Expression data from early preimplantation Bovine embryos
Ontology highlight
ABSTRACT: The oviduct is a specialized organ playing crucial roles in the success of early reproductive events and it provides an optimal microenvironment for early embryonic development. However, changes in oviductal environment due to estrus synchronization and superovulation hormonal treatments and subsequent influence on embryos transcriptome profile are not yet investigated. For that, the objective of this study was to investigate differences in developmental rate and transcriptome profile of bovine blastocysts cultured under superovulation or synchronization oviductal environment. Influence of oviductal environment on transcriptome abundance of produced blastocysts was examined using the Affymetrix GeneChip Bovine Genome Array. Eighteen Simmental heifers were synchronized, superovulated and artificially inseminated, then nine of them were flushed at day 2 by transvaginal endoscopic means and 2-cell stage embryos were recovered and endoscopicaly transferred to only synchronized recipients. The remaining half of superovulated heifers and the synchronized recipients were flushed at day 7 to collect blastocysts which developed either in superovulated or synchronized oviduct
Project description:The oviduct is a specialized organ playing crucial roles in the success of early reproductive events and it provides an optimal microenvironment for early embryonic development. However, changes in oviductal environment due to estrus synchronization and superovulation hormonal treatments and subsequent influence on embryos transcriptome profile are not yet investigated. For that, the objective of this study was to investigate differences in developmental rate and transcriptome profile of bovine blastocysts cultured under superovulation or synchronization oviductal environment.
Project description:The objective of this study was to examine the effect of the presence of a single or multiple embryo(s) on the transcriptome of the bovine oviduct. In Experiment 1, cyclic (non-bred, n = 6) and pregnant (artificially inseminated, n = 11) heifers were slaughtered on Day 3 after estrus, and the ampulla and isthmic regions of the oviduct ipsilateral to the corpus luteum were separately flushed. Oviductal epithelial cells from the isthmus region, in which all oocytes/embryos were located, were snap-frozen for microarray analysis. In Experiment 2, heifers were divided into cyclic (non-bred, n = 6) or pregnant (multiple embryo transfer, n = 10) groups. In vitro-produced presumptive zygotes were transferred endoscopically to the ipsilateral oviduct on Day 1.5 post estrus (n = 50 zygotes per heifer). Heifers were slaughtered on Day 3 and oviductal isthmus epithelial cells were recovered for RNA sequencing. Microarray analysis in Experiment 1 failed to detect any difference in the transcriptome of the oviductal isthmus induced by the presence of a single embryo. In Experiment 2, following multiple embryo transfer, RNA sequencing revealed 278 differentially expressed genes of which 123 were up- and 155 were down-regulated in pregnant heifers. Most of the down-regulated genes were related to immune function. Five samples from pregnant heifers and five control, cyclic heifers were analysed
Project description:The objective of this study was to examine the effect of the presence of a single or multiple embryo(s) on the transcriptome of the bovine oviduct. In Experiment 1, cyclic (non-bred, n = 6) and pregnant (artificially inseminated, n = 11) heifers were slaughtered on Day 3 after estrus, and the ampulla and isthmic regions of the oviduct ipsilateral to the corpus luteum were separately flushed. Oviductal epithelial cells from the isthmus region, in which all oocytes/embryos were located, were snap-frozen for microarray analysis. In Experiment 2, heifers were divided into cyclic (non-bred, n = 6) or pregnant (multiple embryo transfer, n = 10) groups. In vitro-produced presumptive zygotes were transferred endoscopically to the ipsilateral oviduct on Day 1.5 post estrus (n = 50 zygotes per heifer). Heifers were slaughtered on Day 3 and oviductal isthmus epithelial cells were recovered for RNA sequencing. Microarray analysis in Experiment 1 failed to detect any difference in the transcriptome of the oviductal isthmus induced by the presence of a single embryo. In Experiment 2, following multiple embryo transfer, RNA sequencing revealed 278 differentially expressed genes of which 123 were up- and 155 were down-regulated in pregnant heifers. Most of the down-regulated genes were related to immune function. Transcriptional profiles of oviductal isthmus epithelial cells from cyclic and pregnant heifers were generated by sequencing of total RNA on the Illumina HiSeq 2500 platform
Project description:The objective of this study was to examine the effect of the presence of a single or multiple embryo(s) on the transcriptome of the bovine oviduct. In Experiment 1, cyclic (non-bred, n = 6) and pregnant (artificially inseminated, n = 11) heifers were slaughtered on Day 3 after estrus, and the ampulla and isthmic regions of the oviduct ipsilateral to the corpus luteum were separately flushed. Oviductal epithelial cells from the isthmus region, in which all oocytes/embryos were located, were snap-frozen for microarray analysis. In Experiment 2, heifers were divided into cyclic (non-bred, n = 6) or pregnant (multiple embryo transfer, n = 10) groups. In vitro-produced presumptive zygotes were transferred endoscopically to the ipsilateral oviduct on Day 1.5 post estrus (n = 50 zygotes per heifer). Heifers were slaughtered on Day 3 and oviductal isthmus epithelial cells were recovered for RNA sequencing. Microarray analysis in Experiment 1 failed to detect any difference in the transcriptome of the oviductal isthmus induced by the presence of a single embryo. In Experiment 2, following multiple embryo transfer, RNA sequencing revealed 278 differentially expressed genes of which 123 were up- and 155 were down-regulated in pregnant heifers. Most of the down-regulated genes were related to immune function.
