Background: Black bone sheep was first discovered in Yunnan province of China in 1970, with unique black pigmentation on the body and internal organs. Endothelin 3(EDN3) has been known as a key gene causing hyperpigme...Background: Black bone sheep was first discovered in Yunnan province of China in 1970, with unique black pigmentation on the body and internal organs. Endothelin 3(EDN3) has been known as a key gene causing hyperpigmentation in black bone chicken, the Silky fowl.Methods: In this study, EDN3 was employed as a candidate gene for regulating black color pigmentation. First,EDN3 was cloned from sheep to obtain the full-length cDNA by using the rapid amplification of cDNA ends(RACE).Genomic EDN3 was screened and a total of thirty predicted single nucleotide polymorphisms(SNPs) were genotyped for allele and genotype frequency analysis in a case-control study involving two black bone sheep populations. Genomic copy number analysis of EDN3 in sheep was conducted to measure the variation in copy number. EDN3 expression levels were observed among the groups in adult liver, lymph node, and kidney tissues, as well as embryo kidney samples. Also, among the tissues of black bone and non-black bone sheep.Results: The size of the full-length cDNA was 1,578 bp, which included 426 bp of 5′-untranslated region(5′-UTR),an open reading frame(ORF) of 639 bp encoding a protein of 212 amino acids, and a 3′-UTR of 513 bp. Genotype and allele frequencies of all the discovered SNPs were found insignificantly different in black bone and non-black bone sheep(P > 0.05). Genomic copy number analysis of EDN3 in sheep revealed no significant difference between the two sheep groups. No significant variations were found in the adult liver and kidney embryo samples. However,the expression in lymph node and kidney tissue was significantly higher in black bone sheep than that in non-black bone sheep(P < 0.05). Significant variations in the EDN3 expression levels were observed among the tissues of nonblack bone sheep.Conclusions: The findings of the present study indicate that unlike in Silky chickens, EDN3 is not responsible for hyperpigmentation but may play a key functional role in immune and excretory systems of black bone sheep.展开更多
Background: The widely observed RNA-DNA differences(RDDs) have been found to be due to nucleotide alteration by RNA editing. Canonical RNA editing(i.e., A-to-I and C-to-U editing) mediated by the adenosine deaminases ...Background: The widely observed RNA-DNA differences(RDDs) have been found to be due to nucleotide alteration by RNA editing. Canonical RNA editing(i.e., A-to-I and C-to-U editing) mediated by the adenosine deaminases acting on RNA(ADAR) family and apolipoprotein B mRNA editing catalytic polypeptide-like(APOBEC)family during the transcriptional process is considered common and essential for the development of an individual.To date, an increasing number of RNA editing sites have been reported in human, rodents, and some farm animals;however, genome-wide detection of RNA editing events in sheep has not been reported. The aim of this study was to identify RNA editing events in sheep by comparing the RNA-seq and DNA-seq data from three biological replicates of the kidney and spleen tissues.Results: A total of 607 and 994 common edited sites within the three biological replicates were identified in the ovine kidney and spleen, respectively. Many of the RDDs were specific to an individual. The RNA editing-related genes identified in the present study might be evolved for specific biological functions in sheep, such as structural constituent of the cytoskeleton and microtubule-based processes. Furthermore, the edited sites found in the ovine BLCAP and NEIL1 genes are in line with those in previous reports on the porcine and human homologs, suggesting the existence of evolutionarily conserved RNA editing sites and they may play an important role in the structure and function of genes.Conclusions: Our study is the first to investigate RNA editing events in sheep. We screened out 607 and 994 RNA editing sites in three biological replicates of the ovine kidney and spleen and annotated 164 and 247 genes in the kidney and spleen, respectively. The gene function and conservation analysis of these RNA editing-related genes suggest that RNA editing is associated with important gene function in sheep. The putative functionally important RNA editing sites reported in the present study will help future studies on the relationship between these edited sites and the genetic traits in sheep.展开更多
Musk is a scarce and precious medical resource secreted by male forest musk deer(FMD).Current research to promote musk secretion in FMD has used almost exclusively hormone injections,but this approach can be detriment...Musk is a scarce and precious medical resource secreted by male forest musk deer(FMD).Current research to promote musk secretion in FMD has used almost exclusively hormone injections,but this approach can be detrimental to the health of FMD.In order to conserve this endangered species as much as possible while increasing the production of musk,this study first used bioinformatics methods to predict the function of quercetin,a flavonoid that promotes testosterone(T)production and prevents late-onset male hypogonadism.On the basis of good prediction effect,different concentrations of quercetin were added to the diet of FMD.The results showed that quercetin could change the levels of T,luteinizing hormone releasing hormone,luteinizing hormone,and estradiol,and regulate the structure of intestinal microorganisms and musk microorganisms of FMD.Moreover,there is a correlation among musk components,hormones,intestinal microorganisms,and musk microorganisms,which indicates that the production of musk may be regulated by these three at the same time,and the addition of quercetin with 800 mg per kg diet could significantly increase the yield of muscone(P<0.05),the most effective ingredient in musk.In addition,quercetin decreased the high level of cortisol during musk secretion,which may relieve the stress on FMD in this process.This may help to protect the health of FMD.Combined with the results of software prediction,we finally proposed a possible mechanism for the complex process of musk secretion in FMD with a view to providing ideas for further studies.展开更多
