Effect of maternal rearing on lipid metabolism-related gene expression in offspring broilers during embryonic development

1. The aim of this study was to investigate the effect of maternal rearing on lipid metabolism and lipid metabolism-related gene expression in offspring broilers during embryonic development. 2. One hundred laying Sanhuang breeders were divided into two groups, and either floor-reared or cage-reared on the same diet. Liver and serum samples were extracted on days 14 and 19 of embryonic development and at hatching. The lipid metabolism related gene expressions of acetyl CoA carboxylase (ACC), fatty acid synthase (FAS), malic enzyme (ME), apolipoprotein B100(apoB100), sterol regulating element binding protein (SREBP-1c), carnitine palmitoyltransferase (CPT-1) and peroxisome proliferators-activated receptor (PPARα) genes were determined using real time RT-PCR. 3. The results showed that embryonic weight, liver weight, serum and hepatic total cholesterol (TC) concentration and serum triglyceride (TG) content were not significantly different between the cage-reared group and the floor-reared group during embryonic development. However, embryonic weight, liver weight, serum and hepatic TC concentration and serum TG content in the cage-reared group were significantly higher than in the floor-reared group at hatching. 4. Hepatic ACC, FAS, SREBP-1c, ME and apoB genes expression were not significantly different between the cage-reared and the floor-reared groups during E9 and E14 development. Hepatic ME gene expression in the cage-reared group was higher than in the floor-reared group during E19 development. However, hepatic FAS, SREBP-1c, CPT-1 and PPARα gene expressions in the cage-reared group was higher than in the floor-reared group. 5. A change in the maternal regime could regulate lipid metabolism in offspring broilers during embryonic development, and especially at hatching.     

胚胎及新生期蛋鸡和肉鸡垂体生长激素基因表达的比较研究

本文采用Northern杂交分析法研究了14胚龄、18胚龄、1日龄、5日龄及10日龄肉鸡和蛋鸡垂体生长激素的基因表达，同时采用放射免疫分析法（RIA）测定了垂体内生长激素的含量。结果表明：14胚龄时在肉鸡和蛋鸡垂体总RNA中均未出生长激素mRNA(GH mRNA)。从18胚龄开始可检测出一条0.8kb的GHmRNA，并且垂体GHmRNA水平的发育性变化在品种间呈现不同的规律：蛋鸡从18胚龄到10日龄垂体GHmRNA水平不断升高，日龄间差异显著（P＜0.05）。肉鸡从18胚龄到出壳1日龄，GHmRNA水平有较大幅度的升高，但从1日龄至10日龄维持在1日龄时的水平。18胚龄肉鸡垂体GH mRNA水平显著高于蛋鸡（P＜0.05），并在1日龄和5日龄均维持在较高水平，与生长速度呈正相关；而10日龄时垂体HGmRNA水平的品种差异发生逆转，蛋鸡GHmRNA水平反而高于肉鸡。垂体HG含量的发育性变化趋势与垂体HGmRNA水平相一致，10日龄时蛋鸡垂体GH含量显著高于肉鸡（P＜0.05），与生长速度呈相反的趋。     

Total lipid and triacylglycerol contents in the liver of broiler and layer chickens at embryonic stages and hatching

To compare the hepatic function of broiler and layer chickens (Gallus gallus domesticus) at various embryonic stages and hatching, the total lipid and triacylglycerol (TG) content were determined. The chicken embryos accumulated a large amount of lipids in the liver and the total lipid content gradually increased toward hatching, though no significant difference was observed between broilers and layers. The TG contents in the liver increased considerably with developmental stage. At embryonic day 14 the TG content in the liver was similar between broilers and layers; thereafter, it was 1.3 and 2.2 times higher in broilers than in layers at embryonic day 18 and for newly hatched chicks, respectively. Chick embryos accumulate an excessive amount of cholesterol ester in the liver, but cholesterol ester is replaced by TG after hatching. The results of hepatic TG contents in the present study suggest that the development of the hepatic function between broilers and layers may already differ at embryonic stages.     