Project description:The objective of this study was to examine the effect of the presence of a single or multiple embryo(s) on the transcriptome of the bovine oviduct. In Experiment 1, cyclic (non-bred, n = 6) and pregnant (artificially inseminated, n = 11) heifers were slaughtered on Day 3 after estrus, and the ampulla and isthmic regions of the oviduct ipsilateral to the corpus luteum were separately flushed. Oviductal epithelial cells from the isthmus region, in which all oocytes/embryos were located, were snap-frozen for microarray analysis. In Experiment 2, heifers were divided into cyclic (non-bred, n = 6) or pregnant (multiple embryo transfer, n = 10) groups. In vitro-produced presumptive zygotes were transferred endoscopically to the ipsilateral oviduct on Day 1.5 post estrus (n = 50 zygotes per heifer). Heifers were slaughtered on Day 3 and oviductal isthmus epithelial cells were recovered for RNA sequencing. Microarray analysis in Experiment 1 failed to detect any difference in the transcriptome of the oviductal isthmus induced by the presence of a single embryo. In Experiment 2, following multiple embryo transfer, RNA sequencing revealed 278 differentially expressed genes of which 123 were up- and 155 were down-regulated in pregnant heifers. Most of the down-regulated genes were related to immune function.
Project description:Greater progesterone (P4) concentrations during the follicular growth have been associated with greater quality of embryos produced by superovulated cows and heifers. However, it is yet to be determined the mechanisms by which oocyte exposure to greater P4 concentrations improves early embryo quality. The objective of this study was to evaluate the impact of different P4 concentrations during superovulation on the transcriptome profile of early bovine embryos. A total of 63 post-puberty Holstein heifers were randomly assigned into two experimental groups: High P4 (n = 31) and Low P4 (n = 32). Heifers received a pre-synchronization protocol followed by a protocol of superovulation that included the allocated P4 treatment. Embryo collection was performed 7 days post artificial insemination and embryos were evaluated for stage of embryonic development and grades of quality. Embryos graded as good/excellent quality (High P4: n = 27; Low P4: n = 27) were randomly allocated in 3 biological replicates per treatment group, balanced for stage of embryonic development. Transcriptome analysis suggested that exposure to different concentrations of P4 during superovulation promote changes in gene expression of 7 days old embryos that may be related to their developmental competence. These modifications are associated with downregulation of beta-estradiol pathway, upregulation of trophoblast-related genes and downregulation of WNT signalling pathway. Our results suggest a potential sensitivity of future embryos to follicular P4 levels but do not allow to conclude if the effect is from the oocyte, the oviduct, or the uterus response to P4.
Project description:The hypothesis tested was that the uterine environment of lactating cows would affect conceptus gene expression. Approximately 65-75 days post-partum (dpp) the estrous cycles of non-lactating (dried off immediately post partum: n=12) and lactating (n=13) cows were synchronized and on Day 7 a high quality blastocyst derived from superovulated heifers was transferred. A control group of maiden heifers (n=8) were synchronized, inseminated to a standing heat and slaughtered on the same day as non-lactating and lactating recipients (Day 19; estrus=Day 0). The ipsilateral uterine horn was flushed with 10 ml PBS and the conceptus, when present, and uterine luminal fluid (ULF) snap frozen in liquid nitrogen prior to analysis. Gene expression analysis of the conceptus was performed by RNA sequencing analysis while amino acid (aa) composition of ULF was determined by High Performance Liquid Chromatography (HPLC). Eight differentially expressed genes (DEGs) were identified between conceptuses recovered from non-lactating cows versus heifers while 269 DEGs (100 up-regulated and 169 down-regulated) were identified between conceptuses recovered from lactating cows compared to heifers. The aa, alanine, glycine, serine and threonine, arginine, leucine and valine, were significantly lower in abundance in ULF recovered from heifers compared to both non-lactating or lactating cows. Glutamic acid, glutamine and lysine concentrations were lowest in heifers compared to both cow groups. This study demonstrates that exposure of a grade one embryo to a uterine environment that has been exposed to the metabolic stresses associated with lactation modifies the transcriptome of the conceptus and aa composition of the ULF.
