Comparative transcriptome analysis reveals genes related to the yolk ratio of duck eggs.

Yolk ratio is an important production index in the salted duck egg industry. Yolk constituents are deposited during development of follicles. However, the molecular mechanism responsible for different yolk ratios in laying ducks remains elusive. In this study, Shaoxing ducks laying eggs with different yolk ratios were chosen for an analysis of liver and ovary transcriptome information. Twelve libraries were constructed and generated an average of 58.5 million clean reads per library, of which 69% of clean reads from liver and 65% of clean reads from ovary were mapped to a reference genome. Between cross-phenotype groups, a total of 250 and 230 differently expressed genes (DEGs) were identified in liver and ovary respectively, of which 101 and 50 DEGs respectively were characterized. Several DEGs were detected, among which HMGCS1, HMGCR, FDFT1, (DHCR7), (STARD4), CYP46A1 and LPIN3 are involved in cholesterol metabolism-related pathways; KIAA0319, STARD4, AP1S3, SH3GL2 and CAV2 are involved in vesicular transport in the liver; and ELOVL2 and PSD2 are involved in fatty acid elongation and endocytosis in the ovary. High yolk-ratio ducks had higher activity for cholesterol synthesis and molecular trafficking. The identification of candidate genes greatly advances the understanding of the genetic basis of the formation of different yolk ratios.

Cloning, expression and bioinformatics analysis of a putative pigeon melanoma differentiation-associated gene 5.

1. Melanoma differentiation-associated gene 5 (MDA5) is a critical member of cytosolic pattern recognition receptors (PRRs) that recognise viral RNA and mediate type I interferon secretion in host cells. 2. The objective of the present study was to identify and characterise the structure and expression of pigeon MDA5. 3. The full-length MDA5 cDNA was cloned from pigeon spleen using RT-PCR and RACE. The distribution and expression level of pigeon MDA5 in different tissues were determined by QRT-PCR. 4. The results showed that the full-length pigeon MDA5 cDNA had 3858 nucleotides (containing a 210-bp 5'-UTR, a 3030-bp open reading frame and a 618-bp 3'-UTR) encoding a polypeptide of 1009 amino acids. The deduced amino acid sequence contained six conserved structural domains typical of RIG-I-like receptor (RLR), including two tandem arranged N-terminal caspase activation and recruitment domains (CARDs), a DEAH/DEAD box helicase domain (DExDc), a helicase superfamily c-terminal domain (HELICc), a type III restriction enzyme (ResIII) and a C-terminal regulatory domain (RD). 5. The pigeon MDA5 showed 84.8%, 87.3%, 87.9% and 87.2% amino acid sequence identities with previously described homologues from chicken, duck, goose and Muscovy ducks, respectively, and phylogenetic analysis revealed a close relationship among these MDA5. 6. Pigeon MDA5 transcript was ubiquitously expressed in all seven tissues tested in healthy pigeons and showed a high level in the thymus gland and kidney. 7. These findings lay the foundation for further research on the function and mechanism of MDA5 in innate immune responses related to vaccinations and infectious diseases in the pigeon.

RNA interference of leptin receptor in chicken adipocytes.

In this study, chicken adipocytes were cultured to evaluate RNA interference by the leptin receptor gene. A small interfering RNA of the leptin receptor gene was synthesized, with a suppression rate of 60% being generated (P < 0.01). After the knockdown of the leptin receptor, the expression levels of certain genes decreased significantly; specifically, peroxisome proliferator-activated receptor γ, fatty acid synthase, adipose triglyceride lipase, and lipoprotein lipase. In addition, a significant increase in the expression of the adiponectin gene was documented. These results demonstrate that the leptin receptor gene might contribute to lipid metabolism by influencing the expressions of the peroxisome proliferator-activated receptor γ, fatty acid synthase, adipose triglyceride lipase, lipoprotein lipase, and adiponectin genes.

Molecular cloning, expression, and regulation of estrogen receptors in pigeon oviduct epithelial cells.

Estrogen regulates reproductive behavior and drives the proliferation and differentiation of several cell types. These physiological functions of estrogen are mediated by estrogen receptors (ERs), and each ER isoform plays a distinct role. To clarify the molecular mechanism of estrogen action and to evaluate the effect of ERs on the secretion of ovalbumin (OVA) in pigeon oviduct epithelial cells (POECs), we determined the complete coding sequences encoding ER alpha (ERα) and ER beta (ERß) in pigeons. The abundance of pigeon ERα and ERß mRNA was detected using quantitative polymerase chain reaction. These results revealed that pigeon ERα is highly expressed in the oviduct, while pigeon ERb is highly expressed in the ovary and kidney. We hypothesize that ERα mRNA predominates over that of ERß in the oviduct. The expression of ERα can be down-regulated by 17ß-estradiol, and the knockdown of ERα promoted OVA mRNA expression in cultured POECs, indicating that ERα may play an important role in OVA secretion.

