+AT, and y+LAT2 were observed on R7. For enzymes involved in AA synthesis, the highest gene expressions of glutamate dehydrogenase 1, acetolactate synthase in both parent pigeons, and L-threonine 3-dehydrogenase in female pigeon crops were attained on I17. The expressions of ornithine-δ-aminotransferase, glutamic-oxal(o)acetic transaminase 1, glutamic-oxal(o)acetic transaminase 2, asparagine synthetase, serine hydroxymethyltransferase 2, and glutamic-pyruvic transaminase 2 in both sexes and argininosuccinate lyase and L-threonine 3-dehydrogenase in males were the highest on R1. In conclusion, AA used for pigeon crop milk formation may originate from plasma and intracellular synthesis. The genes involved in AA transport and synthesis varied significantly with sexual effects, indicating that other factors should be considered in future explorations of the mechanism of protein formation in crop milk.

Prolactin induces lipid synthesis of organ-cultured pigeon crops.

The objective of this research was to examine the effects of prolactin (PRL) on the lipid synthesis of organ-cultured pigeon crops in vitro. In experiment 1, the histology, activities of enzymes, and expression of genes involved in metabolism and apoptosis of organ-cultured pigeon crops were analyzed over a 7-d culture period. The results showed that cultured crops maintained their structural integrity for up to 3 d in vitro. Beyond 3 d, caspase-3 activity and Bak1 gene expression increased with day of culture, whereas the activities of succinate dehydrogenase, Na+-K+-ATPase, Ca2+-Mg2+-ATPase, total ATPase, and gene expression of Bcl-2 and CK-19 diminished (P < 0.05). In experiment 2, the crops were cultured for 24, 36, and 48 h in medium containing 0, 25, or 50 ng/mL PRL, respectively, and the accumulation of lipid droplets, lipid content, and expression of fatty acid transportation- and lipogenesis-related genes were analyzed. The results showed that the crops with PRL supplements showed higher amounts of lipid droplets than those of the controls, and the droplets were mainly located in the basal nutritive layer in response to PRL. The efficacy of inducing lipid accumulation increased as the concentration of PRL increased. Crops with 50 ng/mL PRL incubated for 36 h displayed the maximal lipid content. Increasing the concentration of PRL from 0 to 50 ng/mL resulted in a dose-dependent increase in the expression of acetyl-CoA carboxylase, fatty acid synthase, fatty acid translocase, fatty acid binding protein 5, acyl-CoA binding protein, and peroxisome proliferator-activated receptor γ genes after incubation for 36 h (P < 0.05). Therefore, our results indicated that the organ-cultured pigeon crops maintained good viability for up to 3 d in vitro. Furthermore, PRL induced the lipid synthesis of organ-cultured pigeon crops in a dose- and time-dependent manner, which was related to the increased expression of genes involved in fatty acid transportation and lipogenesis.

Fatty acids modulate the expression of pyruvate kinase and arachidonate-lipoxygenase through PPARγ/CYP2C45 pathway: a link to goose fatty liver.

Cytochrome P-450 2C45 (CYP2C45) is the most highly expressed cytochrome P-450 isoform in chicken liver, and may play an important role in avian liver biology. However, information regarding the function of CYP2C45 in fatty liver is generally limited. The aim of this study was to investigate the role of CYP2C45 during the development of goose fatty liver. Our result indicated that the transcription of CYP2C45, together with PK and ALOX5, was increased in goose liver upon overfeeding for 19 D (P < 0.05). In goose primary hepatocytes, CYP2C45 RNA expression was also upgraded by the treatment with various chemicals like insulin, the fatty acids, and PPAR agonists (P < 0.05). We also found that both CYP2C45 overexpression and troglitazone treatment could increase the expression of pyruvate kinase (PK) and arachidonate 5-lipoxygenase (ALOX5), and furthermore, showed that the up-regulation of PK and ALOX5 induced by troglitazone could be suppressed by small interfering RNAs targeting CYP2C45 (P < 0.05). These findings suggest that fatty acids treatment and the overfeeding can induce the up-regulation of CYP2C45 expression possibly via PPARγ and that the induction of PK and ALOX5 in goose fatty liver is at least partially attributed to fatty acid-induced expression of CYP2C45. Thus, our data provides an insight into the mechanism by which glycolysis and arachidonic acid metabolism are modulated in goose fatty liver.

