Chinese forest musk deer(FMD),an endangered species,have exhibited low reproductive rates even in captivity due to stress conditions.Investigation revealed the presence of di(2-ethylhexyl)phthalate(DEHP),an environmen...Chinese forest musk deer(FMD),an endangered species,have exhibited low reproductive rates even in captivity due to stress conditions.Investigation revealed the presence of di(2-ethylhexyl)phthalate(DEHP),an environmental endocrine disruptor,in the serum and skin of captive FMDs.Feeding FMDs with maslinic acid(MA)has been observed to alleviate the stress response and improve reproductive rates,although the precise molecular mechanisms remain unclear.Therefore,this study aims to investigate the molecular mechanisms underlying the alleviation of DEHP-induced oxidative stress and cell apoptosis in primary peritubular myoid cells(PMCs)through MA intake.Primary PMCs were isolated and exposed to DEHP in vitro.The results demonstrated that DEHP significantly suppressed antioxidant levels and promoted cell apoptosis in primary PMCs.Moreover,interfering with the expression of PRDX6 was found to induce excessive reactive oxygen species(ROS)production and cell apoptosis in primary PMCs.Supplementation with MA significantly upregulated the expression of PRDX6,thereby attenuating DEHP-induced oxidative stress and cell apoptosis in primary PMCs.These findings provide a theoretical foundation for mitigating stress levels and enhancing reproductive capacity of in captive FMDs.展开更多
Background The musk glands of adult male Chinese forest musk deer(Moschus berezovskii Flerov,1929)(FMD),which are considered as special skin glands,secrete a mixture of sebum,lipids,and proteins into the musk pod.Toge...Background The musk glands of adult male Chinese forest musk deer(Moschus berezovskii Flerov,1929)(FMD),which are considered as special skin glands,secrete a mixture of sebum,lipids,and proteins into the musk pod.Together,these components form musk,which plays an important role in attracting females during the breeding season.However,the relationship between the musk glands and skin of Chinese FMD remains undiscovered.Here,the musk gland and skin of Chinese FMD were examined using histological analysis and RNA sequencing(RNA-seq),and the expression of key regulatory genes was evaluated to determine whether the musk gland is derived from the skin.Methods A comparative analysis of musk gland anatomy between juvenile and adult Chinese FMD was conducted.Then,based on the anatomical structure of the musk gland,skin tissues from the abdomen and back as well as musk gland tissues were obtained from three juvenile FMD.These tissues were used for RNA-seq,hematoxylin-eosin(HE)staining,immunohistochemistry(IHC),western blot(WB),and quantitative real-time polymerase chain reaction(qRT-PCR)experiments.Results Anatomical analysis showed that only adult male FMD had a complete glandular organ and musk pod,while juvenile FMD did not have any well-developed musk pods.Transcriptomic data revealed that 88.24%of genes were co-expressed in the skin and musk gland tissues.Kyoto Encyclopedia of Genes and Genomes(KEGG)signaling pathway analysis found that the genes co-expressed in the abdomen skin,back skin,and musk gland were enriched in biological development,endocrine system,lipid metabolism,and other pathways.Gene Ontology(GO)enrichment analysis indicated that the genes expressed in these tissues were enriched in biological processes such as multicellular development and cell division.Moreover,the Metascape predictive analysis tool demonstrated that genes expressed in musk glands were skin tissue-specific.qRT-PCR and WB revealed that sex-determining region Y-box protein 9(Sox9),Caveolin-1(Cav-1),and androgen receptor(AR)were expressed in all three tissues,although the expression levels differed among the tissues.According to the IHC results,Sox9 and AR were expressed in the nuclei of sebaceous gland,hair follicle,and musk gland cells,whereas Cav-1 was expressed in the cell membrane.Conclusions The musk gland of Chinese FMD may be a derivative of skin tissue,and Sox9,Cav-1,and AR may play significant roles in musk gland development.展开更多
Analysis of the intestinal microbiota and physiological parameters in mammalian infancy can reveal health status.In this study,we used a combination of molecular and immunochemical approaches to assess fecal microbiot...Analysis of the intestinal microbiota and physiological parameters in mammalian infancy can reveal health status.In this study,we used a combination of molecular and immunochemical approaches to assess fecal microbiota as well as Cortisol(Cor),Triiodothyronine(T3),and immunoglobulin A(IgA)levels of young forest musk deer(FMD),from birth to one month after weaning(7 days of age–110 days of age).During development as the diet of FMD changes from consuming milk to eating plants,the richness and diversity of intestinal microbiota of young FMD increased significantly.Cor levels remained unchanged throughout early development while significantly increased after weaning,T3 and IgA initially were derived from milk during lactation,significantly decreased after weaning.Correlation network analysis showed that the community of food-oriented microbes were highly structured and that many genera were correlated.Overall,this study provides scientific insights into effective management strategies for the protection of FMD population.展开更多
