Variants Of Meiosis Arrest Female1(MARF1)in Chinese Han Women With Oocyte Meiosis Arrest

Background Oocyte meiosis arrest is one of the reasons of women infertility. At present, according to the morphology of the nuclear, when observed germinal vesicles without polar body, the oocyte is in its Gv stage, however, when neither germinal vesicles nor polar body can be seen, the oocyte is in its MI stage. In2012, Youqiang Su et al used N-ethyl-N-nitrosourea (ENU) to induce mice Marfl mutations. As a result, oocytes are arrested in Gv stage and female mice are infertile. Human meiosis arrest female1(MARF1) locates in16pl3.11, containing27exons that encode1742-amino acid peptide, which is86%homology with mice Marfl amino acids. It has3RNA Recognition Motifs (RRMs). To explore the relationship between human oocyte meiosis arrest and MARFlgene mutations, RRMs encoding exon8,11,12, as well as exons16to27were screened in this project.Methods Sequence analysis is conducted of MARF1gene in91Chinese Han patients with oocyte meiosis arrest, who were recruited in Reproductive Hospital Affiliated to Shandong University during2008to2013. The inclusion criteria include1. Age minor than40years old.3. Normal karyotype.2. Two or more IVF/ICSI cycles of oocyte maturation failure. Besides,200controls that had given birth to healthy babies were also included in this project. Genomic DNA was extracted from peripheral blood samples. Subsequently, the8th,11th,12th,16th to27th exons of MARF1gene were amplified using polymerase chain reaction (PCR) with specific primers. The PCR product was sequenced on an automated sequencer and compared to the sequence in NCBI and finally analyze whether there are abnormal alteration insertions and deletions.Results We found one new synonymous mutation, two new single-nucleotide polymorphisms (SNPs) and five known SNPs in MARF1. The newfound synonymous mutation is in exon26, c.4852G>A. Two mew SNPs are in exon21(c.3979A>T) and exon25(c.4715C>G) separately. There are no significant differences between case group and control. Five known SNPs are rs28670396, rs7185421, rs28509427, rs58075088and rs202242803.Conclusion There is no strong evidence indicating that MARF1gene is related to human oocyte maturation failure, however, enlarged sample size repetition and functional researches of the new synonymous mutation are still needed.