Effect Of Carbonic Anhydrase 4 Gene On PH In Chicken Fibroblast Culture Medium

Carbonic anhydrase 4(CA4)is a member of the carbonic anhydrase family and catalyzes the reversible reaction of carbon dioxide and water to produce bicarbonate and hydrogen ions,mainly affecting the pH of the cell environment.Currently,research based on CA4 is mainly focused on the lungs,skeletal muscles,and cancers.There is less research on reproductive development.Studies have shown that CA4 expression in chicken seminal epithelial cells,and high expression in the uterine vaginal junction,suggesting that CA4 regulation of the female poultry gland acid-base environment.In this study,chicken embryo fibroblasts(DF-1)was used as a model to study the pH of CA4 to DF-1 cell culture medium and L-lactic acid by cell culture,immunofluorescence,immunohistochemistry and qRT-PCR using RNAi method.effect.The main findings are as follows:(1)Screening for the SNP site in the exon segment of chicken CA4 gene and found six SNP sites,among which there are four SNPs whose codon changes caused amino acid changes.The C/T mutation occurs at base position 620,mutation of this site causes the mutation of the proline to leucine,and the T721 mutation occurs at the base position 721.This site mutates the tryptophan to arginine.A/T mutation occurs at base position 796.This site mutation causes the encoded lysine to be mutated to a stop codon and the 847 th base undergoes a G/T mutation.This site mutation results in the mutation of the encoded glycine to a cysteine.Amino acid.(2)Both CA4 and MCT4 genes are expressed in DF-1 cells and localized to the cell membrane.Therefore,DF-1 cells can be used to study the function of CA4 gene.(3)After the CA4 gene expression was decreased,the pH of the cell culture solution decreased,while the content of L-lactic acid in the DF-1 cell culture solution was significantly decreased(p<0.05).