Expression of mRNAs for DNA-methyltransferases and Histone Deacetylases in Granulosa Cells and Follicular Fluid of Women Undergoing in vitro Fertilization

Background: Gene products involved in reproduction frequently undergo post-transcriptional modifications by DNA methylation and histone acetylation. The aim was to assess the predictive value of gene expression levels of DNA methyltransferases (DNMTs) and histone deacetylases (HDACs) in patients treated with in vitro fertilization (IVF). In a prospective, l clinical study 31 consecutive patients with male (n=17) or female (n=14) infertility diagnoses were enrolled. Granulosa cells (GCs) and follicular fluid (FF) were obtained at the oocytes retrieval during IVF. mRNA levels of DNMT1, DNMT3a, DNMT3b and HDAC5, HDAC6 were measured in GCs and FF by quantitative RT-PCR using ROCHE Lightcycler 480. The outcome measures were the number of oocytes retrieved, mature oocytes and viable embryos, as well as chemical and clinical pregnancy. Results: : It was demonstrated that genes for DNMTs and HDACs could be detected in nearly equal amount in GC and FF, however, only the DNMT3a transcript in FF correlated with that in GC (r=0.478, p<0.033). Moreover, FF DNMT3a was significantly higher in the pregnant (N=9) than in the non-pregnant (N=22) patients (p<0.016), and HDAC6 in GC was significantly related to the number of oocytes retrieved (r=0.413, p<0.026), MII oocytes (r= 0.383, p<0.040) and viable embryos (r=0.413, p<0.025). Conclusions: : In our clinical setting the expression of mRNA for FF DNMT3a and for GC HDAC6 has the potential to assess IVF outcome.