Myostatin down‐regulates the IGF‐2 expression via ALK‐Smad signaling during myogenesis in cattle

Myostatin (MSTN) is a negative regulator during muscle differentiation, whereas insulin‐like growth factors (IGFs) are essential for muscle development. MSTN and IGFs act oppositely during myogenesis, but there is little information on the mutual relationship of MSTN and IGFs. The present study was conducted to examine whether MSTN affects IGF expression during early myogenesis in cattle. IGF‐1 mRNA was similarly expressed in M. longissimus thoracis of double‐muscled (DM) and normal (NM) Japanese shorthorn cattle. IGF‐2 mRNA expression was consistently higher in the normal and regenerating muscle of DM cattle than those of NM cattle. When myoblasts were isolated from regenerating M. longissimus thoracis, IGF‐2 mRNA expression showed a significant increase in differentiating DM derived myoblasts (DM‐myoblasts) as compared with differentiating NM derived myoblasts (NM‐myoblasts). An addition of recombinant mouse myostatin (rMSTN) to myoblast cultures attenuated IGF‐2 mRNA expression and decreased myotube formation, but did not effect IGF‐1 mRNA expression. An activin‐like kinase (ALK) inhibitor, SB431542, mediates MSTN action, suppressed the translocation of Smad2/3 into the nucleus in DM‐myoblasts, and restored the attenuated IGF‐2 mRNA expression and the decreased myotube formation induced by rMSTN in myoblast cultures. The findings indicate that MSTN may negatively regulate myoblast differentiation by suppressing IGF‐2 expression via ALK‐Smad signaling.     

Effects of leptin and adiponectin on the growth of porcine myoblasts are associated with changes in p44/42 MAPK signaling

We hypothesized that both adiponectin and leptin affect the growth of porcine skeletal muscle cells, with fatty acids acting as modifiers in adipokine action and that both adipokines influence the gene expression of their receptors. Therefore, the objective of this study was to investigate the effects of recombinant adiponectin and leptin on cell number (DNA) and DNA synthesis rate with and without oleic acid supplementation, on cell death, and on key intracellular signaling molecules of proliferating porcine myoblasts in vitro. Moreover, the mRNA expression of genes encoding for the leptin and adiponectin receptors (LEPR, ADIPOR1, ADIPOR2) as affected by leptin or adiponectin was examined. Recombinant porcine adiponectin (40 μg/mL) and leptin (20 ng/mL) increased DNA synthesis rate, measured as [³H]-thymidine incorporation (P < 0.01), reduced cell viability in terms of lactate dehydrogenase release (P < 0.05), or lowered DNA content after 24 h (P < 0.05). In adiponectin-treated cultures, oleic acid supplementation increased DNA synthesis rate and reduced cell number in a dose-dependent manner (P < 0.05). Both adiponectin (P = 0.07) and leptin (P < 0.05) induced a transient activation of p44/42 mitogen-activated protein kinase (MAPK) after 15 min, followed by decreases after 60 and 180 min (P < 0.05). Adiponectin tended to increase c-fos activation (P = 0.08) and decreased p53 activation at 180 min (P = 0.03). Both adiponectin and leptin down-regulated the abundance of ADIPOR2 mRNA and, transiently, of LEPR mRNA (P < 0.05). In conclusion, adiponectin and leptin may adversely affect the growth of porcine myoblasts, which is related to p44/42 MAPK signaling and associated with changes in ligand receptor gene expression.     

Differential gene expression of Eph‐ephrin A1 and LEPR‐LEP with high or low number of embryos in pigs during implantation