Molecular cloning, expression, and regulation of the ovalbumin gene in pigeon oviduct epithelial cells.

The full-length pigeon ovalbumin (OVA) gene cDNA was cloned and sequenced by reverse transcription-polymerase chain reaction (RT-PCR) and rapid-amplification of cDNA ends. A 386-amino acid protein was predicted for the obtained sequence, which had 67% identity with the chicken protein. Similar to chicken OVA, the pigeon OVA gene is a non-inhibitory serine protease inhibitor. Quantitative PCR analysis revealed that pigeon OVA mRNA was highly expressed in the oviduct, and trace amounts were detected in other tissues. During the reproductive cycle, pigeon oviduct OVA mRNA expression reached its peak during the egg-laying stage, decreased with brooding, and then increased again during the squab-feeding period. Moreover, the relative OVA expression level in pigeon oviduct epithelial cells could be upregulated by a constant concentration of steroid hormones.

Construction and analysis of a subtractive cDNA library of early embryonic development in duck.

Several studies have documented the process of early embryonic development in poultry; however, the molecular mechanisms underlying its developmental regulation are poorly understood, particularly in ducks. In this study, we analyzed differential gene expression of embryos 6 and 25 h following oviposition to determine which genes regulate the early developmental stage in ducks. Among 216 randomly selected clones, 39 protein-encoding cDNAs that function in metabolism, transcription, transportation, proliferation/apoptosis, cell cycle, cell adhesion, and methylation were identified. Additionally, the full-length cDNA of the Nanog gene, encoding a 302-amino acid protein, was obtained. Quantitative real-time polymerase chain reaction analyses were performed to detect expression levels of the selected genes during early and late embryonic stages, which revealed that these genes are expressed in a particular spatial and temporal pattern. These results indicate that these genes may play pivotal roles in the process of area pellucida formation through a complex and precise regulatory network during development in duck embryos.

Expression pattern of adipocyte fatty acid-binding protein gene in different tissues and its regulation of genes related to adipocyte differentiation in duck.

Abundance of duck adipocyte fatty acid-binding protein (A-FABP) mRNA was detected in this study by quantitative real-time PCR. The results showed that duck A-FABP gene was expressed in muscular tissues and many organs, except pancreas, lung, and kidney, and highly expressed in adipose tissues, especially in sebum. The expression of A-FABP and adipocyte differentiation-related genes was upregulated by oleic acid, and the A-FABP knockdown suppressed the oleic acid-stimulated expression of these genes in the cultured duck adipocytes, indicating A-FABP might play an important role in duck adipocyte differentiation.

Effects of linoleic acid and eicosapentaenoic acid on cell proliferation and lipid-metabolism gene expression in primary duck hepatocytes.

Several studies have investigated that linoleic acid (LA) and eicosapentaenoic acid (EPA) affect cell proliferation and lipid catabolic gene expression in mammals. To determine if LA and EPA increase duck cell proliferation and lipid catabolic gene expression, the authors exposed duck primary hepatocyte cultures to LA or EPA. The results showed that both LA and EPA increased cell proliferation in a dose-dependent manner (100 µM). The effect on specific cell-cycle phases was also studied; LA and EPA (100 µM) deceased the proportion of cells in the G0/G1 phase from 83 to 80.8 and 80.3%, respectively, concomitant with an increase in the proportion of S-phase cells (11.5 and 10.5 vs. 8%, respectively). The expression of PPAR-α and PPAR-α target genes, such as acyl-CoA oxidase (ACOX), lipoprotein lipase (LPL), liver fatty acid-binding protein (L-FABP), was examined by quantitative real-time PCR. The results showed that the expression of the PPAR-α, ACOX, and LPL genes increased significantly following LA and EPA exposure, but that the expression of L-FABP remained unchanged. This study provides the first characterization of LA- and EPA-induced cell proliferation and PPAR-α and PPAR-α target gene transcriptional responses in duck primary hepatocyte cultures.

Effects of perilla extract on productive performance, serum values and hepatic expression of lipid-related genes in Shaoxing ducks.

