Differentiation capacities of skeletal muscle satellite cells in Lantang and Landrace piglets.
ABSTRACT: We isolated and cultured satellite cells (SCs) from the longissimus dorsi muscles of 1-day-old male Landrace and Lantang piglets to compare the SC differentiation capacity in the two breeds. Lantang piglets yielded more (P < 0.05) SCs per gram of muscle than Landrace piglets (5.2 ± 0.9×104 vs. 2.4 ± 0.2×104). Transcription of the differentiation markers myogenin and myosin heavy chain I (MyHC I) in the longissimus dorsi muscle was higher in Lantang than Landrace piglets (P < 0.05). Protein levels of myogenin (P < 0.05), MyHC I (P < 0.05), and myogenic regulatory factor 4 (P = 0.07) were higher in Lantang than Landrace piglet SCs after 72 h of differentiation. Creatine kinase activity was higher in Lantang than Landrace piglet SCs after 24, 48, and 72 h of differentiation (P < 0.05), and there was a greater fusion index in Landrace piglet SCs after 72 h of differentiation. In addition, phosphorylation of Akt, mTOR, S6K1, S6, and 4EBP1 was lower in Lantang than Landrace piglet SCs (P < 0.05). Thus differentiation was more extensive in Lantang than Landrace piglet SCs, but expression of the mTOR signaling pathway was lower in Lantang piglet SCs, suggesting mTOR signaling may inhibit myogenic differentiation. These findings reveal that mTOR signaling is a factor in myogenesis and imply that mTOR could potentially serve as an activator of myoblast differentiation and muscle regeneration.
Project description:Inulin is one of the commercially feasible dietary fibers that has been implicated in regulating the gut health and metabolism of animals. This experiment was conducted to investigate the effect of dietary inulin supplementation on growth performance and meat quality in growing-finishing pigs. Thirty-six Duroc × Landrace × Yorkshire White growing barrows (22.0 ± 1.0 kg) were randomly allocated to two dietary treatments consisting of a basal control diet (CON) or basal diet supplemented with 0.5% inulin (INU). Results showed that inulin supplementation tended to increase the average daily gain (ADG) at the fattening stage (0.05 < p < 0.10). Inulin significantly increased the dressing percentage (p < 0.05) and tended to increase the loin-eye area. The serum concentrations of insulin and IGF-I were significantly higher (p < 0.05) in the INU group than in the CON group. Moreover, inulin supplementation significantly elevated the expression level of myosin heavy chain II b (MyHC IIb) in the longissimus dorsi (p < 0.05). Inulin significantly upregulated the expression of mammalian rapamycin target protein (mTOR) but decreased (p < 0.05) the expression level of muscle-specific ubiquitin ligase MuRF-1. These results show the beneficial effect of inulin supplementation on the growth performance and carcass traits in growing-finishing pigs, and will also facilitate the application of inulin in swine production.
Project description:There is little genomic information regarding gene expression differences at the whole blood transcriptome level of different pig breeds at the neonatal stage. To solve this, we characterized differentially expressed genes (DEGs) in the whole blood of Dapulian (DPL) and Landrace piglets using RNA-seq (RNA-sequencing) technology. In this study, 83 DEGs were identified between the two breeds. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses identified immune response and metabolism as the most commonly enriched terms and pathways in the DEGs. Genes related to immunity and lipid metabolism were more highly expressed in the DPL piglets, while genes related to body growth were more highly expressed in the Landrace piglets. Additionally, the DPL piglets had twofold more single nucleotide polymorphisms (SNPs) and alternative splicing (AS) than the Landrace piglets. These results expand our knowledge of the genes transcribed in the piglet whole blood of two breeds and provide a basis for future research of the molecular mechanisms underlying the piglet differences.