The objective of this study was to ascertain whether mRNA and protein expressions of implantation‐related genes (erythropoietin‐producing hepatocellular receptor–ligand A1, Eph‐ephrin A1 and leptin receptor–leptin, LEPR‐LEP) differed between pigs with high and low number of embryos, and whether these differences in gene expression might affect embryo implantation. Experimental pig groups (n = 24) for high and low number of embryos were prepared by altering the number of eggs ovulated in pre‐pubertal gilts treated with 1.5 × (High) or 1.0 × (Low) PG600 ([400 IU PMSG + 200 IU hCG]/dose, AKZO‐NOBEL). Gilts expressing oestrus were artificially inseminated twice and maintained in breeding and gestation until the reproductive tract was collected on day 22 of pregnancy. At slaughter, the reproductive tracts from each pregnant gilt from each treatment were immediately processed to collect samples for RNA and protein analysis. Within each gilt, three conceptus points were sampled, one from each horn and then a random conceptus within the tract. At each conceptus point, endometrial attachment site, chorion–allantois and embryo were collected and immediately frozen in liquid nitrogen. Number of corpus luteum (CL) (35.4 vs. 12.6) and total embryo number (18.8 vs. 10.2) were greater in the high‐embryo compared to the low‐embryo group, respectively (p < .05). Real‐time qPCR results showed that Eph‐ephrin A1 mRNA expression was less in the high‐embryo (p < .05) compared to the low‐embryo group. In addition, Western blotting analysis indicated that Eph‐ephrin A1 and LEP protein expression at endometrial attachment site in high‐embryo was less (p < .05) compared to low‐embryo group. It was also noted that mRNA expression of Eph‐ephrin A1 and LEPR‐LEP was greater in pregnant than non‐pregnant gilts (p < .05). Moreover, mRNA expression of Eph‐ephrin A1 (p < .05) and LEPR‐LEP was greatest at endometrial attachment site among all three tissues. There was a positive correlation between expressions of Eph‐ephrin A1, LEPR‐LEP and embryo length with the correlation coefficient 0.31–0.59. For Eph‐ephrin A1, the highest correlation coefficient appeared between Eph A1 expression and normal embryo number, between ephrin A1 expression and embryo length. For LEPR‐LEP, the highest correlation coefficient appeared between LEPR‐LEP expression and ovary weight (0.79 for both, p < .05), followed by embryo length and weight. The results of this study suggest that low expression of Eph‐ephrin A1 and LEPR‐LEP is somehow related to increased embryo number during implantation and that endometrial attachment site might be the main target tissue of these gene products. Yet, the increased expression of Eph‐ephrin A1 and LEPR‐LEP appeared associated with increased embryo growth (length and weight) and ovary weight, Eph‐ephrin A1 and LEPR‐LEP might play roles in the regulation of embryo implantation in pigs.     

Lysine restriction and realimentation affected growth,blood profiles and expression of genes related to protein and fat metabolism in weaned pigs

To investigate the effects of lysine restriction and subsequent realimentation on growth performance, blood profiles and gene expression of leptin and myostatin, 128 weaned pigs [initial body weight (BW) 6.96 ± 1.07 kg, 26 ± 2 days of age] were randomly allotted to four treatments. The starter diets during the first 2 weeks (P1) contained 100%, 80%, 70% or 60% of recommended lysine levels ( National Research Council, 1998 ). Then, common grower 1 and 2 diets were offered for 2 weeks (P2 and P3) each. During P1, average daily gain (ADG) was linearly reduced (p < 0.05) with the increasing levels of lysine restriction. Growth rate was greater in pigs previously fed lysine‐restricted diets than well‐fed pigs although it did not reach a significant level during realimentation. However, the final BW and overall ADG were the lowest (p < 0.05) and F/G was poor in pigs fed 60% lysine diet. Relative visceral organ weights and composition of skeletal muscle were similar (p > 0.05) among the treatment. Blood triglyceride and glucose levels were increased (p < 0.05) during P1, while blood urine nitrogen, total protein and albumin levels were decreased (p < 0.05) during P2 with the reduction in dietary lysine levels. The abundance of myostatin mRNA in skeletal muscle and leptin mRNA in subcutaneous adipose tissue were lower (p < 0.05) in lysine‐restricted pigs than in pigs fed non‐restricted diets. In conclusion, 80% and 70% lysine restriction of starter diets resulted in inferior growth and compensatory growth effect was noted during realimentation, while 60% lysine restriction had a negative influence on growth performance. Moreover, the changes in myostatin and leptin mRNA abundance caused by nutritional manipulations may be involved in the regulation of protein and fat deposition in young pigs.     

Effect of dietary methionine on growth performance and insulin‐like growth factor‐I mRNA expression of growing meat rabbits

