[Objective]The aim of this study was to verify the joint regulatory relationship between DNA methylation and mRNA expression of candidate genes in the brain tissue of piglets infected by Glaesserella parasuis(GPS),in order to provide a theoretical basis for revealing the apparent pathogenic mechanism of meningitis caused by GPS in piglets.[Method]Six 28-day-old weaned piglets were randomly divided into control and GPS groups.The piglets in GPS group were intraperitoneally injected with 1 mL of 2 × 109 CFU/mL GPS SH0165 solution,and the piglets in control group were intraperitoneally injected with the same amount of normal saline,and the brain of the piglets were collected 7 days later to extract DNA and RNA.The mRNA expression of 4 candidate genes(LYPD1,PITPNM1,SYP and ACVR1B)were detected by Real-time quantitative PCR,and bisulfite sequencing PCR(BSP)and methylation-specific PCR(MSP)were used to detect the DNA methylation changes of 4 genes in brain tissues of piglets before and after GPS infection.[Result]This study successfully applied the MSP method to DNA methylation sequencing of genes,making it not limited to fixed-point detection of methylation sites.The results of Real-time quantitative PCR showed that compared with control group,the mRNA expression of LYPD1,PITPNM1,SYP and ACVR1B genes in GPS group were significantly or extremely significantly down-regulated(P＜0.05 or P＜O.01).The results of DNA methylation sequencing showed that the DNA methylation level of 4 genes were up-regulated except for ACVR1B gene.[Conclusion]Except for ACVR1B gene,the DNA methylation of LYPD1,PITPNM1 and SYP genes was inversely associated with mRNA expression,and the DNA methylation changes in brain of piglet after GPS infection had different regulatory patterns for the expression of 4 candidate genes.
维普
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