The forest musk deer(Moschus berezovskii) is one of the most endangered species in China.Over the past decades,extensive hunting and poaching have pushed the forest musk deer to the edge of extinction,and conservation...The forest musk deer(Moschus berezovskii) is one of the most endangered species in China.Over the past decades,extensive hunting and poaching have pushed the forest musk deer to the edge of extinction,and conservation biologists are presently pursuing scientific management plans to rescue this species.The major histocompatibility complex(MHC),a cluster of genes responsible for antigen presentation,is a highly polymorphic genomic region in vertebrates that has become a popular functional marker system for studying adaptive variation.In this study,we developed locus-specific genotyping primers for exon 2 fragments of one DRA gene and one DRB locus of the forest musk deer using a suite of comprehensive methods that included universal primer amplification,genome walking,single-strand conformation polymorphism(SSCP),heteroduplex(HD) profiling,and sequence analysis.Each forest musk deer showed no more than two sequences per locus,confirming the specificity of our primers.Genotyping with these primers allowed us to identify two DRA alleles and six DRB alleles in a captive breeding population of the Sichuan Musk Deer Breeding Institution.For the DRA locus,we found a slightly higher observed heterozygosity(N O =0.154) than expected(N E =0.143).In contrast,the DRB locus showed a significant heterozygote deficiency(N O =0.508;N E =0.761;P<0.05),which was probably due to inbreeding in the captive population.An obvious excess of nonsynonymous substitutions over synonymous was observed at the antigen-binding positions of the DRA and DRB loci,showing the presence of positive selection in the forest musk deer DR genes.Finally,generation of phylogenetic trees for the DRA and DRB sequences of the forest musk deer and other ruminants revealed that the DRA and DRB loci identified in this study had homologous relationships with the known ruminant DRA and DRB genes.Based on this analysis,and to facilitate future studies,we named these novel loci Mobe-DRA and Mobe-DRB3.展开更多
基金supported by the Fund of Sci-Tech Innovation Program of Shaanxi Academy of Forestry(No.SXLK2021-0219)the Science and Technology Project of Shaanxi Province(No.2022SF-512)the Science and Technology Innovation and Achievement Transformation Project of Experimental Demonstration Station(base)of Northwest A&F University(No.TGZX2021-32)。
文摘Chinese forest musk deer(FMD),an endangered species,have exhibited low reproductive rates even in captivity due to stress conditions.Investigation revealed the presence of di(2-ethylhexyl)phthalate(DEHP),an environmental endocrine disruptor,in the serum and skin of captive FMDs.Feeding FMDs with maslinic acid(MA)has been observed to alleviate the stress response and improve reproductive rates,although the precise molecular mechanisms remain unclear.Therefore,this study aims to investigate the molecular mechanisms underlying the alleviation of DEHP-induced oxidative stress and cell apoptosis in primary peritubular myoid cells(PMCs)through MA intake.Primary PMCs were isolated and exposed to DEHP in vitro.The results demonstrated that DEHP significantly suppressed antioxidant levels and promoted cell apoptosis in primary PMCs.Moreover,interfering with the expression of PRDX6 was found to induce excessive reactive oxygen species(ROS)production and cell apoptosis in primary PMCs.Supplementation with MA significantly upregulated the expression of PRDX6,thereby attenuating DEHP-induced oxidative stress and cell apoptosis in primary PMCs.These findings provide a theoretical foundation for mitigating stress levels and enhancing reproductive capacity of in captive FMDs.
基金This work was supported by the Key Sci-Tech Project of Shaanxi Province(No.2018ZDXM-NY-034)the Innovation Capability Support Program of Shaanxi Province(No.2018-TD-021),Chinasupport by the Northwest A&F University and the reproductive biology and cell engineering team.We acknowledge Mr.Xuezhe ZHANG at the Shaanxi Baosen Musk Deer Industry Co.,Ltd.(Shaanxi,China)for providing experimental animals.