1. The aim of this study was to identify the effect of perilla extract, a source of polyunsaturated fatty acids, on lipid metabolism and expression of lipid-related genes in livers of Shaoxing ducks. 2. Two hundred and forty 28-week-old laying ducks received a commercial diet with perilla extract added at 0 (control) or 200 mg/kg of feed. 3. Ducks fed on a diet with perilla extract had increased laying rates compared with control ducks. 4. Serum concentrations of triglycerides were reduced by perilla extract, while high-density lipoprotein cholesterol and total serum cholesterol increased. 5. The expression of genes involved in hepatic lipogenesis, sterol regulatory element-binding protein-1, acetyl CoA carboxylase, stearoyl CoA desaturase, fatty acid synthase, apolipoprotein B, and apolipoprotein very low density lipoprotein, were decreased in the perilla group. 6. The mRNA expression of peroxisome proliferators-activated receptor alpha and acyl-coenzyme A oxidase was enhanced following treatment with perilla extract, and a similar tendency was observed in the expression of liver fatty acid-binding protein. 7. The results show that a diet with 200 mg/kg perilla extract regulated fat metabolism of Shaoxing ducks by improving egg laying, altering serum lipid profiles, stimulating lipid catabolic gene expression and inhibiting lipogenic gene expression in the liver.

Effects of parental dietary linoleic acid on growth performance, antioxidant capacity, and lipid metabolism in domestic pigeons (Columba livia).

The objective of this study was to evaluate the effects of dietary linoleic acid (LA) on growth performance, antioxidant capacity, and lipid metabolism in pigeon squabs by supplementing LA in their parental diets. A completely randomized design that consisted of a control group, 1% dietary LA addition group (LA1%), 2% dietary LA addition group (LA2%), and 4% dietary LA addition group (LA4%) was used. Six squabs from each treatment were randomly sampled at the day of hatch and days 7, 14, and 21 after hatch. The results showed that parental dietary LA had no significant influence (P > 0.05) on body weight (BW) gain or relative organ weights (% of BW) in squabs. The activities of superoxide dismutase, catalase, and glutathione peroxidase in the LA1% were significantly increased (P < 0.05) compared with those in the control group. The malondialdehyde content in the LA1% was significantly lower (P < 0.05) than that in the control group. The levels of serum triglyceride in the LA1% and LA2% were significantly decreased (P < 0.05) compared with those in the control group, whereas the serum high-density lipoprotein cholesterol level in the LA1% and LA2% and the free fatty acid level in the LA4% were significantly higher (P < 0.05) than those of the control group. The activities of lipoprotein lipase, hepatic lipase, and hormone-sensitive lipase in the LA1% were significantly higher (P < 0.05) than those in the control group. The 3-hydroxy-3-methylglutaryl coenzyme A reductase activity in the LA1% and the hormone-sensitive lipase activity in the LA4% were significantly decreased (P < 0.05) compared with those in the control group. The mRNA expression of carnitine palmitoyltransferase 1, acyl-CoA 1, and peroxisome proliferator-activated receptor α was significantly upregulated (P < 0.05) in the LA1% compared with that in the control group. The Oil Red O staining area in the LA1% and LA2% was significantly reduced compared with that in the control group. The results indicated that although supplemental LA had negligible effects on growth and development in pigeon squabs, parental dietary LA at a concentration of 1% could have beneficial effects on maintaining squabs healthy as reflected by improved antioxidant capacity and lipid metabolism.

Effect of daidzein on egg-laying performance in Shaoxing duck breeders during different stages of the egg production cycle.

The effect of a phyto-oestrogen, daidzein, on the laying performance of Shaoxing female ducks was examined in three experiments performed at different stages of the egg production cycle. Egg-laying rate, egg weight, egg composition, feed conversion ratio, hatchability characteristics of eggs and body weight, ovary and oviduct weight, as well as changes in serum concentrations of T3, T4 and E2 were recorded as response criteria. In the first experiment, 68 young ducks, 100 d of age, were given a basal diet (maize-soybean meal) with or without 3 mg of daidzein/kg diet for 42 d. Daidzein did not affect the onset of lay but apparently decreased egg-laying rate and mean egg weight as well as the feed conversion ratio. In the second experiment, 240 breeding ducks, 402 d of age, were allotted at random to three groups and given the basal diet containing daidzein at 0 (control), 3 (Da1) and 5mg/kg (Da2) for 35d. Egg-laying rate, mean egg weight and feed conversion ratio increased in both Da1 and Da2 groups. However, an adverse effect of daidzein on fertility and hatchability was observed. In the third experiment, 320 breeding ducks, 415 d of age, were fed on the basal diet with or without 5mg of daidzein/kg diet for 63 d. Egg-laying rate increased by 7.7%, average egg weight tended to increase, whereas yolk/albumen ratio decreased. Daidzein-treated ducks had higher body weight and oviduct weight compared with their controls. Elevated plasma T4 and E2 concentrations accompanied these phenotypic changes, but serum T3 was not affected. It is suggested that daidzein exerts divergent effects on the egg-laying performance of Shaoxing ducks under different physiological conditions and this action is dose-dependent. The changes in circulating E2 imply possible participation of endogenous oestrogen in the mechanism of daidzein action.