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.

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.

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.

Nutritional regulation of skeletal muscle energy metabolism, lipid accumulation and meat quality in pigs

The quality of pork determines consumers’ purchase intention, which directly affects the economic value of pork. Minimizing the proportion of inferior pork and producing high quality pork are the ultimate goals of the pig industry. Muscle energy metabolism, serving as a regulative hub in organism energy expenditure and storage as a fat deposit, is compatible with myofiber type composition, affecting meat color, intramuscular fat content, tenderness, p H values and drip loss. Increasing data illustrate that dietary nutrients and bioactive ingredients affect muscle energy metabolism, white adipose browning and fat distribution, and myofiber type composition in humans, and rodents. Recently, some studies have shown that modulating muscle energy metabolism and lipid accumulation through nutritional approaches could effectively improve meat quality. This article reviews the progress and development in this field, and specifically discusses the impacts of dietary supply of amino acids, lipids, and gut microbiota as well as maternal nutrition on skeletal muscle energy metabolism, lipid accumulation and meat quality of pigs, so as to provide comprehensive overview with respect to effective avenues for improving meat quality.