Project description:Current knowledge of the molecular regulation of the blastocyst implantation event has been largely derived from studies in the mouse that requires ovarian estrogen for initiation of the implantation event. However, there are species such as the hamster, guinea pig, pig, horse, rhesus monkey and perhaps the human where the blastocyst implantation event initiates only in the progesterone-primed uterus. Despite this fundamental difference in the requirement of ovarian hormones in initiating blastocyst implantation among species, efforts to identify gene networks relevant for the blastocyst implantation event in progesteroneM-bM-^@M-^Sdependent species are limited. In this study, cDNA prepared from RNAs of day 5 blastocyst implantation and interimplantation sites were hybridized with mouse and human oligonucleotide microarray platforms to discern the transcriptional networks underlying the regulation of blastocyst implantation in hamsters. Compared with the inter-implantation site, blastocyst implantation sites showed upregulation and downregulation of a sizable number of genes by both cross-species arrays.The merit of the cross-species hybridization and reliability of the identified up- and down-regulated genes at the implantation sites were validated by detecting differential expression of a few randomly selected genes from both arrays by real-time PCR. Function gene ontology and pathway analysis revealed that differentially expressed genes are associated with several biological events and molecular pathways that are likely to be taking place at the blastocyst implantation site. This is the first study that identified the differential gene expression profile at the blastocyst implantation site of the hamsters, and revealed molecular pathways that are possibly associated with the progesterone-dependent blastocyst implantation process. We used microarrays to detail the expression differences in hamster implantation and interimplantation sites. Total RNAs were prepared from the hamster day 5 blastocyst implantation and interimplantation sites. Three sets of RNAs were isolated from three different animals and were subjected to microarray analysis using Affymetrix mouse and human array platforms.
Project description:Current knowledge of the molecular regulation of the blastocyst implantation event has been largely derived from studies in the mouse that requires ovarian estrogen for initiation of the implantation event. However, there are species such as the hamster, guinea pig, pig, horse, rhesus monkey and perhaps the human where the blastocyst implantation event initiates only in the progesterone-primed uterus. Despite this fundamental difference in the requirement of ovarian hormones in initiating blastocyst implantation among species, efforts to identify gene networks relevant for the blastocyst implantation event in progesteroneM-bM-^@M-^Sdependent species are limited. In this study, cDNA prepared from RNAs of day 5 blastocyst implantation and interimplantation sites were hybridized with mouse and human oligonucleotide microarray platforms to discern the transcriptional networks underlying the regulation of blastocyst implantation in hamsters. Compared with the inter-implantation site, blastocyst implantation sites showed upregulation and downregulation of a sizable number of genes by both cross-species arrays.The merit of the cross-species hybridization and reliability of the identified up- and down-regulated genes at the implantation sites were validated by detecting differential expression of a few randomly selected genes from both arrays by real-time PCR. Function gene ontology and pathway analysis revealed that differentially expressed genes are associated with several biological events and molecular pathways that are likely to be taking place at the blastocyst implantation site. This is the first study that identified the differential gene expression profile at the blastocyst implantation site of the hamsters, and revealed molecular pathways that are possibly associated with the progesterone-dependent blastocyst implantation process. We used microarrays to detail the expression differences in hamster implantation and interimplantation sites. Total RNAs were prepared from the hamster day 5 blastocyst implantation and interimplantation sites. Three sets of RNAs were isolated from three different animals and were subjected to microarray analysis using Affymetrix mouse and human array platforms.