Changes in hormone profiles, growth factors, and mRNA expression of the related receptors in crop tissue, relative organ weight, and serum biochemical parameters in the domestic pigeon (Columba livia) during incubation and chick-rearing periods under artificial farming conditions.

The present study was conducted to determine the changes in concentrations of hormones and growth factors and their related receptor gene expressions in crop tissue, relative organ weight, and serum biochemical parameters in male and female pigeons during incubation and chick-rearing periods under artificial farming conditions. Seventy-eight pairs of 60-week-old White King pigeons with 2 fertile eggs per pair were randomly divided into 13 groups by different breeding stages. Serum prolactin and insulin-like growth factor-1 (IGF-1) concentrations in crop tissue homogenates were the highest in both male and female pigeons at 1 d of chick-rearing (R1), while epidermal growth factor (EGF) in female pigeons peaked at d 17 of incubation (I17) (P < 0.05). mRNA expression of the prolactin and EGF receptors in the crop tissue increased at the end of incubation and the early chick-rearing stage in both sexes. However, estrogen, progesterone, and growth hormone receptor expression each decreased during the early chick-rearing stage (P < 0.05). In male pigeons, IGF-1 receptor gene expression reached its peak at R7, while in female pigeons, it increased at the end of incubation. The relative weight of breast and abdominal fat in both sexes and thighs in the males was lowest at R7, and then gradually increased to the incubation period level. Serum total protein, albumin, and globulin concentrations increased to the highest levels at I17 (P < 0.05). Total cholesterol, triglyceride, and low-density lipoprotein reached their highest values at I17 in male pigeons and R25 in female pigeons (P < 0.05). In conclusion, hormones, growth factors, and their receptors potentially underlie pigeon crop tissue development. Changes in organs and serum biochemical profiles suggested their different breeding-cycle patterns with sexual effects.

Transcriptome analysis of hepatic gene expression and DNA methylation in methionine- and betaine-supplemented geese (Anser cygnoides domesticus).

Dietary methionine (Met) restriction produces a coordinated series of transcriptional responses in the liver that limits growth performance and amino acid metabolism. Methyl donor supplementation with betaine (Bet) may protect against this disturbance and affect the molecular basis of gene regulation. However, a lack of genetic information remains an obstacle to understand the mechanisms underlying the relationship between Met and Bet supplementation and its effects on genetic mechanisms. The goal of this study was to identify the effects of dietary supplementation of Met and Bet on growth performance, transcriptomic gene expression, and epigenetic mechanisms in geese on a Met-deficient diet. One hundred and fifty 21-day-old healthy male Yangzhou geese of similar body weight were randomly distributed into 3 groups with 5 replicates per treatment and 10 geese per replicate: Met-deficient diet (Control), Control+1.2 g/kg of Met (Met), and Control+0.6 g/kg of Bet (Bet). All geese had free access to the diet and water throughout rearing. Our results indicated that supplementation of 1.2 g/kg of Met in Met-deficient feed increased growth performance and plasma homocysteine (HCY) levels, indicating increased transsulfuration flux in the liver. Supplementation of 0.6 g/kg Bet had no apparent sparing effect on Met needs for growth performance in growing geese. The expression of many genes critical for Met metabolism is increased in Met supplementation group. In the Bet-supplemented group, genes involved in energy production and conversion were up-regulated. Dietary supplementation with Bet and Met also altered DNA methylation. We observed changes in the methylation of the LOC106032502 promoter and corresponding changes in mRNA expression. In conclusion, Met and Bet supplementation in geese affects the transcriptional regulatory network and alters the hepatic DNA methylation of LOC106032502.