Background Undernutrition disrupts pregnant ewe's metabolic homeostasis and severely inhibits fetal growth and development.In this study,undernourished and nutrition-recovery pregnant sheep models and rumen epithe...Background Undernutrition disrupts pregnant ewe's metabolic homeostasis and severely inhibits fetal growth and development.In this study,undernourished and nutrition-recovery pregnant sheep models and rumen epithelial cells were utilized to investigate the mechanisms behind undernutrition-induced disruptions in male fetal rumen metabolism and development.Results Maternal undernutrition significantly reduced male fetal rumen weight and papilla length,width and surface area.Maternal undernutrition extremely suppressed nutrient metabolism and energy production in male fetal rumen via JAK3/STAT3 signaling to inhibit cell cycle progression and male fetal rumen development,while maternal nutritional recovery partially restored metabolic inhibition but failed to alleviate male fetal rumen development.Meanwhile,64 differentially expressed miRNAs(DEMs)were identified in male fetal rumen between undernourished ewes and controls.Novel miR-736 was overexpressed both in male fetal rumen of undernourished and nutrition-recovery models.E2F transcription factor 2(E2F2)and MYB proto-oncogene like 2(MYBL2)were the intersection of male fetal rumen differentially expressed genes(DEGs)and DEMs target genes integrated analysis and were predicted as novel miR-736 target genes.Further,we confirmed that novel miR-736 targeted and downregulated E2F2 and MYBL2 expression levels.Silencing E2F2 and MYBL2 promoted apoptosis and inhibited S-phase entry in rumen epithelial cells.Conclusions In summary,maternal undernutrition disrupted male fetal rumen metabolism and elevated novel miR-736,which targeted and downregulated E2F2 and MYBL2 to inhibit cell cycle progression and promote apoptosis,finally inhibited male fetal rumen development.This study provides new insights into the epigenetic mechanisms underlying maternal undernutrition-induced male fetal rumen developmental deficits.展开更多
Background:This study aimed to elucidate the molecular mechanisms of solid diet introduction initiating the cellular growth and maturation of rumen tissues and characterize the shared and unique biological processes u...Background:This study aimed to elucidate the molecular mechanisms of solid diet introduction initiating the cellular growth and maturation of rumen tissues and characterize the shared and unique biological processes upon different solid diet regimes.Methods:Twenty-four Hu lambs were randomly allocated to three groups fed following diets:goat milk powder only(M,n=8),goat milk powder+alfalfa hay(MH,n=8),and goat milk powder+concentrate starter(MC,n=8).At 42 days of age,the lambs were slaughtered.Ruminal fluid sample was collected for analysis of concentration of volatile fatty acid(VFA)and microbial crude protein(MCP).The sample of the rumen wall from the ventral sac was collected for analysis of rumen papilla morphology and transcriptomics.Results:Compared with the M group,MH and MC group had a higher concentration of VFA,MCP,rumen weight,and rumen papilla area.The transcriptomic results of rumen wall showed that there were 312 shared differentially expressed genes(DEGs)between in“MH vs.M”and“MC vs.M”,and 232 or 796 unique DEGs observed in“MH vs.M”or“MC vs.M”,respectively.The shared DEGs were most enriched in VFA absorption and metabolism,such as peroxisome proliferator-activated receptor(PPAR)signaling pathway,butanoate metabolism,and synthesis and degradation of ketone bodies.Additionally,a weighted gene co-expression network analysis identified M16(2,052 genes)and M18(579 genes)modules were positively correlated with VFA and rumen wall morphology.The M16 module was mainly related to metabolism pathway,while the M18 module was mainly associated with signaling transport.Moreover,hay specifically depressed expression of genes involved in cytokine production,immune response,and immunocyte activation,and concentrate starter mainly altered nutrient transport and metabolism,especially ion transport,amino acid,and fatty acid metabolism.Conclusions:The energy production during VFA metabolism may drive the rumen wall development directly.The hay introduction facilitated establishment of immune function,while the concentrate starter enhanced nutrient transport and metabolism,which are important biological processes required for rumen development.展开更多
