Rhesus monkey whole genome expression microarray of endometrium obtained on days 2, 4 and 6 after ovulation in mated, proven conception cycles and non-mated, normo-ovulatory cycles in rhesus monkeys
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PROVIDER: PRJNA428507 | ENA |
REPOSITORIES: ENA
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Project description:Several lines of evidence indicate that there exists differential display of biochemical characteristics in endometrium during receptivity in the proven conception cycle as compared to corresponding secretory phases endometrium of non-conception cycles of fertile rhesus monkeys. In the present study, we examined the differential display in the whole genome expression of endometrium during days 2, 4 and 6 post-ovuation in proven conception cycles and normo-ovulatory, non-conception cycles in proven fertile rhesus monkeys. Female monkeys showing at least two consecutive ovulatory menstrual cycles of normal length were allocated to either of the two groups: group 1 in which females were allowed to cohabit with proven fertile males during days 8 to 16 of their menstrual cycles, and group 2 in which females were not allowed to cohabit. Vaginal smears were checked daily and the days of ovulation were assessed from immunopositive profiles of estradiol-17β and progesterone in peripheral serum samples. Endometrial samples were collected on days 2, 4 and 6 after ovulation from both groups of animals following laparotomy and fundal hysterectomy under ketamine anaesthesia. In group 1, only endometrial samples obtained from mated females who yielded viable, preimplantation-stage matched embyos on uterine flushing were used. In group 2, endometrial samples were obtained from females who showed normal ovulatory cycles. Tissue samples from both groups were washed in ice-cold PBS and were used for RNA extraction to run rhesus monkey whole genome expression array. On average, ~60% and ~68% genes of total number of hybridized genes were expressed in endometrial samples of groups 1 and 2, respectively. Group-wise hierarchical cluster analysis (HCA) yielded highest segregation between peri-implantation stage (groups 1b and 1c) and other (groups 2a-c and group 1a) samples with cluster distance (cd) 0.9, when cd=1.0 denotes complete segregation. On the other hand, least segregation was seen between groups 1a and 2a with cd 0.2, when cd=0 denotes complete aggregation. The number of common genes were less with the progress in days in group 1 (that is in proven conception cycle) unlike that in group 2 (that is in non-mated, normo-ovulatory group). Also, the number of common genes between groups for a given day after ovulation displayed a clear declining trend with increasing number of uncommon genes. Differential dipslay of regulated common genes among all the subgroups based on statistical analyses of variances in data sets obtained from whole genome expression arrays identified a total of fifteen (15) genes that displayed differential (>2-fold, p< 0.05) expression: ADCY5, ADIPOR1, AVEN, CHRND, FOXD3, GJD4, MAPK8IP3, MKS1, NNMT, NUP50, PALT1, PIGV, TGFBR2, TOX2, VWA5B1.
2022-01-31 | GSE108760 | GEO
Project description:The molecular and cellular processes required for development, function, and regression of the primate corpus luteum (CL) are poorly defined. We hypothesized that there are dynamic changes in gene expression occurring during the CL lifespan, which represent proteins and pathways critical to its regulation. Therefore, a genomic approach was utilized to systematically identify differentially expressed genes in the rhesus macaque CL during the luteal phase of natural menstrual cycles. CL were collected between days 3-5 (early stage), 7-8 (mid), 10-12 (mid-late), 14-16 (late), or 18-19 (very-late) after the midcycle LH surge. From the early through very-late stages, 3234 transcripts were differentially expressed, with 879 occurring from the early through late stages that encompass the processes of luteinization, maintenance, and functional regression. To characterize gene changes most relevant to these processes, ontology analysis was performed using the list of 879 differentially expressed transcripts. Four main groups of related genes were identified with relevance to luteal physiology including: 1) immune function; 2) hormone and growth factor signaling; 3) steroidogenesis; and 4) prostaglandin biosynthesis, metabolism, and signaling. A subset of genes representing each of the four major categories was selected for validation of microarray results by quantitative real-time PCR. Results in mRNA levels were similar between the two methodologies for 17 of 18 genes. Additionally, protein levels for 3 genes were determined by Western blot analysis to parallel mRNA levels. This database will facilitate the identification of many novel or previously underappreciated pathways that regulate the structure and function of the primate CL. Keywords: time course The sole experimental factor is the stage of the luteal phase when the CL was collected. There were five stages: 1) between days 3-5 post LH-surge (early stage), 2) 7-8 (mid stage), 3) 10-12 (mid-late stage), 4) 14-16 (late stage), and 5) 18-19 (very-late stage). The early CL are undergoing luteinization, the mid are fully functional CL that are at their peak progesterone producing capacity, the mid-late stage is a transitionary period where CL are still producing significant quantities of progesterone but are nearing the time when luteolysis initiates in non-conception cycles, the late stage corresponds to CL undergoing functional regression (cessation of progesterone secretion), and the very-late stage (menses) is when the structural remodeling and apoptosis associated with luteolysis is occurring. Measuring mRNA levels in CL collected during these periods provides a comprehensive analysis of the primate transcriptome throughout CL lifespan. There were 20 biological replicates: n = 4 per stage, 5 stages in total. The early CL were used as the baseline reference for gene expression.
