Dietary supplementation of laminarin improves the reproductive performance of sows and the growth of suckling piglets

Background Maternal nutrition is essential in keeping a highly efficient production system in the pig industry.Laminarin has been shown to improve antioxidant capacity,reduce the inflammatory response,and favor the homeostasis of intestinal microbiota.However,the effect of dietary supplementation of laminarin on the reproductive performance of sows and the growth of suckling offspring remains unknown.Methods A total of 40 Landrace×Yorkshire multiparous sows on d 85 of gestation,similar in age,body weight(BW),parity and reproductive performance,were randomly divided into four dietary treatments with 10 sows per treatment,receiving a control diet(basal pregnancy or lactating diets)and a basal diet supplemented with 0.025%,0.05%and 0.10%laminarin,respectively.The experiment lasted from d 85 of gestation to d 21 of lactation.Results Laminarin supplementation linearly increased number born alive per litter(P=0.03),average daily feed intake(ADFI,P<0.01),and total milk yield of sows during the lactation of 1–21 d(P=0.02).Furthermore,maternal laminarin supplementation increased the average daily gain(ADG)of piglets while tending to reduce the culling and death rate before weaning.In addition,alterations to the composition of colostrum and milk,as well as to serum inflammatory cytokines and immunoglobulins of sows were observed.The fecal microbiota profile of sows supported the improvement of reproductive performance in sows and the growth performance in suckling offspring.Conclusions Dietary supplementation of laminarin during late pregnancy and lactation could significantly improve reproductive performance of sows and growth performance of piglets.

Global transcriptomic analysis reveals Lnc-ADAMTS9 exerting an essential role in myogenesis through modulating the ERK signaling pathway

Background:Long non-coding RNAs(lncRNAs)are emerging key regulators involved in a variety of biological processes such as cell differentiation and development.The balance between myogenesis and adipogenesis is crucial for skeletal muscle homeostasis in humans and meat quality in farm animals.The present study aimed to reveal the global transcriptomic profiles of adipogenic(Adi-)and myogenic(Myo-)precursors derived from porcine skeletal muscle and identify lncRNAs involved in the modulation of myogenesis homeostasis in porcine skeletal muscle.Results:In this study,a total of 655 novel individual lncRNAs including differentially expressed 24 lncRNAs,and 755 differentially expressed mRNAs were identified(fold change≥2 or≤0.5 and adjusted P<0.05).Integrated results of Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analysis accompanied by the variation of intracellular Ca2+concentration highlighted Lnc-ADAMTS9 involved in the modulation of myogenesis homeostasis in porcine skeletal muscle.Although Lnc-ADAMTS9 knock-down did not alter the mRNA expression of ADAMTS9,we demonstrated that Lnc-ADAMTS9 can promote myogenic proliferation and myogenic differentiation of myogenic precursors through inhibiting the ERK/MAPK signaling pathway.Conclusion:We deciphered a comprehensive catalog of mRNAs and lncRNAs that might be involved in the regulation of myogenesis and adipogenesis homeostasis in the skeletal muscle of pigs.The Lnc-ADAMTS9 exerts an essential role in myogenesis through the ERK signaling pathway.

Ryanodine receptor RyR1-mediated elevation of Ca^(2+)concentration is required for the late stage of myogenic differentiation and fusion

Background:Cytosolic Ca^(2+)plays vital roles in myogenesis and muscle development.As a major Ca^(2+)release channel of endoplasmic reticulum(ER),ryanodine receptor 1(RyR1)key mutations are main causes of severe congenital myopathies.The role of RyR1 in myogenic differentiation has attracted intense research interest but remains unclear.Results:In the present study,both RyR1-knockdown myoblasts and CRISPR/Cas9-based RyR1-knockout myoblasts were employed to explore the role of RyR1 in myogenic differentiation,myotube formation as well as the potential mechanism of RyR1-related myopathies.We observed that RyR1 expression was dramatically increased during the late stage of myogenic differentiation,accompanied by significantly elevated cytoplasmic Ca^(2+)concentration.Inhibition of RyR1 by siRNA-mediated knockdown or chemical inhibitor,dantrolene,significantly reduced cytosolic Ca^(2+)and blocked multinucleated myotube formation.The elevation of cytoplasmic Ca^(2+)concentration can effectively relieve myogenic differentiation stagnation by RyR1 inhibition,demonstrating that RyR1 modulates myogenic differentiation via regulation of Ca^(2+)release channel.However,RyR1-knockout-induced Ca^(2+)leakage led to the severe ER stress and excessive unfolded protein response,and drove myoblasts into apoptosis.Conclusions:Therefore,we concluded that Ca^(2+)release mediated by dramatic increase in RyR1 expression is required for the late stage of myogenic differentiation and fusion.This study contributes to a novel understanding of the role of RyR1 in myogenic differentiation and related congenital myopathies,and provides a potential target for regulation of muscle characteristics and meat quality.