The role ofβ-hydroxybutyric acid(BHBA)includes providing energy,regulating signaling pathways,and ameliorating the gut microbiota in the host,while its nutrient mechanism to improve rumen epithelium development in yo...The role ofβ-hydroxybutyric acid(BHBA)includes providing energy,regulating signaling pathways,and ameliorating the gut microbiota in the host,while its nutrient mechanism to improve rumen epithelium development in young ruminants is still unclear.In this study,a total of 12 female Haimen goats with 30 d of age were chosen and divided into two groups.One group was fed with basic diet(CON),and the other group was fed a basal diet supplemented with 6 g d^(-1)dietaryβ-hydroxybutyrate sodium(BHBA-Na).The experimental period was 30 d,and all goats were slaughtered at 60 d of age.The joint analysis of multi-omics,including rumen microbiota,rumen epithelial transcriptome and rumen epithelial metabolomics in young goat model,was performed to systematically investigate the effect of dietary BHBA-Na on rumen development in young goats.As the results,we found that dietary BHBA-Na improved the growth performance of young goat including body weight,average daily gain(ADG)and dry matter intake(DMI)(P<0.05).Dietary BHBA-Na also increased the weight of rumen,and promoted the growth of rumen epithelium development(P<0.05).The abundance of several beneficial bacteria was increased(Fibrobacter,Succinivibrio,Clostridiales,etc.).The rumen epithelium transcriptome and metabolomics indicated that BHBA-Na supplementation showed a remarkable effect on the nutrient metabolism of the rumen epithelium.Specifically,the pathways of“fatty acid metabolism”,“cholesterol homeostasis”,“reactive oxygen species(ROS)pathway”and“peroxisome”were activated in response to BHBA-Na addition(P<0.05).Moreover,the genes(HMGCS2,ECSH1,ACAA2,ECH1,ACADS etc.)and metabolites(succinic acid,alpha-ketoisovaleric acid,etc.)involved in these pathways were also regulated positively(P<0.05).The rumen epithelium obtained the energy for its development from the process of volatile fatty acids(VFAs)decomposition.Finally,we observed the close correlations among the phenotypes,ruminal microbiota,host genes and epithelial metabolites.Overall,our results revealed that the BHBA-Na promoted the growth and rumen development of young goats possibly by enhancing DMI and regulating the rumen microbiota and the metabolisms of VFA and amino acid in the rumen epithelium.展开更多
The rumen is an important organ that enables ruminants to digest nutrients. However, the biological mechanism by which the microbiota and its derived fatty acids regulate rumen development is still unclear. In this st...The rumen is an important organ that enables ruminants to digest nutrients. However, the biological mechanism by which the microbiota and its derived fatty acids regulate rumen development is still unclear. In this study, 18 female Haimen goats were selected and slaughtered at d 30, 60, and 90 of age.Multi-omics analyses(rumen microbial sequencing, host transcriptome sequencing, and rumen epithelial metabolomics) were performed to investigate host-microbe interactions from preweaning to postweaning in a goat model. With increasing age, and after the introduction of solid feed, the increased abundances of Prevotella and Roseburia showed positive correlations with volatile fatty acid(VFA) levels and morphological parameters(P < 0.05). Epithelial transcriptomic analysis showed that the expression levels of hub genes, including 3-hydroxy-3-methylglutaryl-CoA synthase isoform 2(HMGCS2), enoyl-CoA hydratase, short chain 1(ECHS1), and peroxisome proliferator activated receptor gamma(PPARG), were positively associated with animal phenotype(P < 0.05). These hub genes were mainly correlated to VFA metabolism, oxidative phosphorylation, and the mammalian target of rapamycin(mTOR) and peroxisome proliferator activated receptor(PPAR) signaling pathways(P < 0.05). Moreover, the primary metabolites in the epithelium changed from glucose preweaning to(R)-3-hydroxybutyric acid(BHBA) and acetoacetic acid(ACAC) postweaning(P < 0.05). Diet and butyrate were the major factors shaping epithelial metabolomics in young ruminants(P < 0.05). Multi-omics analysis showed that the rumen microbiota and VFA were mainly associated with the epithelial transcriptome, and that alterations in gene expression influenced host metabolism. The “butanoate metabolism” pathway, which transcriptomic and metabolomic analyses identified as being upregulated with age, produces ketones that regulate the “oxidative phosphorylation” pathway, which could provide energy for the development of rumen papillae. Our findings reveal the changes that occur in the rumen microbiota, host transcriptome,and metabolome with age, and validate the role of microbiota-derived VFA in manipulating host gene expression and subsequent metabolism. This study provides insight into the molecular mechanisms of host-microbe interactions in goats and supplies a theoretical basis and guidance for precise nutritional regulation during the critical time window for rumen development of young ruminants.展开更多