Based serum metabolomics analysis reveals simultaneous interconnecting changes during chicken embryonic development

Metabolic disorder is a major health problem and is associated with a number of metabolic diseases. Due to native hyperglycaemia and resistance to exogenous insulin, chickens as a model had used in the studies of adipose tissue biology, metabolism and obesity. But no detailed information is available about the comprehensive changes of serum metabolites at different stages of chicken embryonic development. This study employed LC/MS‐QTOF to determine the changes of major functional metabolites at incubation day 14 (E14d), 19 (E19d) and hatching day 1 (H1d), and the associated pathways of differential metabolites during chicken embryonic development were analysed using Metabolite Set Enrichment Analysis method. Results showed that 39 metabolites were significantly changed from E14d to E19d and 68 metabolites were significantly altered from E19d to H1d in chicken embryos. Protein synthesis was promoted by increasing the concentrations of L‐glutamine and threonine, and gonadal development was promoted through increasing oestrone content from E14d to E19d in chicken embryos, which indicated that serum glutamine, threonine and oestrone contents may be considered as the candidate indicators for assessment of early embryonic development. 2‐oxoglutaric acid mainly contributed to enhancing the citric cycle, and it plays an important role in improving the growth of chicken embryos at the late development; the decreasing of L‐glutamine, L‐isoleucine and L‐leucine contents from E19d to H1d in chicken embryonic development implied their possible functions as the feed additive during early posthatch period of broiler chickens to satisfy the growth. These results provided insights into understand the roles of serum metabolites at different developmental stages of chicken embryos, it also provides available information for chicken as a model to study metabolic disease or human obesity.     

Differential expression of genes implicated in liver lipid metabolism in broiler chickens differing in weight

ABSTRACT

1. Lipid parameters and expression of ACACA, APOA1, CPT1A, FASN, FOXO1, LIPG, PPARα and SIRT1 genes involved in lipid metabolism were investigated in two groups of high (HW) and low (LW) weight broilers from the same strain.

2. Blood cholesterol and liver triglyceride levels were significantly increased in HW chickens compared to LW broilers, while other parameters, i.e. blood triglyceride, blood HDL/LDL, liver cholesterol and total liver fat showed no significant changes in either group.

3. The relative expression of ACACA, APOA1 and CPT1A genes was significantly lower in the liver tissues of HW broilers than in the LW group. The mRNA levels of these three genes showed a significant negative correlation with abdominal fat deposition and live weight of broilers. However, relative expression of FASN, FOXO1, LIPG, PPARα and SIRT1 hepatic genes did not differ among broilers.

4. It was concluded that, of eight hepatic genes implicated in lipid metabolism, only the expression of three (ACACA, APOA1 and CPT1A) were significant for fat and leanness within the same strain of chicken. Since reducing body fat is a major goal in the broiler industry, these data can provide fresh insight into the molecular processes underlying the regulation of fat deposition in broilers.     

散养黄羽肉鸡CAPN1、H-FABP基因表达的发育性变化及其肌内脂肪含量的研究

为探讨在散养条件下黄羽肉鸡CAPN1、H-FABP基因与其生长发育和肌内脂肪(IMF)含量的关系,试验以130羽3周龄体重接近,健康商品代黄羽肉鸡母鸡为试验材料,随机分为笼养组(对照组)和散养组(试验组),采集在不同周龄(3、4、6、8、10、13 w)的胸肌、腿肌和腹脂组织样品共360份,采用实时荧光定量PCR法对不同生长阶段组织中CAPN1和H-FABP基因mRNA表达量进行检测,同时采用索氏抽提法测定胸肌和腿肌的IMF含量。结果表明:黄羽肉鸡的CAPN1、H-FABP基因在胸肌、腿肌和腹脂中均有不同程度的表达。随着周龄的增加,散养肉鸡CAPN1基因表达的整体变化趋势为增加—减少—增加—减少,且3～6 w其在胸肌和腹脂中的表达量均表现为笼养显著高于散养(P<0.05)。H-FABP基因的整体表达趋势随着周龄的增加而降低,在胸肌和腿肌中变化趋势相同,且均表现为散养显著高于笼养(P<0.05),在腹脂中表现为笼养高于散养(P>0.05)。与散养鸡相比,笼养鸡生长速度较快,腹脂率、IMF含量较高。本试验结果可为散养黄羽肉鸡CAPN1和H-FABP基因分子选育提供一定参考。     

Effects of epigallocatechin gallate on lipid metabolism and its underlying molecular mechanism in broiler chickens