Project description:The aim of this study was to compare the transcriptome of the different regions of the oviduct between pregnant and cyclic heifers. After synchronizing crossbred beef heifers, those in standing oestrus (=Day 0) were randomly assigned to cyclic (non bred, n=6), or pregnant (artificially inseminated, n=11) groups. They were slaughtered on Day 3 and both oviducts from each animal were isolated and cut in half to separate ampulla and isthmus. Each portion was flushed to confirm the presence of an oocyte/embryo and was then opened longitudinally and scraped to obtain epithelial cells which were snap-frozen. Oocytes and embryos were located in the isthmus of the oviduct ipsilateral to the corpus luteum. Microarray analysis of oviductal cells revealed that proximity to the corpus luteum did not affect the transcriptome of the isthmus, irrespective of pregnancy status. However, 2287 genes were differentially expressed (P<0.01) between the ampulla and isthmus of the oviduct ipsilateral to the corpus luteum. Gene ontology revealed that the main biological processes overrepresented in the isthmus were synthesis of nitrogen, lipids, nucleotides, steroids and cholesterol as well as vesicle-mediated transport, cell cycle, apoptosis, endocytosis and exocytosis, whereas cell motion, motility and migration, DNA repair, calcium ion homeostasis, carbohydrate biosynthesis and regulation of cilium movement and beat frequency were overrepresented in the ampulla. In conclusion, large differences in gene expression were observed between the isthmus and ampulla that reflect morphological and functional characteristics of each segment. Fifteen samples including 5 pregnant contralateral isthmus heifers, 5 cyclic contralateral isthmus heifers, and 5 pregnant ipsilateral ampulla heifers. Also, ten samples from GSE74593 were reanalyzed with these samples. The full set of processed data are linked at the bottom of the Series record.
Project description:The aim of this study was to compare the transcriptome of the different regions of the oviduct between pregnant and cyclic heifers. After synchronizing crossbred beef heifers, those in standing oestrus (=Day 0) were randomly assigned to cyclic (non bred, n=6), or pregnant (artificially inseminated, n=11) groups. They were slaughtered on Day 3 and both oviducts from each animal were isolated and cut in half to separate ampulla and isthmus. Each portion was flushed to confirm the presence of an oocyte/embryo and was then opened longitudinally and scraped to obtain epithelial cells which were snap-frozen. Oocytes and embryos were located in the isthmus of the oviduct ipsilateral to the corpus luteum. Microarray analysis of oviductal cells revealed that proximity to the corpus luteum did not affect the transcriptome of the isthmus, irrespective of pregnancy status. However, 2287 genes were differentially expressed (P<0.01) between the ampulla and isthmus of the oviduct ipsilateral to the corpus luteum. Gene ontology revealed that the main biological processes overrepresented in the isthmus were synthesis of nitrogen, lipids, nucleotides, steroids and cholesterol as well as vesicle-mediated transport, cell cycle, apoptosis, endocytosis and exocytosis, whereas cell motion, motility and migration, DNA repair, calcium ion homeostasis, carbohydrate biosynthesis and regulation of cilium movement and beat frequency were overrepresented in the ampulla. In conclusion, large differences in gene expression were observed between the isthmus and ampulla that reflect morphological and functional characteristics of each segment.
Project description:Use of combined estrus synchronization/superovulation treatments (SS) alters ovarian and en-dometrial gene expression patterns, impairing follicle and oocyte growth, fertilization, and em-bryo development. Since the impact of SS-treatments on the transcriptome of the surviving em-bryos remains undisclosed, we hereby examined gene expression changes in day 6 blastocysts that survived a brief regimen of synchronization treatment combined with superovulation. Embryos were surgically collection at day 6 after AI and transcriptome analysis performed on blas-tocyst-stage embryos with good morphology to disclose differentially expressed genes (DEGs) between groups at P-value < 0.05 and </> 1.5-fold change. Compared with the blastocysts from control (untreated) sows, the blastocysts from SS treated sows had moderate gene expression changes, with 7 pathways disrupted with a total of 10 transcripts affected, including metabolic RDH10 and SPTLC2 gene upregulation, and downregulation of oxidation related GSTK1 and GSTO1 genes. These gene expression alterations may suggest suboptimal embryo quality and could depress the embryos' response to oxidative stress, thereby impairing subsequent embryo development. These and previous findings call for avoidance of SS-treatments in embryo transfer programs.