基金supported by the National Natural Science Foundation of China(32402767)National Key Research and Development Program of China(2022YFD1301102)+3 种基金Anhui Province Natural Science Foundation Youth Project(2308085QC104)AAU Introduction of High-level Talent Funds(RC392107)Key Laboratory of Utilization of Livestock and Forage Resources in Circum-Tarim Region(Co-construction by Ministry and Province),Ministry of Agriculture and Rural Affairs(BSGJSYS202502)the China Agriculture Research System(CARS-38)。
文摘Background Undernutrition disrupts pregnant ewe's metabolic homeostasis and severely inhibits fetal growth and development.In this study,undernourished and nutrition-recovery pregnant sheep models and rumen epithelial cells were utilized to investigate the mechanisms behind undernutrition-induced disruptions in male fetal rumen metabolism and development.Results Maternal undernutrition significantly reduced male fetal rumen weight and papilla length,width and surface area.Maternal undernutrition extremely suppressed nutrient metabolism and energy production in male fetal rumen via JAK3/STAT3 signaling to inhibit cell cycle progression and male fetal rumen development,while maternal nutritional recovery partially restored metabolic inhibition but failed to alleviate male fetal rumen development.Meanwhile,64 differentially expressed miRNAs(DEMs)were identified in male fetal rumen between undernourished ewes and controls.Novel miR-736 was overexpressed both in male fetal rumen of undernourished and nutrition-recovery models.E2F transcription factor 2(E2F2)and MYB proto-oncogene like 2(MYBL2)were the intersection of male fetal rumen differentially expressed genes(DEGs)and DEMs target genes integrated analysis and were predicted as novel miR-736 target genes.Further,we confirmed that novel miR-736 targeted and downregulated E2F2 and MYBL2 expression levels.Silencing E2F2 and MYBL2 promoted apoptosis and inhibited S-phase entry in rumen epithelial cells.Conclusions In summary,maternal undernutrition disrupted male fetal rumen metabolism and elevated novel miR-736,which targeted and downregulated E2F2 and MYBL2 to inhibit cell cycle progression and promote apoptosis,finally inhibited male fetal rumen development.This study provides new insights into the epigenetic mechanisms underlying maternal undernutrition-induced male fetal rumen developmental deficits.
基金This work was supported by the Project for Top Young Talents Program of College of Animal Science and Technology of Nanjing Agricultural University(DKQB201904)National Key Research and Development Plan(2018YFD0501900)+1 种基金Postgraduate Research&Practice Innovation Program of Jiangsu Province(KYCX20_0603)Science and Technology Project of Huzhou City of China(2017GY18).
文摘Background:This study aimed to elucidate the molecular mechanisms of solid diet introduction initiating the cellular growth and maturation of rumen tissues and characterize the shared and unique biological processes upon different solid diet regimes.Methods:Twenty-four Hu lambs were randomly allocated to three groups fed following diets:goat milk powder only(M,n=8),goat milk powder+alfalfa hay(MH,n=8),and goat milk powder+concentrate starter(MC,n=8).At 42 days of age,the lambs were slaughtered.Ruminal fluid sample was collected for analysis of concentration of volatile fatty acid(VFA)and microbial crude protein(MCP).The sample of the rumen wall from the ventral sac was collected for analysis of rumen papilla morphology and transcriptomics.Results:Compared with the M group,MH and MC group had a higher concentration of VFA,MCP,rumen weight,and rumen papilla area.The transcriptomic results of rumen wall showed that there were 312 shared differentially expressed genes(DEGs)between in“MH vs.M”and“MC vs.M”,and 232 or 796 unique DEGs observed in“MH vs.M”or“MC vs.M”,respectively.The shared DEGs were most enriched in VFA absorption and metabolism,such as peroxisome proliferator-activated receptor(PPAR)signaling pathway,butanoate metabolism,and synthesis and degradation of ketone bodies.Additionally,a weighted gene co-expression network analysis identified M16(2,052 genes)and M18(579 genes)modules were positively correlated with VFA and rumen wall morphology.The M16 module was mainly related to metabolism pathway,while the M18 module was mainly associated with signaling transport.Moreover,hay specifically depressed expression of genes involved in cytokine production,immune response,and immunocyte activation,and concentrate starter mainly altered nutrient transport and metabolism,especially ion transport,amino acid,and fatty acid metabolism.Conclusions:The energy production during VFA metabolism may drive the rumen wall development directly.The hay introduction facilitated establishment of immune function,while the concentrate starter enhanced nutrient transport and metabolism,which are important biological processes required for rumen development.
