Study Of Imprinting And Expression Of Arfip2Gene In Mouse

We screen the candidate imprinted genes in the mouse genome by two kindsof prediction model and numerous candidate genes were found, one of which isArfip2. Previous research about Arfip2has focused on the function of the protein,and the expression pattern of Arfip2has not been reported. So here we analyzedthe expression pattern of the gene in middle and later periods of mice embryonicdevelopment, and simply studied the imprinting status and regulationmechanism of the gene. Above these laid a foundation for further study Arfip2function.Bio-information suggested that the mouse Arfip2gene spans6.2Kb onchromosome7. Its coding protein is ADP ribosylation factor interacting protein2, belong to Arfaptin family. Arfip2is a conserved evolution gene. The structureof its protein functional domain is very similar to the BAR structure domain. Inthis paper, the imprinted status of Arfip2was analyzed by SNP direct sequencing.The results indicated that the gene was not an imprinting gene in E15.5mice’smain organizations (brain, tongue, heart, lungs, liver, kidneys and placenta).After that, we studied in detail the expression pattern of the gene in middle andlater periods of mice embryonic development by in situ hybridization andreal-time quantitative PCR technique. Results showed that the gene isspecifically expressed in the brain from E9.5to E11.5; its expression is mainlydistributed in the telencephalon, midbrain and rhomb encephalon. Arfip2wasubiquitously expressed in the embryo at E12.5day. Abundant signals weredetected in brain, tongue and liver and the signals are relatively weak in theheart, lungs and kidneys at E12.5. The expression of Arfip2widely existed in thebrain, tongue, lungs, liver, kidney and thymus, and in the heart is still very lowat E15.5. The results of real-time quantitative PCR exhibited that there is highlevel expression of Arfip2gene in the brain, tongue, lung, liver in later periodsof mice embryonic development. From E12.5to E18.5, the gene expression inthe heart gradually reduce, expression increased in the kidney. In other tissues,the expressions of the gene present a downward trend. Finally, we examined themethylation level of CpG Island in Arfip2gene promoter region. Bisulfitesequencing analysis revealed that Arfip2promoter region is methylation-free atE15.5, which means that the expression of the gene is not subject to regulationof DNA methylation at E15.5.