Comparison of gene expression between male and female blastocysts
Ontology highlight
ABSTRACT: Male embryos are reported to develop faster than female in the preimplantation stage. Therefore, male and female embryos can be considered phenotypically different as early as the preimplantation stage. Employing our sexing system of enhanced green fluorescent protein (EGFP) tagging X chromosomes, we compared the global gene expression pattern of male and female embryos at the blastocyst stage using DNA microarray. Keywords= sex differentiation Keywords= preimplantation embryo Keywords= genomic imprinting Keywords: comparative hybridization
Project description:It is basically understood that male and female development is initiated by gonad differentiation into either testis or ovary. However, male embryos are reported to develop faster than female during preimplantation, implying sex differences at this stage. To learn more about when sex differentiation begins, we compared the global gene expression pattern of male and female embryos at the blastocyst stage. First, Blastocyst samples were sexed, using a novel method for non-invasive sexing of preimplantation stage mouse embryos by tagging the X chromosome with an EGFP transgene, Next, gene expression patterns of the male and female were compared using DNA microarray.
Project description:Male embryos are reported to develop faster than female in the preimplantation stage. Therefore, male and female embryos can be considered phenotypically different as early as the preimplantation stage. Employing our sexing system of enhanced green fluorescent protein (EGFP) tagging X chromosomes, we compared the global gene expression pattern of male and female embryos at the blastocyst stage using DNA microarray. Experiment Overall Design: samples were collected from three independent preparations and the experiments were triplicated. Scanned microarray results were processed with Feature Extraction software (ver. 7.5, Agilent). ãThe hybridization experiments were duplicated in a reciprocal labeling manner to reduce dye integration bias, and total of six hybridizations were carried out for the entire analysis.ã Combining plural array results and statistical analyses were carried out by Luminator software (Rosetta).
Project description:It is basically understood that male and female development is initiated by gonad differentiation into either testis or ovary. However, male embryos are reported to develop faster than female during preimplantation, implying sex differences at this stage. To learn more about when sex differentiation begins, we compared the global gene expression pattern of male and female embryos at the blastocyst stage. First, Blastocyst samples were sexed, using a novel method for non-invasive sexing of preimplantation stage mouse embryos by tagging the X chromosome with an EGFP transgene, Next, gene expression patterns of the male and female were compared using DNA microarray. ï¼³amples were collected from three independent preparations and the experiments were triplicated. Scanned microarray results were processed with Feature Extraction software (ver. 7.5, Agilent). ãThe hybridization experiments were duplicated in a reciprocal labeling manner to reduce dye integration bias, and total of six hybridizations were carried out using each 22K-1 and 22K-2 array.ã Combining plural array results and statistical analyses were carried out by Luminator software (Rosetta).
Project description:Sexual dimorphism in mammals is mostly attributable to sex-related hormonal differences in fetal and adult tissues; however, this may not be the sole determinant. Though genetically-identical for autosomal chromosomes, male and female preimplantation embryos could display sex-specific transcriptional regulation which can only be attributted to the differences in sexual chromosome dosage. We used microarrays to analyze sex-related transcriptional differences at the blastocyst stage.
Project description:Sexual dimorphism in mammals is mostly attributable to sex-related hormonal differences in fetal and adult tissues; however, this may not be the sole determinant. Though genetically-identical for autosomal chromosomes, male and female preimplantation embryos could display sex-specific transcriptional regulation which can only be attributted to the differences in sexual chromosome dosage. We used microarrays to analyze sex-related transcriptional differences at the blastocyst stage. Day 7 bovine in vitro produced bovine blastocysts produced with sorted semen from 3 different bulls. Pooled RNA from 60 blastocysts of one sex and produced with one bull was used per chip. Three replicates of each sex per bull. In total, 18 Bovine GeneChip (Affymetrix) were used (3 replicates X 3 bulls X 2 sexes).
Project description:Environmental estrogens may affect epigenetic programming as early as the period of preimplantation development. Therefore, we analyzed the effects of continuous gestational estradiol-17β (E2) exposure on male and female embryos. A low dose, close to the no-observed effect level (NOEL - 10 µg E2/kg body weight(bw)/d), a high dose (1000 µg E2/kg bw/d) and carrier only, as control group, was fed to sows from insemination until sampling at day 10 of pregnancy, respectively. 36 samples (n = 5-7 per treatment group and sex) were analyzed by high throughput sequencing.In the high dose group, RNA-sequencing of single embryos revealed 982 differentially expressed genes (DEG) in the female but none in the male blastocysts. Moreover, 62 and 3 DEG were found in female and male embryos of the NOEL dose group, respectively. Thus, maternal E2 treatment during early pregnancy affected gene expression of the embryos at day 10, potentially constituting the basis for long-term adverse effects.