文摘Background The musk glands of adult male Chinese forest musk deer(Moschus berezovskii Flerov,1929)(FMD),which are considered as special skin glands,secrete a mixture of sebum,lipids,and proteins into the musk pod.Together,these components form musk,which plays an important role in attracting females during the breeding season.However,the relationship between the musk glands and skin of Chinese FMD remains undiscovered.Here,the musk gland and skin of Chinese FMD were examined using histological analysis and RNA sequencing(RNA-seq),and the expression of key regulatory genes was evaluated to determine whether the musk gland is derived from the skin.Methods A comparative analysis of musk gland anatomy between juvenile and adult Chinese FMD was conducted.Then,based on the anatomical structure of the musk gland,skin tissues from the abdomen and back as well as musk gland tissues were obtained from three juvenile FMD.These tissues were used for RNA-seq,hematoxylin-eosin(HE)staining,immunohistochemistry(IHC),western blot(WB),and quantitative real-time polymerase chain reaction(qRT-PCR)experiments.Results Anatomical analysis showed that only adult male FMD had a complete glandular organ and musk pod,while juvenile FMD did not have any well-developed musk pods.Transcriptomic data revealed that 88.24%of genes were co-expressed in the skin and musk gland tissues.Kyoto Encyclopedia of Genes and Genomes(KEGG)signaling pathway analysis found that the genes co-expressed in the abdomen skin,back skin,and musk gland were enriched in biological development,endocrine system,lipid metabolism,and other pathways.Gene Ontology(GO)enrichment analysis indicated that the genes expressed in these tissues were enriched in biological processes such as multicellular development and cell division.Moreover,the Metascape predictive analysis tool demonstrated that genes expressed in musk glands were skin tissue-specific.qRT-PCR and WB revealed that sex-determining region Y-box protein 9(Sox9),Caveolin-1(Cav-1),and androgen receptor(AR)were expressed in all three tissues,although the expression levels differed among the tissues.According to the IHC results,Sox9 and AR were expressed in the nuclei of sebaceous gland,hair follicle,and musk gland cells,whereas Cav-1 was expressed in the cell membrane.Conclusions The musk gland of Chinese FMD may be a derivative of skin tissue,and Sox9,Cav-1,and AR may play significant roles in musk gland development.
基金This work was supported by the National Key R&D program of China(2018YFD0502204)the National Natural Science Foundation of China(31872962).
文摘Analysis of the intestinal microbiota and physiological parameters in mammalian infancy can reveal health status.In this study,we used a combination of molecular and immunochemical approaches to assess fecal microbiota as well as Cortisol(Cor),Triiodothyronine(T3),and immunoglobulin A(IgA)levels of young forest musk deer(FMD),from birth to one month after weaning(7 days of age–110 days of age).During development as the diet of FMD changes from consuming milk to eating plants,the richness and diversity of intestinal microbiota of young FMD increased significantly.Cor levels remained unchanged throughout early development while significantly increased after weaning,T3 and IgA initially were derived from milk during lactation,significantly decreased after weaning.Correlation network analysis showed that the community of food-oriented microbes were highly structured and that many genera were correlated.Overall,this study provides scientific insights into effective management strategies for the protection of FMD population.
基金supported by the National Natural Science Foundation of China (30870362)the State Forestry Administration of Chinathe Fundamental Research Funds for the Central Universities of China
文摘The forest musk deer(Moschus berezovskii) is one of the most endangered species in China.Over the past decades,extensive hunting and poaching have pushed the forest musk deer to the edge of extinction,and conservation biologists are presently pursuing scientific management plans to rescue this species.The major histocompatibility complex(MHC),a cluster of genes responsible for antigen presentation,is a highly polymorphic genomic region in vertebrates that has become a popular functional marker system for studying adaptive variation.In this study,we developed locus-specific genotyping primers for exon 2 fragments of one DRA gene and one DRB locus of the forest musk deer using a suite of comprehensive methods that included universal primer amplification,genome walking,single-strand conformation polymorphism(SSCP),heteroduplex(HD) profiling,and sequence analysis.Each forest musk deer showed no more than two sequences per locus,confirming the specificity of our primers.Genotyping with these primers allowed us to identify two DRA alleles and six DRB alleles in a captive breeding population of the Sichuan Musk Deer Breeding Institution.For the DRA locus,we found a slightly higher observed heterozygosity(N O =0.154) than expected(N E =0.143).In contrast,the DRB locus showed a significant heterozygote deficiency(N O =0.508;N E =0.761;P<0.05),which was probably due to inbreeding in the captive population.An obvious excess of nonsynonymous substitutions over synonymous was observed at the antigen-binding positions of the DRA and DRB loci,showing the presence of positive selection in the forest musk deer DR genes.Finally,generation of phylogenetic trees for the DRA and DRB sequences of the forest musk deer and other ruminants revealed that the DRA and DRB loci identified in this study had homologous relationships with the known ruminant DRA and DRB genes.Based on this analysis,and to facilitate future studies,we named these novel loci Mobe-DRA and Mobe-DRB3.
