Differential gene expression in human granulosa cells from recombinant FSH versus human menopausal gonadotropin ovarian stimulation protocols

Abstract

BackgroundThe study was designed to test the hypothesis that granulosa cell (GC) gene expression response differs between recombinant FSH and human menopausal gonadotropin (hMG) stimulation regimens.MethodsFemales < 35 years-old undergoing IVF for tubal or male factor infertility were prospectively randomized to one of two stimulation protocols, GnRH agonist long protocol plus individualized dosages of (1) recombinant (r)FSH (Gonal-F) or (2) purified human menopausal gonadotropin (hMG; Menopur). Oocytes were retrieved 35 h post-hCG, and GC were collected. Total RNA was extracted from each GC sample, biotinylated cRNA was synthesized, and each sample was run on Human Genome Bioarrays (Applied Microarrays). Unnamed genes and genes with <2-fold difference in expression were excluded.ResultsAfter exclusions, 1736 genes exhibited differential expression between groups. Over 400 were categorized as signal transduction genes, ~180 as transcriptional regulators, and ~175 as enzymes/metabolic genes. Expression of selected genes was confirmed by RT-PCR. Differentially expressed genes included A kinase anchor protein 11 (AKAP11), bone morphogenetic protein receptor II (BMPR2), epidermal growth factor (EGF), insulin-like growth factor binding protein (IGFBP)-4, IGFBP-5, and hypoxia-inducible factor (HIF)-1 alpha.ConclusionsResults suggest that major differences exist in the mechanism by which pure FSH alone versus FSH/LH regulate gene expression in preovulatory GC that could impact oocyte maturity and developmental competence.

Effects of Urinary and Recombinant Gonadotropins on In Vitro Maturation Outcomes of Mouse Preantral Follicles

“…[17][18][19][20][21] No differences were observed in the relative expression of 3 genes among the 4 groups, in Original magnification was 60Â and scale bars represent 100 mm. In the hMG group, granulosa cells were denser than other groups on day 10 (C).…”

“…The 3 genes, octamer-binding transcription factor 4 (Oct4), bone marrow protein 15 (Bmp15), and growth differentiation factor 9 (Gdf9) were selected in this study, as oogenesis-specific genes, which had been studied for high expression during oocyte maturation. [17][18][19][20][21][22] Quantitative RT-PCR of these 3 genes was carried out using SYBR green PCR mix (Qiagen, Valencia, California). The amplification program included 45 cycles of denaturation at 95 C for 15 seconds, annealing at 58 C for 20 seconds, and extension at 72 C for 30 seconds.…”

Gene expression profiling of granulosa cells from PCOS patients following varying doses of human chorionic gonadotropin

“…These observations were further supported by subsequent studies that measured global gene expression of cumulus cells, which was concluded to be a non-invasive test for oocyte/embryo quality [2,3,15,41]. Granulosa and cumulus cell gene expression profiling has also been used to compare the effects of recombinant FSH vs. human menopausal gonadotrophin (hMG) [6,14], floating vs. cumulus granulosa cells [22], cumulus cells from lean vs. overweight-obese PCOS patients [21], and ovarian reserve status in young women with diminished ovarian reserve [39].…”

Factors Influencing the Biochemical Markers for Predicting Mammalian Oocyte Quality

“…In support of this finding, Brannian et al [67] used the DNA microarray technology to discover 1,736 differentially expressed genes in a similar experimental setup. Of these genes, more than 400 were categorized as signal transduction genes, ~180 were categorized as transcriptional regulators, and ~175 were categorized as enzymes/ metabolic genes.…”