An experiment was conducted to determine the effects of different amounts of dietary methionine on growth performance, serum protein, growth hormone (GH), insulin‐like growth factor‐I (IGF‐I) concentrations and IGF‐I mRNA expression of growing meat rabbits. One hundred weaned growing meat rabbits were allocated to individual cages and randomly divided into five groups. The methionine addition concentrations of the five groups were 0, 2, 4, 6 and 8 g/kg diet (as‐fed basis) and sulphur amino acids (SAA) concentrations ranging from 3.8 to 11.6 g/kg diet, respectively. The results obtained were as follows: the average daily gain of 2, 4 and 6 g/kg diet groups was higher than that of 0 g/kg diet group (p < 0.01). The feed gain ratio of the 4 g/kg diet group was lower than those of 0 and 8 g/kg diet group (p < 0.01). Methionine concentrations did not affect serum urea nitrogen, total protein, insulin and IGF‐I concentration (p > 0.05). The quadratic effects of methionine on the serum concentration of albumin (Alb) and GH were obtained (p = 0.013, p = 0.018). The quadratic effect of methionine amount on IGF‐I mRNA expression was obtained (p = 0.045). The serum concentration of Alb of the 4 g/kg diet group was higher than those of 0 and 8 g/kg diet group (p < 0.01). The serum concentration of GH of 8 g/kg diet group was higher than that of the 0 g/kg diet group (p < 0.05). The liver IGF‐I mRNA expression of 4 g/kg diet group was higher than those of the 0 and 8 g/kg diet group (p < 0.05). Providing a diet mainly consisted of corn, wheat bran and peanut vine, the optimum dietary methionine addition concentration and SAA concentration for a weaner to 2‐month‐old growing meat rabbits were shown to be 2 and 5.7 g/kg diet respectively.     

Polymorphism in promoter region of growth hormone receptor is associated with potential production capacity of insulin‐like growth factor‐1 in pre‐pubertal Holstein heifers

Insulin‐like growth factor‐1 (IGF‐1) is one of the important factors for growth, milk production and reproductive functions and mainly released from the liver in response to growth hormone (GH) via GH receptor (GHR) in cattle. Recently, some single nucleotide polymorphisms (SNPs) were identified in the bovine GHR gene. Some GHR‐SNPs were shown to be related to plasma IGF‐1 concentration in cattle. Hence, the capacity to IGF‐1 production in the liver might be affected by GHR‐SNP and associated with performance in the future. This study examined whether GHR‐SNP is associated with IGF‐1 production in the liver of pre‐pubertal heifers. In 71 Holstein calves, blood samples for genomic DNA extraction were obtained immediately after birth. To genotype the GHR‐SNPs in the promoter region, polymerase chain reaction (PCR) products were digested with restriction enzyme NsiI (cutting sites: AA, AG and GG). All heifers at 4 months of age were intramuscularly injected with 0.4 mg oestradiol benzoate. Blood samples were obtained from the jugular vein just before (0 h) and 24 h after injection. The number of AA, AG and GG at the NsiI site was 0, 17 and 54 respectively. In AG and GG, plasma GH concentrations were higher pre‐injection than 24 h post‐injection (p < 0.01). Moreover, plasma GH concentrations in AG post‐injection were higher than in GG (p < 0.05). In contrast, the GG genotype exhibited higher plasma IGF‐1 concentrations in pre‐injection than post‐injection (p < 0.01), although oestradiol did not change IGF‐1 concentration in the AG genotype. We conclude that the GG polymorphism in the promoter region of GHR is associated with a higher potential capacity of IGF‐1 production in the liver of cattle.     

Effects of dietary supplementation with betaine on muscle growth,muscle amino acid contents and meat quality in Cherry Valley ducks

The effects of dietary betaine supplementation on growth performance, carcass characteristics, muscle amino acid contents, meat quality, antioxidant capacity, myogenic gene expression and mechanistic target of rapamycin (mTOR) signalling pathway in Cherry Valley ducks were evaluated. A total of 720 1‐day‐old Cherry Valley ducks were randomly distributed into four groups with six replicates of 30 birds for a 42‐day feeding trial. Ducks were fed a basal diet supplemented with 0 (control), 250, 500 or 1,000 mg/kg betaine, respectively. Growth performance was not affected by betaine. Incremental levels of betaine linearly (p < 0.05) increased the breast muscle yield and linearly (p < 0.05) decreased the subcutaneous fat thickness and the abdominal fat yield. The contents of methionine, serine, glycine, glutamate and total non‐essential amino acid in breast muscle were linearly (p < 0.05) increased by betaine supplementation. With increasing betaine levels, the drip loss and the content of malondialdehyde (MDA) were linearly (p < 0.05) decreased, and the redness of meat (linear p < 0.05), the activities of catalase (CAT) (linear p < 0.05) and total superoxide dismutase (T‐SOD) (linear p < 0.05, quadratic p < 0.05) were increased. Moreover, the myogenic differentiation factor 1 (MyoD1) mRNA expression and the mTOR mRNA expression and protein phosporylation were linearly (p < 0.05) up‐regulated, and the myostatin (MSTN) mRNA expression was linearly (p < 0.05) down‐regulated by betaine supplementation. Overall, this study indicated that betaine supplementation did not affect the growth performance of Cherry Valley ducks, but could linearly increase some amino acid contents in breast muscle, especially glycine, and increase muscle antioxidant activity to improve meat quality. Moreover, betaine supplementation could improve the breast muscle yield by increasing MyoD1 mRNA expression, decreasing MSTN mRNA expression and regulating mTOR signalling pathway.     

