Search

This research was stated on the group of infertile patient taking part in in vitro fertilization. The whole group of 60 random patients was divided into two subgroups: 10 persons smoking cigarettes and 50 nonsmokers. The number of follicles obtained after controlled ovarian hiperstimulation, the number of mature and immature oocytes and concentration of 17beta-estradiol in serum at the hCG day were measured for all women. There was also calculated the number of granulosa cells and the relative expression of the gene encoding FSHR. Comparing all testing groups, in smokers the worse ovarian answer for hiper-stimulation was observed. It was characterized by both the lower number of follicles and oocytes and also the lower concentration of estradiol in serum at hCG day. There was also observed lower number of granulosa cells in fluid from all folicles at diameter > 12 mm in smoking patients compared with nonsmokers. Analizing the relative expression of gene coding FSHR in granulosa cells significant differences between these two groups were demonstrated. The evaluation of the expression level showed its reduction in smokers (2). To compare it the average level of expression in nonsmokers was 24. Smoking cigarettes is correlated with the lower number of granulosa cells and the lower expression level of the gene coding for folitropin receptor.

Unlabelled: FSH-R expression in granulosa cells varies during the course of ovarian ontogenesis, as well as, during each menstrual cycle. Expression of follicle-stimulating hormone receptors (FSH-R) on Sertoli cells of the testis and ovarian granulosa cells depend on many paracrine and autocrine factors. The modulation of FSH-R synthesis is accomplished via a number of mechanisms including regulation of the promoter activity. Little is known about factors involved in control of FSH-R transcription in different species. The aim of the present study was to investigate differences in the regulation of human and rat FSH-R promoter activity by E2F transcriptional factors., Material and Methods: The 5'-flanking regions of human and rat FSH receptor gene subcloned in the pGL3 plasmid were transiently transfected into cultured CHO cells and rat granulosa cells. Rat granulosa cells were obtained by puncturing ovaries from DES primed immature Sprague-Dawley rats. Promoter activity was determined by measuring firefly luciferase luminescence of the cell lysate. Transfection efficiency was normalized by the renilla luciferase activity generated by co-transfected pRL-CMV vector. In order to determine the influence of E2F1, E2F4 and E2F5, on FSH-R promoter activity, cells were transfected either with promoter construct alone or with its mixture with selected expression vector., Results: Rat FSH-R promoter construct (-1033/+6 bp) and human FSH-R promoter construct (-1485/-1 bp) were both active in transfected cells. Overexpression of E2F1 protein decreased both, human and rat wild type FSH-R promoter activity. Overexpression of E2F4 did not affect neither rat nor human FSH-R gene transcription. Expression vector for E2F5 increased both, human and rat, FSH-R promoter activity. Folds of increase were markedly higher in case of rat FSH-R construct transfection, comparing to human FSH-R promoter., Conclusions: Results suggest, that the E2F1 and E2F5 factors might play an opposite role in the regulation of FSH-R promoter activity. More pronounced stimulatory effect of E2F5 on the rat FSH-R can be explained by the presence of E2F site in the promoter. Since there is no E2F sensitive element in the human FSH-R promoter sequence, E2F1 and E2F5 can also indirectly influence FSH-R promoter activity.