Exploring The Potential Association Between CapG Gene And Hypoxia Adaptation In Tibetan Pigs

Tibetan pigs,which live on the plateau all year round,have strong resilience and hypoxic adaptation,which are good research materials for conducting research on plateau adaptation.To investigate the association between hypoxia adaptation mechanism and CapG gene in Tibetan pigs,this study was conducted on Tibetan pigs and York pigs living in Linzhi,Tibet(average altitude 2900 m)for a long time,using the phenotypic data of blood physiological indicators of the two pig breeds as the starting point,combined with the expression of CapG at DNA level,m RNA level and protein level,using real-time fluorescence quantitative PCR(Quantitative Real-time PCR(RT-q PCR),generation sequencing technology,immunohistochemistry(Immuno Histo Chemistry,IHC),immunoblot analysis(Western blot,WB)and blood physiological indexes measurement were used to explore the function of porcine CapG gene through the in-depth To further understand the genetic basis of hypoxic adaptation in highland animals,with the aim of providing a basis for the laws of highland physiological responses and molecular mechanisms of genetic adaptation in animals.1.Blood physiological indicators were measured in Tibetan and York pigs.The results showed that the values of five indexes,namely red blood cell count(RBC),hemoglobin count(HGB),red blood cell specific volume(HCT),mean red blood cell hemoglobin content(MCH)and mean red blood cell hemoglobin concentration(MCHC),were highly significant higher in Tibetan pigs than in York pigs(P < 0.01);The coefficient of variation of red blood cell distribution width(RDW)in York pigs was extremely significantly higher than that in Tibetan pigs(P < 0.01);Other physiological indices were not significantly different between the two pig breeds(P > 0.05).2.Single Nucleotide Polymorphism(SNP)was detected using generation sequencing technology to explore the key functional loci of CapG gene in Tibetan and York pigs.By analyzing the sequencing results of 5’ flanking region and CDS region,it was found that the CapG gene was mutated only in Tibetan pigs,with six mutation sites in 5’ flanking region,A-556 C,C-489 T,T-459 C,A-429 G,C-274 T and A-212 G,and no sense mutation was found in CDS region.After transcription factor prediction,transcription factors were found to be altered at the C-489 T,C-274 T,and A-212 G loci and were associated with hypoxic adaptation and blood oxygen transport.3.In this experiment,the expression of CapG gene in heart,liver,spleen,lung and kidney tissues of Tibetan pigs and York pigs was examined by RT-q PCR.The m RNA expression of CapG gene in the heart,liver,spleen,lung and kidney was significantly higher in the Tibetan pigs than in the Yorke pigs(P< 0.01).4.Immunohistochemistry was used to localize the protein expression of CapG protein in the lung tissues of Tibetan pigs and Yorkshire pigs.The results showed that CapG protein was widely expressed in the lung tissues of Tibetan pigs and Yorke pigs,mainly in the cytoplasm and cell membrane;the expression level of CapG protein in the lungs of Yorke pigs was very significantly higher than that of Tibetan pigs(P < 0.01).5.The expression of CapG protein in the heart,liver,spleen and lung tissues of Tibetan pigs and York pigs was examined by Western blot technique.The results showed that the expression of CapG protein in the heart,liver and spleen of Tibetan pigs was not significantly(P > 0.05)different from that of Yorke pigs,and the overall trend was higher in Yorke pigs than Tibetan pigs;in the lung tissues,the expression of CapG protein was significantly higher in Yorke pigs than Tibetan pigs(P < 0.05).In conclusion,CapG gene is an important candidate gene for regulating hypoxic adaptation in Tibetan pigs,and plays an important role in regulating hypoxic adaptation in Tibetan pigs on the plateau.This study provides a theoretical basis for resource conservation and utilization of Tibetan pigs,breeding of plateau breeds and disease control,and has certain reference value for the development of plateau livestock economy.