Project description:The endometrium provides optimal conditions for the transport of sperm to the oviduct, to the site of fertilization, and later on for the reception of the embryo. To study these changes on the level of gene expression, a messenger RNA expression profiling of endometrium tissue samples collected from 19 cyclic heifers at five stages of the estrous cycle (days 0, 3.5, 12, 18, 20) was performed. RNA was extracted from these tissue samples and analyzed with a custom-made bovine oviduct and endometrium (BOE) cDNA array. The cDNAs present on the array were derived from several previously conducted differential gene expression studies of bovine endometrium between different stages of the estrous cycle, during early pregnancy, and from studies of bovine oviduct epithelial cells. In all of these studies cDNAs of differentially expressed genes were identified using a combination of subtracted cDNA libraries and cDNA array hybridization. 1,440 cDNA fragments are located on the array. Twenty radioactively labeled cDNA samples (n=4 for each cycle stage) were hybridized with BOE arrays. Raw data were normalized using the BioConductor package vsn. Keywords: time course of gene expression in bovine endometrium during estrous cycle
Project description:The endometrium provides optimal conditions for the transport of sperm to the oviduct, to the site of fertilization, and later on for the reception of the embryo. To study these changes on the level of gene expression, a messenger RNA expression profiling of endometrium tissue samples collected from 19 cyclic heifers at five stages of the estrous cycle (days 0, 3.5, 12, 18, 20) was performed. RNA was extracted from these tissue samples and analyzed with a custom-made bovine oviduct and endometrium (BOE) cDNA array. The cDNAs present on the array were derived from several previously conducted differential gene expression studies of bovine endometrium between different stages of the estrous cycle, during early pregnancy, and from studies of bovine oviduct epithelial cells. In all of these studies cDNAs of differentially expressed genes were identified using a combination of subtracted cDNA libraries and cDNA array hybridization. 1,440 cDNA fragments are located on the array. Twenty radioactively labeled cDNA samples (n=4 for each cycle stage) were hybridized with BOE arrays. Raw data were normalized using the BioConductor package vsn. Keywords: time course of gene expression in bovine endometrium during estrous cycle Nineteen German Fleckvieh (Simmental) heifers were slaughtered at four different days of the estrous cycle, four animals at day 0, four at day 3.5, three at day 12 and eight animals at day 18. Four of the day 18 animals showed high and the other four low serum progesterone (P4) levels, respectively. This resulted in five groups of endometrial tissue samples corresponding to five stages of the estrous cycle.
Project description:The bovine endometrium secretes proteins across the estrous cycle that vary (in both protein identity and abundance) to be in synchrony with luminal reproductive events. To uncover the identity of these proteins, transcriptomics was performed to understand the secretory capability of the bovine endometrium through the distinct phases of the estrous cycle in cows. This dataset should serve as a baseline for understanding the profile of luminal secretory outputs across the estrous cycle, setting the stage for functional interpretation of how the uterus supports and directs biological events ranging from sperm transport to preimplantation embryo development.
Project description:The aim of the present study is to determine possible roles of chemokines in regulating bovine endometrial function during early pregnancy. A 15 K bovine oligo DNA microarray detected 344 and 1336 differentially expressed genes (>2-fold; P<0.05) in the bovine endometrium of 15 and 18 days of pregnancy compared with these days of the estrous cycle (cyclic, n=4). The expression of six chemokines, including CCL2, CCL8, CCL11, CCL14, CCL16 and CXCL10 was higher in the endometrium of early pregnancy than that in the cyclic stage.
Project description:The endometrium plays a crucial role in the reproductive organs in the aspect of embryo-maternal communication and pregnancy. This study investigated transcriptome profiles of endometrial cells stimulated with PBS, LPA and LPA in combination with IFNt. LPA, one of the signaling molecule, is locally produced and released from the bovine endometrium during estrous cycle and early pregnancy. The highest concentration of LPA and expression of its active receptor (LPAR1) were detected in bovine endometrium at the time of maternal recognition of pregnancy, when the conceptus announces its presence by increased IFNt production. Using transcriptomic approach we compared the influence of LPA and LPA together with IFNt on the gene expression profiles in bovine endometrial cells.
