Project description:Microarray analysis of the transcriptome in the primate corpus luteum during chorionic gonadotropin administration simulating early pregnancy.
Project description:C. Stötzel, J. Plöntzke, W. Heuwieser & S. Röblitz. Advances in modeling of the bovine estrous cycle: synchronization with PGF2α. Theriogenology 78, 7 (2012).
Our model of the bovine estrous cycle is a set of ordinary differential equations which generates hormone profiles of successive estrous cycles with several follicular waves per cycle. It describes the growth and decay of the follicles and the corpus luteum, as well as the change of the key reproductive hormones, enzymes and processes over time. In this work we describe recent developments of this model towards the administration of prostaglandin F2α. We validate our model by showing that the simulations agree with observations from synchronization studies and with measured progesterone data after single dose administrations of synthetic prostaglandin F2α.
Project description:To explore chorionic gonadotropin (CG)-regulated gene expression in the primate corpus luteum (CL), adult female rhesus macaques were treated with a model of simulated early pregnancy (SEP). Total RNA was isolated from individual CL and hybridized to AffymetrixM-bM-^DM-" GeneChip Rhesus Macaque Genome Arrays The level of 1192 transcripts changed expression > 2-fold (one-way ANOVA, FDR correction; P<0.05) during SEP when compared to Day 10 untreated controls, and the majority of changes occurred between Days 10 and 12 of SEP. To compare transcript levels between SEP rescued and regressing CL, previously banked rhesus GeneChip array data from the mid- to late and very late luteal phase were analyzed with time-matched intervals in SEP. Comparing RMA-normalized transcripts from the natural cycle with those from luteal rescue revealed 7677 transcripts changing in expression pattern >2 fold (one-way ANOVA, FDR correction; P<0.05) between the two groups. Clustering of samples revealed that the SEP samples possessed the most related transcript expression profiles. Regressed CL (days 18-19, around menses) were the most unlike all other CL. The most affected KEGG pathway was Steroid Biosynthesis, and most significantly absent pathways following SEP treatment includes groups of genes whose products promote cell-death. By further comparing the genome-wide changes in luteal gene expression during rescue in SEP, with those in CL during luteolysis in the natural menstrual cycle, it is possible to identify key regulatory pathways promoting fertility. Simulated early pregnancy (SEP) treatment was begun on day 9 as Duffy and Stouffer (1997) by treatment of females with recombinant human chorionic gonadotropin (hCG; NovarelM-bM-^DM-", Ferring Pharmaceuticals Inc. Parsippany, NJ, USA) in increasing dosages (15, 30,45,90,180,360,720,1440, and 2880 IU) twice daily by intramuscular injection. CL were collected by laparotomy on days 10, 12, 15, and 18, representing 1, 3, 6 and 9 days of hCG treatment (n=4 CL/day). Additionally, luteal day 10 untreated CL were collected to serve as baseline controls for SEP CL. All CL were dissected away from luteal tissue, sectioned, and snap-frozen in liquid nitrogen and stored at -80M-BM-0C until RNA and protein isolation by TRIzolM-BM-. extraction (Invitrogen, Carlsbad, CA, USA) according to manufacturerM-bM-^@M-^Ys protocols.
Project description:Observational data suggest that women conceiving without a corpus luteum are at higher risk of developing preeclampsia. While the underlying mechanisms remain unclear, the absence of corpus luteum-derived secretory products may be a contributing factor. This study investigates whether the plasma proteome differs between women who conceive with or without a corpus luteum and examines the relationship with mode of conception. Plasma samples from 12 participants were collected at three time points: first trimester, third trimester, and postpartum. The cohort included women who conceived unassisted (UC) after infertility, via artificial cycle frozen embryo transfer (AC FET), or natural cycle frozen embryo transfer (NC FET). 36 plasma protein samples were analyzed using mass spectrometry-based proteomics to compare the proteome of women who conceived with and without a corpus luteum, across different conception methods and pregnancy stages. In total, 528 proteins were quantified. No differentially expressed plasma proteins were identified between women with and without a corpus luteum. However, 15 proteins showed differential expression between UC and FET at all time points, with Bonferroni-corrected p<9.47 × 10E-5 and FC≥|2|. Several altered proteins, including PAPPA and ANG, were linked to preeclampsia. SERPINA7 was differentially detected when comparing time points within the unassisted conception method. No significant differences were detected between AC FET and NC FET. This pilot study revealed a unique proteomic signature associated with mode of conception. The findings suggest biologically plausible candidate proteins for further testing. Validation in larger cohorts or with alternative proteome analysis technologies is needed
Project description:Early temporal transcriptome responses of the bovine mid-cycle corpus luteum to prostaglandin F2 alpha indicates cytokine activation