Effect of genistein on IGF‐1 and IGFBP‐1 in young and aged female rat ovary

The objective of this study was to assess the effects of genistein (GEN) on expression of insulin‐like growth factor 1 (IGF‐1) and insulin‐like growth factor binding protein 1 (IGFBP‐1) in young and aged rat ovary. Forty young female Sprague Dawley (SD) rats (200 ± 20 g) and forty aged female SD rats (490 ± 20 g) were selected and according to weight, they were divided into the following five groups with eight animals in each: negative control group (NC), low‐dose group (L), middle‐dose group (M), high‐dose group (H) and positive control group (PC). GEN group received GEN of 15, 30, 60 mg/kg respectively. It lasted 30 days. Concentrations of serum hormones, IGF‐1 and IGFBP‐1 were determined by enzyme‐linked immunosorbent assay (ELISA). Gene and protein expressions of IGF‐1 and IGFBP‐1 were determined by real‐time PCR and Western blot respectively. Compared with NC, GEN significantly increased oestradiol‐17β(E₂) level in aged rat, reduced luteinizing hormone (LH) level in young and aged rat. Serum levels of IGFBP‐1 in young rats were significantly higher in GEN groups (p < 0.05). mRNA and protein expression levels of IGF‐1 and IGFBP‐1 were positively correlated with GEN dose. GEN could significantly reduce the ratio of IGF‐1/IGFBP‐1 of aged rats. Multivariate Cox regression analysis result showed IGF‐1 and IGFBP‐1 levels significantly correlated with GEN dose. We speculate that there is an association between the addition of GEN and expression of IGF‐1 and IGFBP‐1, and the relationship between them is different in young and aged rat.     

Effects of breed and dietary nutrient density on the growth performance,blood metabolite,and genes expression of target of rapamycin (TOR) signalling pathway of female broiler chickens

This study was conducted to compare the effects of exchanged diets with identical energy level on characteristics of slow‐growing (WENs Yellow‐Feathered Chicken, WYFC) and fast‐growing (White Recessive Rock Chicken, WRRC) female chickens. A total of 1450 WYFC and 1150 WRRC 1‐day‐old female hatchlings were used. A high‐nutrient‐density (HND) diet and a low‐nutrient‐density (LND) diet were formulated for three phases. A completely randomized experimental design with a 2 × 2 factorial arrangement (diet and breed), each with five replicates of 145 and 115 birds, was applied. The results showed that WRRC had a higher body weight (BW), average daily feed intake and average daily gain than WYFC throughout the experiment (p < 0.05). WYFC that were provided with HND groups had a higher BW only in the starter and grower phases, whereas WRRC had a higher BW in the HND group than in LND groups throughout the experiment. The feed:gain ratio and protein efficiency ratio (PER) were better for WRRC in the starter and grower phases; however, these ratios were better for WYFC in the finisher period. The LND groups had a higher PER throughout the experiment for both breeds (p < 0.05). The breast and leg muscle weights were higher for WRRC compared with WYFC during the grower and finisher phases (p < 0.05). WRRC had a lower liver index but higher serum UA and alkaline phosphatase (ALP) concentrations than WYFC (p < 0.05). No diet effect was observed on organ indices, muscle yields or blood responses. The gene expressions of Rheb, TOR, S6K1 and 4E‐BP1 in gastrocnemius muscle were the highest in the WYFC‐LND groups at 63 and 105 days (p < 0.05). These findings suggested that different genotypes respond differently to changes in dietary nutrient density and that lower‐nutrient‐density diets are optimal for the long‐term housing of broiler chickens.     