The objective of this study was to investigate the effects of epigallocatechin gallate (EGCG) on fat metabolism and to establish the molecular mechanism of these effects in broilers. Seventy‐two 28‐day‐old male Ross 308 broiler chickens were divided into three groups with different levels of EGCG supplementation for 4 weeks: normal control (NC) group, L‐EGCG (a low‐level supplement of EGCG, 40 mg/kg body weight daily) and H‐EGCG (a high‐level supplement of EGCG, 80 mg/kg body weight daily). After 4 weeks of oral administration, EGCG significantly reduced the level of abdominal fat deposition in broilers. The serum triglycerides and low‐density lipoprotein cholesterol of chickens in H‐EGCG group were also significantly decreased compared with the NC group, and the high‐density lipoprotein cholesterol was notably increased at the same time. Moreover, the vital role of the liver and abdominal adipose tissue in lipid metabolism of poultry animals was examined through gene expression and enzyme activities related to fat anabolism and catabolism in these organs. Our data show that EGCG supplementation for 2 weeks significantly downregulated the expression of fatty acid synthesis and fat deposition‐related genes, and upregulated the expression of genes involved in fatty acid β‐oxidation and lipolysis genes. Simultaneously, the activities of hepatic fatty acid synthesis enzymes (fatty acid synthase and acetyl CoA carboxylase) were significantly decreased, and the activity of carnitine palmitoyl transferase‐1 was notably elevated. The results suggest that EGCG could alleviate fat deposition in broilers through inhibiting fat anabolism and stimulating lipid catabolism in broilers.     

日粮蛋白质水平对乌金猪肝脏组织脂类代谢相关基因表达的影响

为探讨日粮蛋白质水平对乌金猪肝脏组织脂类代谢相关基因表达的影响,试验选取体重约15 kg的乌金猪54头,随机分为3组,每组18头,为低、中和高蛋白质组,其在15～30、30～60及60～100 kg生长阶段的蛋白质水平分别为14%、12%、10%,16%、14%、12%和18%、16%、14%。猪在体重为30、60和100 kg时屠宰,取其肝脏组织,Real-time PCR定量检测脂类代谢相关基因的表达水平。结果显示,在30、60和100 kg时,乌金猪肝脏组织中ACC、FAS、SREBP-1C基因的表达水平均随着蛋白质水平的升高而降低;CPT-Ⅰ和PPARα基因的表达水平则随着蛋白质水平的升高而升高。由此表明,日粮高蛋白质水平显著降低肝脏脂肪酸合成代谢基因(ACC、FAS和SREBP-1C)的表达(P<0.05),提高脂肪酸分解代谢基因(CPT-Ⅰ和PPARα)的表达水平(P<0.05);脂肪酸合成减少,且被氧化的量增加,两者共同作用导致用于合成甘油三酯(TG)的脂肪酸含量降低,从而减少了从肝脏组织被运输到各个组织的TG含量,使机体沉积TG的能力降低。     

Study on differentiation during embryonic development across selective and ancestral breeds

In order to explore the effect of strain on diverging post‐hatch muscle properties, muscle regulation during embryo development was investigated in selected and unselected breeds. Four broiler strains were used: JingNing (JN) chicken (a Chinese native chicken), HuangYu (HY) broiler, BaiYu (BY) broiler and Hyline layer (commercial crossbred chickens). Results showed that the four breeds had almost the same characteristic during different incubation periods. BY broilers moved more than JN and Hyline layers from Hamburger & Hamilton stage (HH)24 to HH31 (P < 0.05). HY broilers moved more than JN and Hyline layers from HH27 to HH31 (P < 0.01). All the embryos were heavier daily from HH24 to ED18 (P < 0.05); broilers presented greater body weights than JN and hyline layers (P > 0.05); broilers presented smaller fiber diameter than JN chickens before HH31 (P > 0.05). From then on, JN chicken exhibited smaller fiber diameter compared to the broilers (P > 0.05). Western blotting indicated all the breeds had continuous insulin‐like growth factor‐I (IGF‐I) expression, with the highest expression level in broilers from HH19 to HH24 and highest expression level in JN chicks from HH27 to HH31. The results indicated that the diverging growth among breeds was already shown in embryonic stages; the different expression patterns of IGF‐I may be involved in cell proliferation and differentiation.     