Project description:Current knowledge of the molecular regulation of the blastocyst implantation event has been largely derived from studies in the mouse that requires ovarian estrogen for initiation of the implantation event. However, there are species such as the hamster, guinea pig, pig, horse, rhesus monkey and perhaps the human where the blastocyst implantation event initiates only in the progesterone-primed uterus. Despite this fundamental difference in the requirement of ovarian hormones in initiating blastocyst implantation among species, efforts to identify gene networks relevant for the blastocyst implantation event in progesterone–dependent species are limited. In this study, cDNA prepared from RNAs of day 5 blastocyst implantation and interimplantation sites were hybridized with mouse and human oligonucleotide microarray platforms to discern the transcriptional networks underlying the regulation of blastocyst implantation in hamsters. Compared with the inter-implantation site, blastocyst implantation sites showed upregulation and downregulation of a sizable number of genes by both cross-species arrays.The merit of the cross-species hybridization and reliability of the identified up- and down-regulated genes at the implantation sites were validated by detecting differential expression of a few randomly selected genes from both arrays by real-time PCR. Function gene ontology and pathway analysis revealed that differentially expressed genes are associated with several biological events and molecular pathways that are likely to be taking place at the blastocyst implantation site. This is the first study that identified the differential gene expression profile at the blastocyst implantation site of the hamsters, and revealed molecular pathways that are possibly associated with the progesterone-dependent blastocyst implantation process. We used microarrays to detail the expression differences in hamster implantation and interimplantation sites.
Project description:Current knowledge of the molecular regulation of the blastocyst implantation event has been largely derived from studies in the mouse that requires ovarian estrogen for initiation of the implantation event. However, there are species such as the hamster, guinea pig, pig, horse, rhesus monkey and perhaps the human where the blastocyst implantation event initiates only in the progesterone-primed uterus. Despite this fundamental difference in the requirement of ovarian hormones in initiating blastocyst implantation among species, efforts to identify gene networks relevant for the blastocyst implantation event in progesterone–dependent species are limited. In this study, cDNA prepared from RNAs of day 5 blastocyst implantation and interimplantation sites were hybridized with mouse and human oligonucleotide microarray platforms to discern the transcriptional networks underlying the regulation of blastocyst implantation in hamsters. Compared with the inter-implantation site, blastocyst implantation sites showed upregulation and downregulation of a sizable number of genes by both cross-species arrays.The merit of the cross-species hybridization and reliability of the identified up- and down-regulated genes at the implantation sites were validated by detecting differential expression of a few randomly selected genes from both arrays by real-time PCR. Function gene ontology and pathway analysis revealed that differentially expressed genes are associated with several biological events and molecular pathways that are likely to be taking place at the blastocyst implantation site. This is the first study that identified the differential gene expression profile at the blastocyst implantation site of the hamsters, and revealed molecular pathways that are possibly associated with the progesterone-dependent blastocyst implantation process. We used microarrays to detail the expression differences in hamster implantation and interimplantation sites.
Project description:Embryo implantation is a complex process which involves biochemical and physiological interactions between an implantation-competent blastocyst and a receptive uterus. However, the exact biochemical changes of uterine fluid, uterus, and plasma during peri-implantation remain unclear. This study aims to characterize the biochemical and metabolic changes that occur during the peri-implantation period of early pregnancy, using mice as an animal model. Gas chromatography-mass spectrometry was used to analyze the metabolite profiles of the uterus, uterine fluid, and maternal plasma at pre-implantation and implantation. The multivariate analyses, ANOVA and Tukey's HSD test, were applied to detect significant changes in metabolites and metabolic pathways. The metabolic networks were reconstructed in silico based on the identified metabolites and KEGG metabolic framework. Between pre-implantation day 1 and day 4, dramatic metabolic changes were observed in the uterine fluid that could be important for blastocyst development and protection against the harsh uterine environment. Palmitoleic acid, fumaric acid, and glutaric acid changed levels at day 4 in the uterus, suggesting that they may be associated with endometrial receptivity. Both the uterus and maternal plasma showed profound changes in cellular metabolism at the early implantation period, including upregulation of branched-chain amino acids and intermediates of one-carbon metabolism, an upregulation of glyoxylate and dicarboxylate metabolism, and downregulation of aerobic respiration; all of which could be involved in the regulation of the maternal-fetal interface, alternative nutrient utilization, and energy preservation for implantation as well as later placentation and fetal development to ensure successful embryo implantation.