Effects of colostrum feeding and dexamethasone treatment on mRNA levels of insulin-like growth factors (IGF)-I and -II, IGF binding proteins-2 and -3, and on receptors for growth hormone, IGF-I, IGF-II, and insulin in the gastrointestinal tract of neonatal calves

The somatotropic axis regulates growth of the gastrointestinal tract (GIT). In addition, colostrum feeding and glucocorticoids affect maturation of the GIT around birth in mammals. We have measured mRNA levels of members of the somatotropic axis to test the hypothesis that colostrum intake and dexamethasone treatment affect respective gene expression in the GIT. Calves were fed either colostrum or an isoenergetic milk-based formula, and in each feeding group, half of the calves were treated with dexamethasone (DEXA; 30 microg/kg body weight per day). Individual parameters of the somatotropic axis differed (P < 0.05) among different GIT sections and formula feeding increased (P < 0.05) mRNA levels of individual parameters at various sites of the GIT. Effects of DEXA on the somatotropic axis in the GIT partly depended on different feeding. In colostrum-fed calves, DEXA decreased (P < 0.05) mRNA levels of IGF-I (esophagus, fundus, duodenum, and ileum), IGF-II (fundus), IGFBP-2 (fundus), IGFBP-3 (fundus), IGF1R (esophagus, ileum, and colon), IGF2R (fundus), GHR (fundus), and InsR (esophagus, fundus), but in formula-fed calves DEXA increased mRNA levels of IGF-I (esophagus, rumen, jejunum, and colon). Furthermore, DEXA increased (P < 0.05) mRNA levels of IGF-II (pylorus), IGFBP-3 (duodenum), IGF2R (pylorus), and GHR (ileum), but decreased mRNA levels of IGFBP-2 (ileum), and IGF1R (fundus). Whereas formula feeding had stimulating effects, effects of DEXA treatment on the gene expression of parameters of the somatotropic axis varied among GIT sites and partly depended on feeding.     

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.     

籽鹅和莱茵鹅MSTN和MyoG基因表达及其与屠宰性状的相关分析

本研究旨在探讨肌肉生成抑制素(myostatin,MSTN)和肌细胞生成素(myogenin,MyoG)基因对鹅骨骼肌生长发育的影响。试验以莱茵鹅和籽鹅为研究对象,采用实时荧光定量PCR方法测定MSTN、MyoG基因在籽鹅和莱茵鹅胸肌、腿肌中的差异表达情况,运用统计软件对基因表达情况与屠宰性状间的相关性进行分析。结果显示,莱茵鹅胸肌重和胸肌率极显著高于籽鹅(P<0.01),籽鹅胸肌MSTN、MyoG mRNA表达量极显著高于莱茵鹅(P<0.01);在腿肌中,籽鹅MSTN mRNA表达量极显著低于莱茵鹅(P<0.01),籽鹅与莱茵鹅MyoG mRNA表达量间无显著差异(P>0.05)。同一品种MSTN mRNA表达水平也存在差异,在籽鹅胸肌和腿肌中表达量存在极显著差异(P<0.01);在莱茵鹅胸肌和腿肌中表达量存在显著差异(P<0.05)。就MyoG mRNA而言,在籽鹅和莱茵鹅胸肌和腿肌中表达量存在极显著差异(P<0.01)。MSTN基因表达与屠宰性能相关性分析表明,胸肌中MSTN mRNA表达量与活重、胸肌重和胸肌率呈极显著负相关(P<0.01);胸肌和腿肌中MyoG mRNA表达量与活重呈极显著正相关(P<0.01)。说明MSTN和MyoG基因可能对肌肉生长分别起正负调控作用。     

Expression of MSTN and MyoG Genes in Zi and Rhine Goose and their Correlation with Carcass Traits

The aim of this study was to investigate the effects of MSTN and MyoG genes on goose skeletal muscle growth.In this study,MSTN and MyoG genes expression were detected in breast and leg muscle of Zi and Rhine goose by Real-time PCR,and the correlations between genes expression levels and carcass traits were investigated.The results showed that the breast muscle weight and breast muscle rate of Rhine goose were extremely significant higher than Zi goose (P<0.01).MSTN and MyoG mRNA expression in breast muscle of Zi goose were significantly higher than that of Rhine goose,and the mRNA level of MSTN in leg muscle of Rhine was extremely significant higher than that of Zi goose (P<0.01),there was no significant difference of MyoG mRNA between Zi goose and Rhine goose (P>0.05).There was extremely significant difference between MSTN mRNA expression in breast muscle and leg muscle of Zi goose (P<0.01).MSTN mRNA expression in leg muscle was significantly higher than that of breast muscle of Rhine goose (P<0.05).There was extremely significant difference between MyoG mRNA expression in breast muscle and leg muscle of Zi goose and Rhine goose (P<0.01).There was a extremely significant negative correlation between MSTN mRNA expression in breast muscle with body weight,breast muscle weight and breast muscle percentage (P<0.01).There was a extremely significant positive correlation between MyoG mRNA expression in breast muscle and leg muscle with body weight (P<0.01).MSTN and MyoG gene might have positive and negative regulation effect on muscle growth.     