Whole-genome bisulfite sequencing of abdominal adipose reveals DNA methylation pattern variations in broiler lines divergently selected for fatness

The methylation status of pivotal genes involved in fat deposition in chickens has been extensively studied. However, the whole-genome DNA methylation profiles of broiler abdominal adipose tissue remain poorly understood. Using whole-genome bisulfite sequencing, we generated DNA methylation profiles of chicken abdominal adipose tissue from Northeast Agricultural University broiler lines divergently selected for abdominal fat content. We aimed to explore whether DNA methylation was associated with abdominal fat deposition in broilers. The whole-genome DNA methylation profiles of fat- and lean-line broilers abdominal adipose tissue were constructed. The DNA methylation levels of functional genomic regions in the fat broiler were higher than those in the lean broiler, especially in the 3′ untranslated regions (UTRs) and exons in the non-CG contexts. Additionally, we identified 29,631 differentially methylated regions and, subsequently, annotated 6,484 and 2,016 differentially methylated genes (DMGs) in the gene body and promoter regions between the two lines, respectively. Functional annotation showed that the DMGs in promoter regions were significantly enriched mainly in the triglyceride catabolic process, lipid metabolism-related pathways, and extracellular matrix signal pathways. When the DMG in promoter regions and differentially expressed genes were integrated, we identified 30 genes with DNA methylation levels that negatively correlated with their messenger RNA (mRNA) expression, of which CMSS1 reached significant levels (false discovery rate < 0.05). These 30 genes were mainly involved in fatty acid metabolism, peroxisome-proliferator-activated receptor signaling, Wnt signaling pathways, transmembrane transport, RNA degradation, and glycosaminoglycan degradation. Comparing the DNA methylation profiles between fat- and lean-line broilers demonstrated that DNA methylation is involved in regulating broiler abdominal fat deposition. Our study offers a basis for further exploring the underlying mechanisms of abdominal adipose deposition in broilers.     

早期热应激对肉仔鸡后期生长发育的影响

旨在研究早期热应激对肉仔鸡后期生长发育及骨骼肌中相关信号通路基因表达的影响。本研究选取健康、体重相近的刚出壳AA肉仔鸡192只（公母各半）,随机分成2组（每组4个重复,每个重复24只鸡）,饲喂至3日龄时,分别给予36℃热应激处理24 h （热应激组）和正常温度（33℃）饲喂（对照组）。42日龄时每个处理选取8只鸡采取血浆,屠宰后测定胴体性能,分离胸肌样品测定骨骼肌发育有关的基因表达。结果表明,热应激当天肉仔鸡增重显著降低,但整个饲养期肉仔鸡的平均日增重显著增加（P<0.05）;与对照组相比,早期热应激有提高整个饲养期肉仔鸡平均日采食量的趋势（0<P<0.1）。早期热应激对肉仔鸡的屠宰率、半净膛重、全净膛重、肝比重、心比重、胸肌率、腿肌率、腹脂率都无显著影响（P>0.05）。早期热应激显著升高了42日龄肉仔鸡血浆酮体含量（P<0.05）。在胸肌中,早期热应激显著提高了胰岛素样生长因子（IGF-1）及其受体（IGF-1R）的基因表达（P<0.05）。本试验结果提示,早期热应激促进了肉仔鸡后期的生长发育,骨骼肌中IGF-1信号可能参与此调节过程。     

Nutrition and metabolism in poultry: role of lipids in early diet

Modern strains of broiler chickens are selected for fast growth and are marketed anywhere from 36 to 49 days after a21-day incubational period. For a viable healthy chick, all the necessary nutrients required for growth and development must be provided by the hen through the fertilized egg. The current feeding strategies for improved growth, health and productivity are targeted towards chicks after hatching. Considering the fact that developing chick embryo spends over 30 % of its total life span inside the hatching egg relying on nutrients deposited by the breeder hen, investigations on nutritional needs during pre-hatch period will improve embryonic health, hatchability and chick viability. In this context, investigations on hatching egg lipid quality is of utmost importance because, during incubation, egg fat is the major source of energy and sole source of essential omega-6(n-6) and omega-3(n-3) fatty acids to the chick embryo.Due to the unique roles of n-3 and n-6 fatty acids in growth, immune health, and development of central nervous system, this review will focus on the role of early exposure to essential fatty acids through maternal diet and hatching egg and its impact on progeny in meat-type broiler chickens.     

