Unknown,Transcriptomics,Genomics,Proteomics

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Silencing or amplification of endocannabinoid signaling in blastocysts via CB1 compromises trophoblast cell migration

ABSTRACT: 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.

ORGANISM(S): Mus musculus

SUBMITTER: Minoru Ko

PROVIDER: E-GEOD-36399 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Project description:In present, interspecies cloning and interspecies-pregnancy were studied for endangered species rescue. However, the low implantation and survival ratio, spontaneous abortion, and unknown reason embryos absorption are the common and difficult problems of interspecies-pregnancy. In order to discover the mechanism of interspecies-pregnant failure and find ways to overcome the xeon-pregnant obstacles, we chosen the rat embryos pregnant in mouse uterus as a interspecies-pregnancy model. Three groups were set, mouse embryos to mouse recipients (MM) as control group, rat embryos to mouse recipients (RM), and rat and mouse embryos to mouse recipients together (RMM) as experiment groups. The former studies showed that rat embryos live no longer than day 7 of mouse pregnancy (D7). Our results showed that rat embryos survived to D7, and still existed to day 9 of mouse pregnancy (D9) in RM group. Surprisingly, the rat embryos survived to day 13 of the mouse gestation (D13) in RMM group. Microarray analysis was used to detect the global-gene expression profile changes of the whole implantation sites among the three groups at D7 and D9. By this way, we screened out the genes promoting the implanted rat embryos development in a mouse uterus which helped the rat embryos survive to D13 in RMM group compared with RM group, and the genes hindering the rat embryos development in a mouse uterus which prevented rat embryos living longer than D7 in RM group and D13 in RMM group compared with MM group. These findings provide insights into the mechanism of interspecies pregnant failure and new idea for interspecies pregnant studies. Experiment Overall Design: microarray was use to screen the genes among the day 7 and day 9 implantation sites of rat embryos implantation sites in a mouse uterus between rat embryos transfer to mouse recipients and rat embryos transfer to mouse recipients with mouse embryos. The mouse day7 and day 9 embryos implantation sites were use as control. Experiment Overall Design: totally 6 samples were analyzed, each samples two replications (one of them had three replications).

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Silencing or amplification of endocannabinoid signaling in blastocysts via CB1 compromises trophoblast cell migration

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