In ovo L‐arginine supplementation stimulates myoblast differentiation but negatively affects muscle development of broiler chicken after hatching

In this study, we tested the hypothesis that in ovo feeding (IOF) of L‐arginine (L‐Arg) enhances nitric oxide (NO) production, stimulates the process of myogenesis, and regulates post‐hatching muscle growth. Different doses of L‐Arg were injected into the amnion of chicken embryos at embryonic day (ED) 16. After hatching, the body weight of individual male chickens was recorded weekly for 3 weeks. During in vitro experiments, myoblasts of the pectoralis major (PM) were extracted at ED16 and were incubated in medium containing 0.01 mm L‐Arg, 0.05 mm L‐Arg, and (or) 0.05 mm L‐nitro‐arginine‐methyl‐ester (L‐NAME), an inhibitor of nitric oxide synthase (NOS). When 25 mg/kg L‐Arg/initial egg weight was injected, no difference was observed in body weight at hatch, but a significant decrease was found during the following 3 weeks compared to that of the non‐injected and saline‐injected control, and this also affected the growth of muscle mass. L‐NAME inhibited gene expression of myogenic differentiation antigen (MyoD), myogenin, NOS, and follistatin, decreased the cell viability, and increased myostatin (MSTN) gene expression. 0.05 mm L‐Arg stimulated myogenin gene expression but also depressed muscle cell viability. L‐NAME blocked the effect of 0.05 mm L‐Arg on myogenin mRNA levels when co‐incubated with 0.05 mm L‐Arg. L‐Arg treatments had no significant influence on NOS mRNA gene expression, but had inhibiting effect on follistatin gene expression, while L‐NAME treatments had effects on both. These results suggested that L‐Arg stimulated myoblast differentiation, but the limited number of myoblasts would form less myotubes and then less myofibers, while the latter limited the growth of muscle mass.     

Correlation analysis of hypothalamic mRNA levels of appetite regulatory neuropeptides and several metabolic parameters in 28‐day‐old layer chickens

Various lines of evidence suggest that appetite‐related neuropeptides in the hypothalamus are regulated by adiposity signals such as leptin and insulin in mammals. In the present study, we examined age‐dependent changes in the weight of abdominal fat and hypothalamic mRNA levels of neuropeptide Y (NPY, an orexigenic neuropeptide) and proopiomelanocortin (POMC, a precursor of anorexigenic neuropeptides) in growing chickens at 7, 14, 21 and 28 days of age. Hypothalamic NPY mRNA levels were significantly (P < 0.05) decreased after 14 days of age, whereas hypothalamic POMC mRNA levels were significantly (P < 0.05) increased at 28 days of age. The percentage of abdominal fat was significantly increased after 14 days of age in chickens. We next examined the correlation of hypothalamic NPY and POMC mRNA levels and several parameters at 28 days of age. There were no significant correlations between hypothalamic mRNA levels of NPY or POMC and the percentage of abdominal fat. These findings suggest that the gene expressions of NPY and POMC do not depend on adiposity in chickens, at least in 28‐day‐old layer chickens.     

Influence of recombinant duck follistatin protein on embryonic muscle development and gene expressions

Follistatin (FST) acts as a positive regulator of muscle development by inhibiting the activities and expression of myostatin. The recombinant duck FST protein was injected into hatching eggs and was also added to the medium of duck myoblast to study its role on duck embryonic muscle development and gene expressions. Duck embryo weight increased 3.49% (p > 0.05) in FST treatment group as compared with control group, but minor effects were found on leg or breast muscle weights of ducklings at 2 days post‐hatching (p > 0.05). Relative expression of Pax7 was upregulated in both leg and breast muscle tissues (p < 0.05), while MyoD was only upregulated in leg muscle (p < 0.05), and Myf5 was only upregulated in breast muscle (p < 0.05). Relative expression of myostatin was downregulated in both muscle tissues researched (p < 0.05). In vitro studies also showed some maker genes relevant to protein synthesis and degradation, cells’ proliferation and differentiation had significant changes in myoblasts after treated with FST. These results suggested that in ovo feeding of recombinant FST protein to duck hatching eggs had an effect on duck embryo development but have less roles on the duck embryonic muscle development.