Corticosterone administration and high-energy feed results in enhanced fat accumulation and insulin resistance in broiler chickens

1. Two experiments were conducted to investigate the effects of exogenous corticosterone administration (30 mg/kg diet) and dietary energy level on feed or energy intake and fat deposition in broiler chickens of 1 and 4 weeks of age. 2. Corticosterone treatment significantly suppressed body weight (BW) gain and reduced feed and caloric efficiencies. The retarded growth may conceal the stimulatory effect of corticosterone on feed consumption or metabolisable energy (ME) intake. A high-energy diet may increase energy intake and partially alleviate the suppressing effect of corticosterone on growth of broilers. 3. Corticosterone administration promoted the conservation of energy stores as fat at both abdominal and subcutaneous sites and this process occurred regardless of dietary energy level in ad libitum feeding status. A high-energy diet increased fat accumulation and showed no significant interaction with corticosterone treatment. 4. The suppressed development of breast and thigh muscles by corticosterone treatment was observed only in 1-week-old chickens fed on the low-energy diet. In contrast, the yield of breast muscle but not thigh muscle was significantly decreased by corticosterone in 4-week-old chickens, suggesting that the tissue specificity to corticosterone challenge is age dependent. 5. Plasma concentrations of glucose, insulin, triglyceride, non-esterified fatty acids (NEFA) and very low density lipoprotein were increased by corticosterone treatment regardless of diet treatment. A high-energy diet increased plasma levels of NEFA and resulted in hyperinsulinism in 4-week-old chickens but not in 1-week-old chickens. 6. Lipoprotein lipase (LPL) activities in adipose tissues may have been up-regulated by corticosterone treatment and showed tissue specificity. The increased LPL activities at ad libitum feeding status were not necessarily linked with the increased fat accumulation in corticosterone challenged chickens. 7. Corticosterone resulted in augmented energy consumption and altered energy redistribution toward lipid deposition. The induced insulin resistance and enhanced hepatic de novo lipogenesis by corticosterone are likely to be responsible for the increased fat deposition.     

混饮低剂量ChIFN-α对健康肉鸡免疫功能及生产性能的影响

试验选取50只雏肉鸡饲养至14日龄,随机分为5组,每组10只, 14～42日龄期间试验Ⅱ、Ⅲ、Ⅳ组每日分别混饮0.1、1.0、10 U/mL剂量ChIFN-α（鸡干扰素-α）,Ⅰ组设为空白组,混饮自来水。Ⅴ组设为阳性对照,混饮50 mg/L盐酸左旋咪唑,研究混饮ChIFN-α后对肉鸡免疫功能及生产性能的影响。结果显示,混饮ChIFN-α后可提高肉鸡外周血白细胞总数、淋巴细胞百分比和嗜碱性粒细胞百分比,降低嗜中性粒细胞百分比和单核细胞百分比;提高肉鸡脾脏指数及血清总蛋白(TP)、血清白蛋白(ALB)、血清球蛋白(GLO)的浓度及ALB/GLO比值;不影响肉鸡的生长发育。因此,混饮低剂量的ChIFN-α可提高肉鸡的免疫功能,对健康肉鸡的生长无显著的影响。     