2008-02-08 | E-GEOD-10367 | biostudies-arrayexpress
Project description:Luteolysis of the corpus luteum (CL) during non-fertile cycles involves a cessation of progesterone (P4) synthesis (functional regression) and subsequent structural remodeling. The molecular processes responsible for initiation of luteal regression in the primate CL are poorly defined. Therefore, a genomic approach was utilized to systematically identify differentially expressed genes in the rhesus macaque CL during spontaneous luteolysis. CL were collected prior to (days 10-11 post-LH surge, mid-late [ML] stage) or during (days 14-16, late stage) functional regression. Based on P4 levels, late stage CL were subdivided into functional late (FL, serum P4 > 1.5 ng/ml) and functionally-regressed late (FRL, serum P4 < 0.5 ng/ml) groups (n=4 CL/group). Total RNA was isolated, labeled and hybridized to Affymetrix genome microarrays that contain elements representing the entire rhesus macaque transcriptome. With the ML stage serving as the baseline, there were 681 differentially expressed transcripts (>2-fold change; p< 0.05) that could be categorized into three primary patterns of expression: 1) increasing from ML through FRL, 2) decreasing from ML through FRL, and 3) increasing ML to FL, followed by a decrease in FRL. Ontology analysis revealed potential mechanisms and pathways associated with functional and/or structural regression of the macaque CL. Quantitative real-time PCR was used to validate microarray expression patterns of 13 genes with the results being consistent between the two methodologies. Protein levels were found to parallel mRNA profiles in 4 of 5 differentially expressed genes analyzed by Western blot. Thus, this database will facilitate the identification of mechanisms involved in primate luteal regression. Keywords: time course plus functional state of corpus luteum The first experimental factor is the stage of the luteal phase when the CL was collected. There were two stages: 1) between days 10-12 post LH-surge (mid-late stage), and 2) days 14-16 (late stage). The mid-late stage is a transitionary period where CL are still functional (i.e., producing significant quantities of P4) but are nearing the time when luteolysis initiates in non-conception cycles, and the late stage corresponds to the normal period when functional regression (cessation of P4 secretion) occurs in non-conception cycles. The second experimental factor is the functional state of the CL which was determined by measuring serum concentrations of P4 at the time of CL collection. Late stage CL with serum P4 levels > 1.5 ng/ml were considered to be functional, and those with serum P4 < 0.5 ng/ml were considered to have completed functional regression. Measuring mRNA levels in CL collected from these groups provides a comprehensive analysis of the primate transcriptome throughout the normal period of functional regression in the macaque CL. There were 12 biological replicates: n = 4 per group, 3 groups in total. The mid-late CL were used as the baseline reference for gene expression.
2008-10-27 | E-GEOD-12807 | biostudies-arrayexpress
Project description:Endocervical mucus changes play a key role in regulating fertility throughout the menstrual cycle and in response to hormonal contraceptives. Non-human primates (NHP) provide the most translational animal model for reproductive tract studies, as they have hormonally-regulated menstrual cycles and mucus changes, similar to women. We used TMT labelling and LC-LC/MS to compare the proteins found in the mucus of the rhesus macaque to the mucus of the human endocervix. We found 3,048 total proteins present in both rhesus mucus and human mucus, and of these, 57% showed a similar expression pattern. An even higher similarity occurred in the top 500 most prevalent proteins, with overlap in 341 (68%) proteins. Mucin MUC5B was the most highly expressed mucin protein (top 10 expressed proteins in both) but other key proteins related to mucus structure were present in both samples. We find that the mucus proteome of the endocervical mucus is highly conserved in NHP and women. This supports use of the NHP model system for studies of the endocervix and trials of novel fertility treatments targeting the cervix.
2021-05-21 | PXD021710 | Pride
Project description:The molecular and cellular processes required for development, function, and regression of the primate corpus luteum (CL) are poorly defined. We hypothesized that there are dynamic changes in gene expression occurring during the CL lifespan, which represent proteins and pathways critical to its regulation. Therefore, a genomic approach was utilized to systematically identify differentially expressed genes in the rhesus macaque CL during the luteal phase of natural menstrual cycles. CL were collected between days 3-5 (early stage), 7-8 (mid), 10-12 (mid-late), 14-16 (late), or 18-19 (very-late) after the midcycle LH surge. From the early through very-late stages, 3234 transcripts were differentially expressed, with 879 occurring from the early through late stages that encompass the processes of luteinization, maintenance, and functional regression. To characterize gene changes most relevant to these processes, ontology analysis was performed using the list of 879 differentially expressed transcripts. Four main groups of related genes were identified with relevance to luteal physiology including: 1) immune function; 2) hormone and growth factor signaling; 3) steroidogenesis; and 4) prostaglandin biosynthesis, metabolism, and signaling. A subset of genes representing each of the four major categories was selected for validation of microarray results by quantitative real-time PCR. Results in mRNA levels were similar between the two methodologies for 17 of 18 genes. Additionally, protein levels for 3 genes were determined by Western blot analysis to parallel mRNA levels. This database will facilitate the identification of many novel or previously underappreciated pathways that regulate the structure and function of the primate CL. Keywords: time course
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