Project description:The cytokine inducible SH2-containing protein (CISH), which might play a role in porcine intestine immune responses, was one of the promising candidate genes for piglet anti-disease traits. An experiment was conducted to characterize the porcine CISH (pCISH) gene and to evaluate its genetic effects on pig anti-disease breeding.Both reverse transcription polymerase chain reaction (RT-PCR) and PCR were performed to obtain the sequence of pCISH gene. A pEGFP-C1-CISH vector was constructed and transfected into PK-15 cells to analysis the distribution of pCISH. The sequences of individuals were compared with each other to find the polymorphisms in pCISH gene. The association analysis was performed in Min pigs and Landrace pigs to evaluate the genetic effects on piglet diarrhea traits.In the present research, the coding sequence and genomic sequence of pCISH gene was obtained. Porcine CISH was mainly localized in cytoplasm. TaqI and HaeIII PCR restriction fragment length polymorphism (RFLP) assays were established to detect single nucleotide polymorphisms (SNPs); A-1575G in promoter region and A2497C in Intron1, respectively. Association studies indicated that SNP A-1575G was significantly associated with diarrhea index of Min piglets (p<0.05) and SNP A2497C was significantly associated with the diarrhea trait of both Min pig and Landrace piglets (p<0.05).This study suggested that the pCISH gene might be a novel candidate gene for pig anti-disease traits, and further studies are needed to confirm the results of this preliminary research.
Project description:The objective of this study is to determine the effect of maternal folate deficiency on the skeletal muscle transcriptome of piglets from a reciprocal cross, in which full-sibling Landrace (LR) and full-sibling Chinese local breed Laiwu (LW) pigs were used for reciprocal cross matings, and sows were fed either a folate deficient or a normal diet during early-mid gestation. In addition, the difference in the responsiveness of the piglets to folate deficiency during early-mid pregnancy between reciprocal cross groups was investigated.Longissimus dorsi (LD) muscle samples were collected from newborn piglets and a 4 x 44K Agilent porcine oligo microarray was used for transcriptome analysis of porcine LD muscle. The results showed that folate deficiency during early-mid pregnancy affected piglet body weight, LD muscle fiber number and content of intramuscular triglyceride.
Project description:Folate deficiency (FD) during pregnancy can cause fetal intrauterine growth restriction in pigs, of which the skeletal dysplasia is a major manifestation. Factors influencing muscle development are very important in the formation of porcine meat quality trait. However, the effect of folate deficiency on skeletal muscle development and its molecular mechanisms are unknown. The objective of this study is to determine the effect of maternal folate deficiency on the skeletal muscle transcriptome of piglets from a reciprocal cross, in which full-sibling Landrace (LR) and full-sibling Chinese local breed Laiwu (LW) pigs were used for reciprocal cross matings, and sows were fed either a folate deficient or a normal diet during early-mid gestation. In addition, the difference in the responsiveness of the piglets to folate deficiency during early-mid pregnancy between reciprocal cross groups was investigated. Longissimus dorsi (LD) muscle samples were collected from newborn piglets and a 4 × 44K Agilent porcine oligo microarray was used for transcriptome analysis of porcine LD muscle. The results showed that folate deficiency during early-mid pregnancy affected piglet body weight, LD muscle fiber number and content of intramuscular triglyceride. The microarray results indicated that 3154 genes were differentially expressed between folate deficient and normal piglets from the LR? × LW? cross, and 3885 differentially expressed genes (DEGs) in the ones from the LW? × LR? cross. From functional analyses, sow folate deficiency affected almost all biological processes in the progeny. Lipid metabolism-related genes and associated metabolic pathways were regulated extensively by folate deficiency, especially in LR? × LW? cross piglets. Most of the genes that are regulated by folate deficiency in the LD muscle of piglets were different between LR? × LW? and LW? × LR? crosses, suggesting some epigenetic effects of FD exist in genes underlying myogenesis and intramuscular fat deposition in piglets.
Project description:Low birth weight (LBWT) is consistently associated with impaired postnatal muscle growth in mammals. Satellite cell (SC)-mediated myonuclear incorporation precedes protein accumulation in the early stages of postnatal muscle development and growth. The objective of this study was to investigate proliferation and differentiation of SCs and the regulation of protein synthesis signaling in response to insulin-like growth factor (IGF)-I stimulation in SC-derived myotubes of LBWT neonatal pigs. SCs isolated from Longissimus dorsi muscle of LBWT and NBWT pigs (3-d-old, n = 8) were cultured and induced to proliferate and differentiate to myotubes in vitro. On day 3 of differentiation, myotubes were fasted in serum-free media for 3 h and treated with human recombinant R3-insulin-like growth factor-I (rh IGF-I) at 0, 25, and 50 ng × mL-1 for 30 min. There was no difference in proliferation rates of SCs from LBWT and NBWT pigs. However, LBWT SC fusion was 15% lower (P ? 0.05) without a difference in MyoD or myogenin mRNA expression in comparison with NBWT pigs, suggesting SCs are not intrinsically different between the two groups. IGF-? stimulation at physiological concentrations activated downstream effectors of mTOR similarly in myotubes from LBWT and NBWT pigs. However, abundance of ribosomal protein S6 kinase 1(S6K1) was lower in myotubes of LBWT compared to their NBWT siblings (P ? 0.05). These results indicate that the modest reduction in SC fusion and S6K1 expression are not the major contributors to the impaired postnatal muscle growth of LBWT pigs.
