Project description:Embryonic portion of day 6.5 (E6.5) pre-streak mouse embryos. The uterus was removed from 6-day pregnant dams at 9:00 am. 5 females yielded a total of 46 embryos. We discarded embryos with signs of primitive streak or dissection damage. The remaining 38 embryos were dissected to discard the extraembryonic portion and combined to ensure sufficient number for snATACSeq Chromium pipeline.
Project description:Establishment of the mammalian body plan occurs shortly after the embryo implants into the maternal uterus, and our understanding of post-implantation developmental processes remains limited. While methods for in vitro culture of pre- and peri-implantation mouse embryos are routinely utilized, approaches for robust culture of post-implantation embryos from egg cylinder stages until advanced organogenesis remain to be established. We develop herein highly stable ex utero post-implantation mouse embryo culture platforms, that enable appropriate development of embryos before gastrulation (E5.5) until the hind limb formation stage (E11). Late gastrulating embryos (E7.5) are grown in 3D rotating bottles settings, while extended culture from pre-gastrulation stages (E5.5 or E6.5) requires a combination of novel static and rotating bottle culture protocols. Histological, molecular, and single cell RNA-seq analysis validate that the ex utero developed embryos recapitulate precisely in utero development. This culture system is amenable to introducing a variety of embryonic perturbations and micro-manipulations that can be followed ex utero for up to 6 days. Establishment of a system to robustly grow normal mouse embryos ex utero from pre-gastrulation to advanced organogenesis represents a valuable tool to investigate post-implantation embryogenesis, eliminating the uterine barrier to mechanistically interrogate morphogenesis and tissue specification in mammals.
Project description:Regionalized lineage progenitor cells emerged from the onset of gastrulation would facilitate mouse embryonic patterning and build a blueprint for future development. However, the molecular mechanism especially the epigentic mechanism underlying the formation and development of lineage restricted cells has not been fully unveiled. Here,we present a comprehensive landscape of H3K4me3, H3K4me1, H3K9ac, H3K27ac, H3K27me3 and DNA methylation pattern in mouse embryo from pre-streak stage to late-streak stage, and fully analyze the epigenetic mechansim driving the embryonic patterning of mouse gastrula
Project description:The early stages of development of the chick embryo, leading to primitive streak formation (the start of gastrulation), have received renewed attention recently, especially for studies of the mechanisms of large-scale cell movements and those that position the primitive streak in the radial blastodisc. Over the long history of chick embryology, the terminology used to define different regions has been changing, making it difficult to relate studies to each other. To resolve this objectively requires precise definitions of the regions based on anatomical and functional criteria, along with a systematic molecular map that can be compared directly to the functional anatomy. Here, we undertake these tasks. We describe the characteristic cell morphologies (using scanning electron microscopy and immunocytochemistry for cell polarity markers) in different regions and at successive stages. RNAseq was performed for 12 regions of the blastodisc, from which a set of putative regional markers was selected. These were studied in detail by in situ hybridization. Together this provides a comprehensive resource allowing the community to define the regions unambiguously and objectively. In addition to helping with future experimental design and interpretation, this resource will also be useful for evolutionary comparisons between different vertebrate species.
Project description:This SuperSeries is composed of the following subset Series: GSE20974: Bovine pre-transfer endometrium and embryo transcriptome fingerprints as predictors of pregnancy success after embryo transfer (endometrial study) GSE21047: Bovine pre-transfer endometrium and embryo transcriptome fingerprints as predictors of pregnancy success after embryo transfer (embryo study) Refer to individual Series