Lysine 2-hydroxyisobutyrylation levels determined adipogenesis and fat accumulation in adipose tissue in pigs

Background Excessive backfat deposition lowering carcass grade is a major concern in the pig industry,especially in most breeds of obese type pigs.The mechanisms involved in adipogenesis and fat accumulation in pigs remain unclear.Lysine 2-hydroxyisobutyrylation(Khib),is a novel protein post-translational modification(PTM),which play an important role in transcription,energy metabolism and metastasis of cancer cells,but its role in adipogenesis and fat accumulation has not been shown.Results In this study,we first analyzed the modification levels of acetylation(Kac),Khib,crotonylation(Kcr)and succinylation(Ksu)of fibro-adipogenic progenitors(FAPs),myogenic precursors(Myo)and mesenchymal stem cells(MSCs)with varied differentiation potential,and found that only Khib modification in FAPs was significantly higher than that in MSCs.Consistently,in parallel with its regulatory enzymes lysine acetyltransferase 5(KAT5)and histone deacetylase 2(HDAC2)protein levels,the Khib levels increased quadratically(P<0.01)during adipogenic differentiation of FAPs.KAT5 knockdown in FAPs inhibited adipogenic differentiation,while HDAC2 knockdown enhanced adipogenic differentiation.We also demonstrated that Khib modification favored to adipogenic differentiation and fat accumulation by comparing Khib levels in FAPs and backfat tissues both derived from obese-type pigs(Laiwu pigs)and lean-type pigs(Duroc pigs),respectively.Accordingly,the expression patterns of KAT5 and HDAC2 matched well to the degree of backfat accumulation in obese-and lean-type pigs.Conclusions From the perspective of protein translational modification,we are the first to reveal the role of Khib in adipogenesis and fat deposition in pigs,and provided new clues for the improvement of fat accumulation and distribution as expected via genetic selection and nutritional strategy in obese-type pigs.

Arginine promotes myogenic differentiation and myotube formation through the elevation of cytoplasmic calcium concentration

This study aimed to explore the mechanism underlying arginine-promoted myogenesis of myoblasts.C2C12 cells were cultured with a medium containing 0.1,0.4,0.8,or 1.2 mmol/L arginine,respectively.Cell proliferation,viability,differentiation indexes,cytoplasmic Ca^(2+)concentration,and relative mRNA expression levels of myogenic regulatory factors(MRF)and key Ca2+channels were measured in the absence or presence of 2 chemical inhibitors,dantrolene(DAN,10μmol/L)and nisoldipine(NIS,10μmol/L),respectively.Results demonstrated that arginine promoted myogenic differentiation and myotube formation.Compared with the control(0.4 mmol/L arginine),1.2 mmol/L arginine upregulated the relative mRNA expression levels of myogenin(MyoG)and Myomaker at d 2 during myogenic induction(P<0.05).Cytoplasmic Ca^(2+)concentrations were significantly elevated by arginine supplementation at d 2 and 4(P<0.05).Relative mRNA expression levels of Ca^(2+)channels including the type 1 ryanodine recepto r(RyR1)and voltage-gated Ca^(2+)channel(Cav1.1)were upregulated by 1.2 mmol/L arginine during2-d myogenic induction(P<0.01).However,arginine-promoted myogenic potential of myoblasts was remarkably compromised by DAN and NIS,respectively(P<0.05).These findings evidenced that the supplementation of arginine promoted myogenic differentiation and myotube formation through increasing cytoplasmic Ca^(2+)concentration from both extracellular and sarcoplasmic reticulum Ca^(2+).

Dietary guanidinoacetic acid supplementation improves water holding capacity and lowers free amino acid concentration of fresh meat in finishing pigs fed with various dietary protein levels

The current study was carried out to detect the effect of dietary guanidinoacetic acid(GAA)supplementation on carcass characteristics and meat quality in finishing pigs fed different dietary crude protein(CP)levels.Sixty-four barrows with an initial body weight of 73.05±2.34 kg were randomly allocated into 1 of 4 dietary treatments in a 2(100%vs.125%NRC CP level)×2(0 vs.300 mg/kg GAA)factorial arrangement(n=7).The feeding trial lasted for 49 d.GAA supplementation significantly reduced drip loss(P=0.01),free water distribution(T23peak area ratio)(P=0.05)and the concentrations of free alanine,threonine,methionine and isoleucine(P<0.05);but increased total glycine content(P=0.03)in the longissimus dorsi muscle of finishing pigs regardless of the dietary CP levels.Furthermore,primary myogenic cell differentiation system was employed to investigate the influence of inclusion of GAA on free amino acid concentrations in myotubes(n=4)and validate the finding in the animal feeding trial.We found that GAA inclusion in culture medium also decreased intracellular concentrations of free alanine,threonine,methionine,isoleucine,valine and proline in differentiated primary myogenic cells in vitro(P<0.05).Meanwhile,relative to diets with 100%NRC CP level,the intake of diets with 125%NRC CP level improved sarcoplasmic protein solubility,increased the contents of carnosine and total free amino acids as well as flavor amino acids in the longissimus dorsi muscle and decreased backfat thickness at the 6-7th ribs in pigs(P<0.05).In addition,we observed that the impact of dietary GAA supplementation on the last rib fat thickness,shear force,and free lysine content in the longissimus dorsi muscle was dependent on dietary CP levels(P<0.05).Collectively,dietary GAA supplementation can reduce drip loss,decrease the concentrations of free amino acids and flavor amino acids of fresh meat independent of dietary CP levels.