DHA对高脂饲粮诱导肝脏脂肪积累的预防作用

[目的] 探索二十二碳六烯酸（DHA）对高脂饲粮诱导肝脏脂肪积累的预防机制。[方法] 将32只雄性SPF级C57BL/6小鼠均分为4组,对照组（Con）饲喂普通饲粮,模型组（Model）饲喂高脂饲粮,DHA组分别在高脂饲粮中添加0.2 g DHA（DHAL）和1.0 g DHA（DHAH）,饲喂周期为20周。饲喂期间每天称量体重及食物重量,计算摄食量;饲喂结束,采集肝脏和血液,ELISA法检测肝脏脂联素和血清甘油三酯（TG）的含量,实时荧光定量PCR检测新生脂肪合成关键酶（SREBP-1c、FAS）、脂肪酸氧化关键基因（PPARα、PPARγ、CPT-1A和ACOX）、线粒体基因（PGC-1α）、褐色脂肪化基因（Prdm16、UCP1）的表达,Western blotting法检测肝脏磷酸化ACC、AMPK和AKT蛋白的表达。[结果] 与Con组相比,Model组终体重、体脂重量和TG含量均显著增加（P<0.05）,肝脏脂联素浓度显著降低（P<0.05）。与Model组相比,DHAL和DHAH组终体重、体脂重量和TG含量均显著降低（P<0.05）,肝脏脂联素水平显著增加（P<0.05）。实时荧光定量PCR结果表明,与Con组相比,Model组SREBP-1c和FAS mRNA表达量均显著增加（P<0.05）,PPARα、CPT-1A、ACOX、PGC-1α和UCP1 mRNA表达均显著降低（P<0.05）;与Model组相比,DHAL和DHAH组SREBP-1c和FAS mRNA表达量均显著降低（P<0.05）,且DHAH组表达量均显著低于DHAL组（P<0.05）,DHAL和DHAH组PPARα、CPT-1A、ACOX、PGC-1α、Prdm16和UCP1 mRNA表达量均显著增加（P<0.05）,DHAH组CPT-1A和ACOX mRNA表达量显著低于DHAL组（P<0.05）,PPARγ的mRNA表达量在4组中没有显著差异（P>0.05）。Western blotting结果表明,Model组p-ACC显著高于Con和DHAH组,但显著低于DHAL组;Model组p-AMPK/AMPK和pAKT/AKT比值均显著低于Con组（P<0.05）,DHAH组p-AMPK/AMPK和pAKT/AKT比值均显著高于Model和DHAL组（P<0.05）。[结论] DHA可降低高脂饲粮导致的C57BL/6小鼠终体重、体脂和TG含量的升高,增加肝脏脂联素的水平,促进脂肪酸氧化和白色脂肪细胞褐色化,从而预防肝脏的脂肪积累。     

Nutrition and metabolism in poultry: role of lipids in early diet

Modern strains of broiler chickens are selected for fast growth and are marketed anywhere from 36 to 49 days after a 21-day incubational period. For a viable healthy chick, all the necessary nutrients required for growth and development must be provided by the hen through the fertilized egg. The current feeding strategies for improved growth, health and productivity are targeted towards chicks after hatching. Considering the fact that developing chick embryo spends over 30 % of its total life span inside the hatching egg relying on nutrients deposited by the breeder hen, investigations on nutritional needs during pre-hatch period will improve embryonic health, hatchability and chick viability. In this context, investigations on hatching egg lipid quality is of utmost importance because, during incubation, egg fat is the major source of energy and sole source of essential omega-6 (n-6) and omega-3 (n-3) fatty acids to the chick embryo. Due to the unique roles of n-3 and n-6 fatty acids in growth, immune health, and development of central nervous system, this review will focus on the role of early exposure to essential fatty acids through maternal diet and hatching egg and its impact on progeny in meat-type broiler chickens.     

Up-regulation of Endothelin-1 and Endothelin type A receptor genes expression in the heart of broiler chickens versus layer chickens

To compare Endothelin (ET) production and genes expression of ET-1 and ET_A receptor (ET_AR) between broiler and layer chickens during rearing, semi-quantitative RT-PCR and enzyme immunometric assay were performed in the heart ventricles and serum. There were gradual elevations of ET-1 and ET_AR mRNAs in the left ventricle of broiler and layer chicken groups that were mainly significant (P < 0.05) at 28, 35 and 42 days of age with compared to previous days whereas were not significant between two groups.These gradual elevations of ET-1 and ET_AR mRNAs were also observed in the right ventricle that were significant (P < 0.05) at 28, 35 and 42 days of age in broilers and 42 days of age in layers with compared to previous days. Increasing of these mRNAs in the right ventricle of broiler chickens were significantly (P < 0.05) more than layer chickens at 28, 35 and 42 days. Serum ET in broilers was significantly (P < 0.05) higher than layer chickens at 28 and 42 days of age. It is concluded that circulating ET and cardiac ET-1, ET_AR genes expression is higher in broiler chickens than in layer chickens particularly after 21 days of age. It is probably that these breed differences make broiler chickens to be more susceptible to Endothelin related-cardiomyopathies such as congestive heart failure and ascites.