Project description:Although somatic cell nuclear transfer (SCNT) cloning is more efficient in bovine than in all other species tested so far, there is a high rate of pregnancy failure that has been linked to structural and functional abnormalities of the placenta. We tested the hypothesis that these changes may originate from disturbed embryo-maternal interactions in the pre-implantation period. Therefore, we evaluated the transcriptome response of the endometrium to SCNT embryos (produced from five different donor cell cultures) as compared to embryos derived from in vitro fertilization (IVF). SCNT embryos and IVF embryos were cultured under identical conditions to the blastocyst stage (Day 8) and transferred to recipients. The recipients were slaughtered at day 18 of pregnancy and the uterus was recovered. Pregnancy was verified by the presence of at least one normally developed embryo. Transcriptome profiling of endometrium samples using a custom cDNA microarray covering transcripts expressed in the endometrium and/or oviduct epithelium revealed 58 transcripts that were differently abundant between endometrium samples from SCNT vs. IVF pregnancies. Prominent examples are NR2F2 (encoding the orphan nuclear receptor COUP-TFII) and GJA1 (encoding connexin 43). Both transcripts are known to play important roles in placentation and were significantly less abundant in endometrium from SCNT vs. IVF pregnancies. These findings suggest that placental failure in bovine clone pregnancies may originate from abnormal embryo-maternal communication already in the pre- or peri-implantation period. Endometrium transcriptome profiles may serve as a novel readout to evaluate SCNT embryos for their ability to induce pregnancy with a functional placenta. Keywords: response to different embryos Nineteen German Fleckvieh (Simmental) heifers were slaughtered at day 18 of pregnancy. Cycle-synchronized recipient heifers received either IVP or SCNT embryos at day 7 of the estrous cycle. Animals were slaughtered at day 18. Endometrial (intercaruncular) tissue samples were obtained from 10 pregnant animals after transfer of IVP embryos and from 9 pregnant animals after transfer of SCNT embryos.
Project description:We investigated differential gene expression profiles of endometrium during the mid-luteal phase of the estrous cycle between repeat breeding (RB) and normally fertilized cows using microarray analysis. Caruncular (CAR) and intercaruncular (ICAR) endometrium of both ipsilateral and contralateral uterine horns to the corpus luteum were collected from RB (inseminated at least three times but not pregnant) and normally fertile cows on Day 15 of the estrous cycle. Global gene expression profiles of these endometrial samples were analyzed with a 15K custom-made oligo-microarray in cattle. Microarray analysis revealed that 405 and 397 genes were differentially expressed in CAR and ICAR of the ipsilateral uterine horn of RB, respectively when compared with normal cows. In contralateral uterine horn, 443 and 257 differentially expressed genes were identified in the CAR and ICAR of RB, respectively when compared with normal cows. In the analysis of whole uterine (combining the above four portions), RB cows showed up-regulation of 37 genes including PRSS2, GSTA3, PIPOX, CNGA1 and IGG1C and down-regulation of 39 genes including CHGA, KRT35, THBS4, CPXM2 and PRF1 compared with normal cows. Our results may suggest that local regulation of molecular mechanisms in each uterine portion contributes to normal uterine physiology.
Project description:At implantation the endometrium undergoes dramatic modifications necessary for its physical interactions with the trophoblast as well as the development of the conceptus. We aim to identify endometrial factors and pathways essential for a successful implantation in the caruncular [C] and the intercaruncular [IC] areas in cattle. Using a bovine oligo-array, expression profiles were established at day 20 of the estrous cycle or pregnancy (implantation) showing 446 and 1295 differentially expressed genes (DEG) in [C] and [IC] areas respectively. The impact of the conceptus was higher on the immune response function in [C] but more prominent on the regulation of metabolism function in [IC]. Keywords: Fluorescence Microarray
Project description:The endometrium plays a crucial role in the reproductive organs in the aspect of embryo-maternal communication and pregnancy. This study investigated transcriptome profiles of endometrial cells stimulated with PBS, LPA and LPA in combination with IFNt. LPA, one of the signaling molecule, is locally produced and released from the bovine endometrium during estrous cycle and early pregnancy. The highest concentration of LPA and expression of its active receptor (LPAR1) were detected in bovine endometrium at the time of maternal recognition of pregnancy, when the conceptus announces its presence by increased IFNt production. Using transcriptomic approach we compared the influence of LPA and LPA together with IFNt on the gene expression profiles in bovine endometrial cells. A total of nine normally cycling Holstein/Polish Black and White (75/25% respectively) cows were used in this study. Global transcriptional profiling was performed using co-cultured stromal and epithelial cells (ratio - 3:1) isolated from bovine endometrium. Three experimental conditions (control (PBS), LPA and LPA plus IFNt) with three replicates per condition were prepared. Total RNAs were extracted from 9 pooled samples (n=3 for each sample) amplified and hybridized onto Affymetrix microarrays.
Project description:At implantation the endometrium undergoes dramatic modifications necessary for its physical interactions with the trophoblast as well as the development of the conceptus. We aim to identify endometrial factors and pathways essential for a successful implantation in the caruncular [C] and the intercaruncular [IC] areas in cattle. Using a bovine oligo-array, expression profiles were established at day 20 of the estrous cycle or pregnancy (implantation) showing 446 and 1295 differentially expressed genes (DEG) in [C] and [IC] areas respectively. The impact of the conceptus was higher on the immune response function in [C] but more prominent on the regulation of metabolism function in [IC]. Keywords: Fluorescence Microarray 18 intercaruncular samples, 18 caruncular Samples. Comparisons of cycle (d20) and pregnancy (d20).