P-203: Examination of FMR1 Gene Transcription and Protein Expression in Patients with Diminished Ovarian Reserve Reffered to Royan institute

Background: Diminished ovarian reserve (DOR) is a primary infertility disorder characterized by a reduction in the number and/or quality of oocytes, usually accompanied by high follicle-stimulating hormone (FSH) levels and regular menses. DOR aetiology factors are different, such as genetic factors, ageing, autoimmune disorders, adrenal gland impairment factors and iatrogenic causes, e.g. chemotherapy. Premature ovarian failure (POF) is related to DOR and both are diagnosed by high levels of FSH. Furthermore, DOR has been described as a feature of POF. The association between FMR1 premutations and the POF disease has suggested that the FMR1 gene acts as a risk factor for the POF and recently for the DOR. In this study mRNA and protein expression of FMR1 gene in granulosa cells of women with DOR were analyzed. Materials and Methods: Local ethical approval was gained for this study and informed consent was given by patients. Granulosa cells were collected from 30 infertile women referred to Royan Institute. Samples distributed into two groups(DOR patients and non-DOR patients) based on the number of follicles, FSH levels and the number of CGG repeat. The DOR patients group included samples that had higher levels of FSH(>11), number of CGG repeats between 50-200 and follicles number lower than three .Each group contained 15 samples. Using reverse transcription and quantitative real-time PCR reaction(qRT-PCR) methods, the expression profile of FMR1 gene was evaluated. Also expression of the FMR1 encoded protein(FMRP) was analyzed by immunocytochemistry on ovarian granulosa cells biopsy. Results: Significantly higher expression levels of FMR1 gene in granulosa cells of DOR patients compared to non-DOR patients was shown. FMR1 protein expression was also shown in granulosa cells of DOR patients. Conclusion: Our data suggest that within the permutation range, a strong correlation is observed between translation of FMR1 mRNA and the number of CGG repeats in premutation alleles.