Project description:This study evaluated the effects of l-carnitine (CAR) and sugar beet pulp (SBP) inclusion in gilt gestation diets on gilt live weight, cortisol concentration, lactation feed intake, and lifetime growth of progeny. Eighty-four pregnant gilts (Large White × Landrace) were randomly assigned to a treatment at day 38 of gestation until parturition; Control (0% SBP, 0 g CAR), CAR (0.125 g/d CAR), SBP (40% SBP), and SBP plus CAR (40% SBP, 0.125 g/d CAR). Gilts were weighed and back-fat depth was recorded on day 38, day 90, and day 108 of gestation and at weaning. Gilt saliva samples were collected pre-farrowing and fecal consistency was scored from entry to the farrowing room until day 5 post-partum. The number of piglets born (total, live, and stillborn) and individual birth weight was recorded. Piglet blood glucose concentration was measured 24 h post-partum and pigs were weighed on day 1, day 6, day 14, day 26, day 76, day 110, and day 147 of life. Carcass data were collected at slaughter. There was no interaction between CAR and SBP for any variable measured. The SBP-fed gilts were heavier on day 90 and day 108 of gestation (P < 0.05) and lost more weight during lactation (P < 0.05) than control gilts. They also had a greater fecal consistency score (P < 0.01). Total farrowing duration, piglet birth interval, and lactation feed intakes were similar between treatments (P > 0.05). The number of piglets born (total, live, and stillborn) and piglet birth weight was likewise similar between treatments (P > 0.05). Piglets from CAR-fed gilts had lower blood glucose concentrations (P < 0.01), while piglets from SBP-fed gilts had greater blood glucose concentrations (P < 0.01). Piglets from CAR gilts had a lower average daily gain between day 1 and day 6 (P < 0.05) and day 14 and day 26 post-partum (P < 0.05) compared to piglets from control gilts. However, CAR gilts weaned a greater number of pigs (P = 0.07). Live weight and carcass weight at slaughter were heavier for pigs from CAR gilts (P < 0.05) and from SBP gilts (P < 0.05). Pigs from CAR gilts (P < 0.01) and SBP gilts (P < 0.05) had increased carcass muscle depth. In conclusion, no benefit was found from the combined feeding of CAR and SBP. Fed separately, CAR increased the live weight, carcass weight, and muscle depth of progeny at slaughter. Feeding a high SBP diet increased fecal consistency in gilts pre-farrowing and increased live weight and carcass muscle depth of progeny.
Project description:Weaning significantly alters hepatic aromatic amino acid (AAA) metabolism and physiological functions. However, less is known about the regulating mechanism of hepatic AAA metabolism after weaning. A total of 200 21-day-old piglets (Duroc × Landrace) were assigned randomly to the control group and the weaning group. In this study, weaning significantly decreased the concentration of phenylalanine, tryptophan, and tyrosine in piglet livers (p < 0.05). Additionally, through the detection of liver AAA metabolites and metabolic enzyme activity, it was observed that hepatic tryptophan catabolism was enhanced, while that of phenylalanine was weakened (p < 0.05). Intriguingly, acetyl-proteome profiling of liver from weaned piglets showed that weaning exacerbated the acetylation of phenylalanine hydroxylase (PAH) and the deacetylation of tryptophan 2,3-dioxygenase (TDO). Analysis of PAH and TDO acetylation in Chang liver cells showed that acetylation decreased the PAH activity, while deacetylation increased the TDO activity (p < 0.05). Furthermore, metabolites of AAAs and the acetylation statuses of PAH and TDO in primary hepatocytes from weaned piglets were consistent with the results in vivo. These findings indicated that weaning altered the PAH and TDO activity by affecting the acetylation state of the enzyme in piglets'' livers. Lysine acetylation may be a potential regulatory mechanism for AAA metabolism in response to weaning.