基金funded by the Guangdong Provincial Modern Agricultural Industrial Technology System Innovation Team Construction Project(2024CXTD13)the Key Research and Development Plan Project of the Tibet Autonomous Region(XZ202301ZY0008N)funded by the Regional Science and Technology Collaborative Innovation Project of the Tibet Autonomous Region(SNQYKJXT-03 and QYXTZX-RKZ2024-03-1)。
文摘The role ofβ-hydroxybutyric acid(BHBA)includes providing energy,regulating signaling pathways,and ameliorating the gut microbiota in the host,while its nutrient mechanism to improve rumen epithelium development in young ruminants is still unclear.In this study,a total of 12 female Haimen goats with 30 d of age were chosen and divided into two groups.One group was fed with basic diet(CON),and the other group was fed a basal diet supplemented with 6 g d^(-1)dietaryβ-hydroxybutyrate sodium(BHBA-Na).The experimental period was 30 d,and all goats were slaughtered at 60 d of age.The joint analysis of multi-omics,including rumen microbiota,rumen epithelial transcriptome and rumen epithelial metabolomics in young goat model,was performed to systematically investigate the effect of dietary BHBA-Na on rumen development in young goats.As the results,we found that dietary BHBA-Na improved the growth performance of young goat including body weight,average daily gain(ADG)and dry matter intake(DMI)(P<0.05).Dietary BHBA-Na also increased the weight of rumen,and promoted the growth of rumen epithelium development(P<0.05).The abundance of several beneficial bacteria was increased(Fibrobacter,Succinivibrio,Clostridiales,etc.).The rumen epithelium transcriptome and metabolomics indicated that BHBA-Na supplementation showed a remarkable effect on the nutrient metabolism of the rumen epithelium.Specifically,the pathways of“fatty acid metabolism”,“cholesterol homeostasis”,“reactive oxygen species(ROS)pathway”and“peroxisome”were activated in response to BHBA-Na addition(P<0.05).Moreover,the genes(HMGCS2,ECSH1,ACAA2,ECH1,ACADS etc.)and metabolites(succinic acid,alpha-ketoisovaleric acid,etc.)involved in these pathways were also regulated positively(P<0.05).The rumen epithelium obtained the energy for its development from the process of volatile fatty acids(VFAs)decomposition.Finally,we observed the close correlations among the phenotypes,ruminal microbiota,host genes and epithelial metabolites.Overall,our results revealed that the BHBA-Na promoted the growth and rumen development of young goats possibly by enhancing DMI and regulating the rumen microbiota and the metabolisms of VFA and amino acid in the rumen epithelium.
基金funded by grants from National Natural Science Foundation of China(31872385)Foshan Postdoctoral Sustentation Fund(BKS209151)+1 种基金the Inner Mongolia Science and Technology Key Project(2021SZD0014)the National Key R&D Program Projects(2018YFD0501902)。
文摘The rumen is an important organ that enables ruminants to digest nutrients. However, the biological mechanism by which the microbiota and its derived fatty acids regulate rumen development is still unclear. In this study, 18 female Haimen goats were selected and slaughtered at d 30, 60, and 90 of age.Multi-omics analyses(rumen microbial sequencing, host transcriptome sequencing, and rumen epithelial metabolomics) were performed to investigate host-microbe interactions from preweaning to postweaning in a goat model. With increasing age, and after the introduction of solid feed, the increased abundances of Prevotella and Roseburia showed positive correlations with volatile fatty acid(VFA) levels and morphological parameters(P < 0.05). Epithelial transcriptomic analysis showed that the expression levels of hub genes, including 3-hydroxy-3-methylglutaryl-CoA synthase isoform 2(HMGCS2), enoyl-CoA hydratase, short chain 1(ECHS1), and peroxisome proliferator activated receptor gamma(PPARG), were positively associated with animal phenotype(P < 0.05). These hub genes were mainly correlated to VFA metabolism, oxidative phosphorylation, and the mammalian target of rapamycin(mTOR) and peroxisome proliferator activated receptor(PPAR) signaling pathways(P < 0.05). Moreover, the primary metabolites in the epithelium changed from glucose preweaning to(R)-3-hydroxybutyric acid(BHBA) and acetoacetic acid(ACAC) postweaning(P < 0.05). Diet and butyrate were the major factors shaping epithelial metabolomics in young ruminants(P < 0.05). Multi-omics analysis showed that the rumen microbiota and VFA were mainly associated with the epithelial transcriptome, and that alterations in gene expression influenced host metabolism. The “butanoate metabolism” pathway, which transcriptomic and metabolomic analyses identified as being upregulated with age, produces ketones that regulate the “oxidative phosphorylation” pathway, which could provide energy for the development of rumen papillae. Our findings reveal the changes that occur in the rumen microbiota, host transcriptome,and metabolome with age, and validate the role of microbiota-derived VFA in manipulating host gene expression and subsequent metabolism. This study provides insight into the molecular mechanisms of host-microbe interactions in goats and supplies a theoretical basis and guidance for precise nutritional regulation during the critical time window for rumen development of young ruminants.