基金supported by National Nature Science Foundation of China(U1136605)the Innovation Base Cultivation and Development Projectresearch on Precise genetic modification in sheep(Z171100002217072)Program for Changjiang Scholar and Innovation Research Team in University(IRT1191)
文摘Background: Black bone sheep was first discovered in Yunnan province of China in 1970, with unique black pigmentation on the body and internal organs. Endothelin 3(EDN3) has been known as a key gene causing hyperpigmentation in black bone chicken, the Silky fowl.Methods: In this study, EDN3 was employed as a candidate gene for regulating black color pigmentation. First,EDN3 was cloned from sheep to obtain the full-length cDNA by using the rapid amplification of cDNA ends(RACE).Genomic EDN3 was screened and a total of thirty predicted single nucleotide polymorphisms(SNPs) were genotyped for allele and genotype frequency analysis in a case-control study involving two black bone sheep populations. Genomic copy number analysis of EDN3 in sheep was conducted to measure the variation in copy number. EDN3 expression levels were observed among the groups in adult liver, lymph node, and kidney tissues, as well as embryo kidney samples. Also, among the tissues of black bone and non-black bone sheep.Results: The size of the full-length cDNA was 1,578 bp, which included 426 bp of 5′-untranslated region(5′-UTR),an open reading frame(ORF) of 639 bp encoding a protein of 212 amino acids, and a 3′-UTR of 513 bp. Genotype and allele frequencies of all the discovered SNPs were found insignificantly different in black bone and non-black bone sheep(P > 0.05). Genomic copy number analysis of EDN3 in sheep revealed no significant difference between the two sheep groups. No significant variations were found in the adult liver and kidney embryo samples. However,the expression in lymph node and kidney tissue was significantly higher in black bone sheep than that in non-black bone sheep(P < 0.05). Significant variations in the EDN3 expression levels were observed among the tissues of nonblack bone sheep.Conclusions: The findings of the present study indicate that unlike in Silky chickens, EDN3 is not responsible for hyperpigmentation but may play a key functional role in immune and excretory systems of black bone sheep.
基金funded by the National Nature Science Foundation of China(U1136605)Ningxia Agricultural Breeding Program(NXNYYZ20150103)Program for Changjiang Scholar and Innovation Research Team in University(IRT1191)
文摘Background: The widely observed RNA-DNA differences(RDDs) have been found to be due to nucleotide alteration by RNA editing. Canonical RNA editing(i.e., A-to-I and C-to-U editing) mediated by the adenosine deaminases acting on RNA(ADAR) family and apolipoprotein B mRNA editing catalytic polypeptide-like(APOBEC)family during the transcriptional process is considered common and essential for the development of an individual.To date, an increasing number of RNA editing sites have been reported in human, rodents, and some farm animals;however, genome-wide detection of RNA editing events in sheep has not been reported. The aim of this study was to identify RNA editing events in sheep by comparing the RNA-seq and DNA-seq data from three biological replicates of the kidney and spleen tissues.Results: A total of 607 and 994 common edited sites within the three biological replicates were identified in the ovine kidney and spleen, respectively. Many of the RDDs were specific to an individual. The RNA editing-related genes identified in the present study might be evolved for specific biological functions in sheep, such as structural constituent of the cytoskeleton and microtubule-based processes. Furthermore, the edited sites found in the ovine BLCAP and NEIL1 genes are in line with those in previous reports on the porcine and human homologs, suggesting the existence of evolutionarily conserved RNA editing sites and they may play an important role in the structure and function of genes.Conclusions: Our study is the first to investigate RNA editing events in sheep. We screened out 607 and 994 RNA editing sites in three biological replicates of the ovine kidney and spleen and annotated 164 and 247 genes in the kidney and spleen, respectively. The gene function and conservation analysis of these RNA editing-related genes suggest that RNA editing is associated with important gene function in sheep. The putative functionally important RNA editing sites reported in the present study will help future studies on the relationship between these edited sites and the genetic traits in sheep.
基金supported by the Forestry Science and Technology Innovation Project of Shaanxi Province(grant number SXLK2021-0219,SXLK2020-0207).
文摘Musk is a scarce and precious medical resource secreted by male forest musk deer(FMD).Current research to promote musk secretion in FMD has used almost exclusively hormone injections,but this approach can be detrimental to the health of FMD.In order to conserve this endangered species as much as possible while increasing the production of musk,this study first used bioinformatics methods to predict the function of quercetin,a flavonoid that promotes testosterone(T)production and prevents late-onset male hypogonadism.On the basis of good prediction effect,different concentrations of quercetin were added to the diet of FMD.The results showed that quercetin could change the levels of T,luteinizing hormone releasing hormone,luteinizing hormone,and estradiol,and regulate the structure of intestinal microorganisms and musk microorganisms of FMD.Moreover,there is a correlation among musk components,hormones,intestinal microorganisms,and musk microorganisms,which indicates that the production of musk may be regulated by these three at the same time,and the addition of quercetin with 800 mg per kg diet could significantly increase the yield of muscone(P<0.05),the most effective ingredient in musk.In addition,quercetin decreased the high level of cortisol during musk secretion,which may relieve the stress on FMD in this process.This may help to protect the health of FMD.Combined with the results of software prediction,we finally proposed a possible mechanism for the complex process of musk secretion in FMD with a view to providing ideas for further studies.