Project description:The experiment was conducted to study the effect of the glutamate (Glu) on muscle protein loss through toll-like receptor 4 (TLR4), nucleotide-binding oligomerization domain proteins (NODs), Akt/Forkhead Box O (Akt/FOXO) and mammalian target of rapamycin (mTOR) signaling pathways in LPS-challenged piglets. Twenty-four weaned piglets were assigned into four treatments: (1) Control; (2) LPS+0% Glu; (3) LPS + 1.0% Glu; (4) LPS + 2.0% Glu. The experiment was lasted for 28 days. On d 28, the piglets in the LPS challenged groups were injected with LPS on 100 ?g/kg body weight (BW), and the piglets in the control group were injected with the same volume of 0.9% NaCl solution. After 4 h LPS or saline injection, the piglets were slaughtered and the muscle samples were collected. Glu supplementation increased the protein/DNA ratio in gastrocnemius muscle, and the protein content in longissimus dorsi (LD) muscle after LPS challenge (P<0.05). In addition, Glu supplementation decreased TLR4, IL-1 receptor-associated kinase (IRAK) 1, receptor-interacting serine/threonine-protein kinase (RIPK) 2, and nuclear factor-?B (NF-?B) mRNA expression in gastrocnemius muscle (P<0.05), MyD88 mRNA expression in LD muscle, and FOXO1 mRNA expression in LD muscle (P<0.05). Moreover, Glu supplementation increased p-Akt/t-Akt ratio (P<0.05) in gastrocnemius muscle, and p-4EBP1/t-4EBP1 ratio in both gastrocnemius and LD muscles (P<0.05). Glu supplementation in the piglets' diets might be an effective strategy to alleviate LPS-induced muscle protein loss, which might be due to suppressing the mRNA expression of TLR4 and NODs signaling-related genes, and modulating Akt/FOXO and mTOR signaling pathways.
Project description:Maternal care behaviour is crucial for offspring quality and survival in pigs. Defining care is therefore essential for ensuring the welfare of pigs and sustainability of pig production. The aim of the present study was to investigate the relationship between sow nest building, communication with piglets (sniffing, nudging, grunting) during resting and activity, and piglet survival in three different sow breeds: a maternal line selected for high weaned pig production (Landrace), a paternal line selected for meat traits (Duroc), and a crossbred line (Landrace and Yorkshire). We predicted that a higher frequency of nest building and sow communication would have a positive impact on piglet survival. Secondly, we predicted that a high level of maternal care outside the time of nursing (nest building and communication) would increase the quality of the litter (weight at weaning). We also predicted that nest building activity and sow communication would be more pronounced in maternal sow lines selected for maternal traits than in a non-selected, paternal line, and that primiparous sows would perform more nest building behaviour and communicate more than multiparous sows due to high investment in their first litter. Finally, an impaired condition around farrowing (i.e. low body condition score and presence of shoulder lesions) was predicted to be negatively correlated to care behaviours. Data was collected on 38 sows with 511 born piglets. Sows with their litters, were loose-housed in individual farrowing pens until weaning. Nest building activity can be partly considered as maternal care behaviour as it prepares the sows for motherhood and is associated with a lower proportion of stillborn piglets (P < 0.001), starved piglets (P = 0.004), and overlaid piglets (P = 0.034). As predicted, sows that communicated more while being active had lower postnatal piglet mortality (starvation (P < 0.001), less overlying (P < 0.001), overlying with (P < 0.001), and without the milk in the stomach (P < 0.001) and fewer that died of other causes (P < 0.001), higher piglet survival (P < 0.001) and litter weight (P < 0.001) at weaning irrespective of the breed. A higher level of communication while active was associated with more pronounced shoulder lesions in sows (P = 0.010), suggesting a positive association between good maternal care and prevalence of shoulder lesions. We also found that resting sows that communicated more with piglets outside the time of nursing, had higher postnatal piglet mortality (P < 0.001) due to starvation (P < 0.001), overlying (P < 0.001), overlying with (P < 0.001), or without milk (P < 0.001). Communication during resting was more pronounced with increasing litter size at birth (P < 0.001), especially for thin sows (P < 0.001). Communication during resting was more pronounced in the non-selected Duroc line (P < 0.001). Our results suggest that sow communication while being active is a good predictor of good maternal care, piglet survival and litter quality in three different breeds of domestic pigs.