Project description:Biological bases for sexual differences in the brain exist in a wide range of vertebrate species, including the chicken. We examined whether sexually dimorphic gene expression in the brain precedes gonadal differentiation. Using the Affymetrix GeneChip® Chicken Genome Array, we identified many female- and male-enhanced genes that are differentially expressed in sex-specific brains from stage 29 chicken embryos. We postulate that these genes have potential roles in the sexual differentiation of neural function and development in chickens. Experiment Overall Design: All analyses were performed in triplicate. The Affymetrix data from male and female brain samples at stage 29 were generated. The male and female brain samples by genetic sexing were pooled and homogenized. Pooled samples ranged from 5 to 8 embryos. 4g of total RNA from the pooled samples was used for labeling. Probe synthesis from total RNA samples, hybridization, detection, and scanning were performed according to standard protocols from Affymetrix.
Project description:Endocannabinoid signaling plays a key role in multiple events in female reproduction. In this investigation, we discovered an interesting phenomenon that mice with either elevated or silenced endocannabinoid signaling show similar defects in multiple pregnancy events, including preimplantation embryo development. To unravel the underlying mechanisms, microarray studies were was conducted using RNAs collected from WT, Cnr1-/- and Faah-/- mouse blastocysts on day 4 of pregnancy. The results show that about 100 genes showed unidirectional changes under either elevated or silenced endocannabinoid signaling. Analysis of functional grouping of these genes revealed that multiple biological functions and pathways are affected under aberrant endocannabinoid signaling, including cell migration. Several genes from the microarray data were confirmed by quantitative RT-PCR. Cell motility assays validated the predicted compromised cell migration in Cnr1-/- and Faah-/- trophoblast stem cells. This study provides molecular basis for biphasic effects of endocannabinoid signaling in female reproduction To generate Cnr1-/-, Faah-/- and WT embryos, mutant and WT females were mated with males of the same genotypes. On day 4 of pregnancy at 1500h, embryos were collected from oviducts and uteri of pregnant females, because Cnr1-/- and Faah-/- females show aberrant oviductal transport of embryos. As expected, WT embryos were mostly at the blastocyst stage and all of them were collected from uteri. However, Cnr1-/- and Faah-/- females had higher percentages of embryos at the morula stage, and some of them were still in the oviduct. Since gene expression profiles greatly vary depending on the embryonic development stage, morulae were not included in gene expression analysis; only RNAs from blastocysts were used to better understand the effects solely arising from disrupted cannabinoid signaling in blastocysts.
Project description:Pigs are important animals for agricultural and biomedical research, and its assisted reproductive technologies are urgently needed to be improved. Determining the underlying mechanism of epigenetic reprogramming in the early stage of preimplantation embryos derived from in vitro fertilization (IVF), parthenogenesis (PA) and androgenesis (AG), will not only contribute to assisted reproduction technologies of pigs but also will shed light into early human development. We generated 3D chromatin profiles for pig somatic cells, IVF, PA and AG preimplantation embryos. We found that the chromosomes in the pig preimplantation embryos are enriched for superdomains, i.e., the larger than 10M compartment domains, which are much rarer in mice. Wheras, p(s) curves, compartments and TADs, are largely conserved in somatic cells, and are gradually establishing during preimplantation embryogenesis. In the uniparental pig embryos, the establishment of chromatin architecture was highly asynchronized at all levels from IVF embryos, and a remarkably strong decompartmentalization was observed during zygotic genome activation (ZGA). Finally, chromosomes originating from oocytes always establish TADs faster than chromosomes originating from sperm, both before and during ZGA.
Project description:Pigs are important animals for agricultural and biomedical research, and its assisted reproductive technologies are urgently needed to be improved. Determining the underlying mechanism of epigenetic reprogramming in the early stage of preimplantation embryos derived from in vitro fertilization (IVF), parthenogenesis (PA) and androgenesis (AG), will not only contribute to assisted reproduction technologies of pigs but also will shed light into early human development. We generated 3D chromatin profiles for pig somatic cells, IVF, PA and AG preimplantation embryos. We found that the chromosomes in the pig preimplantation embryos are enriched for superdomains, i.e., the larger than 10M compartment domains, which are much rarer in mice. Wheras, p(s) curves, compartments and TADs, are largely conserved in somatic cells, and are gradually establishing during preimplantation embryogenesis. In the uniparental pig embryos, the establishment of chromatin architecture was highly asynchronized at all levels from IVF embryos, and a remarkably strong decompartmentalization was observed during zygotic genome activation (ZGA). Finally, chromosomes originating from oocytes always establish TADs faster than chromosomes originating from sperm, both before and during ZGA.