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• W4387396235 abstract "Single cell analysis of embryos has its limitations, since the specific positioning of cells within the embryo is unknown. Our aim was to use a novel in situ gene expression technology to analyze the expression of up to 80 genes per cell in the human blastocyst and three-dimensionally map these genes within each cell to gain a better understanding of their specific location. We performed targeted in situ RNA sequencing of 80 genes in 15 human blastocysts using sequential rounds of RNA in situ sequencing. Specific RNA transcripts were hybridized then amplified for the chosen genes. The read out was then done over several rounds of in situ sequencing. With each round, the transcript was labelled, and the entire volume of the embryo was imaged. This technique was first performed on mouse blastocysts and the spatial distribution of ∼150 genes was charted in mouse embryos using publicly available single cell RNAseq data sets of preimplantation mouse embryos. Similarly, the publicly available RNAseq data of human blastocysts was used to construct similar blastocysts lineage markers. We performed targeted in situ RNA sequencing of 80 genes in 15 human blastocysts and distinguished three distinct cell types: trophectoderm (TE), Epiblast (Epi), and Primitive Endoderm (PrE). Using the high multiplexing capacity of the 80-gene panel, we identified subpopulations and specific positioning of Epi and TE, including polar TE (pTE) and mural TE (mTE), and discovered novel gene markers expressed explicitly in these populations. By investigating the gene expression patterns within these subpopulations, we identified gene regulatory programs specific to the Epi and each TE subtype, providing deeper insights into the molecular mechanisms underlying Epi and TE differentiation and development in human blastocysts. In addition, we compared our findings in human blastocysts to mouse embryos. Through this comparison, we assessed cell type programs' expression and spatial localization and identified conserved and divergent gene expression patterns between human and mouse embryos. Furthermore, we compared the gene regulatory modules in polar and mural TE and found differences in the expression of genes related to cell adhesion and differentiation. Our study demonstrates the power of targeted in situ RNA sequencing to identify and localize the expression of multiple genes simultaneously within the human blastocyst. By using a high multiplexing capacity of 80 genes, we were able to identify subpopulations of Epi and TE and define gene regulatory programs specific to each subtype. Our results provide a comprehensive view of gene expression dynamics during early human development and could aid in the understanding of the molecular mechanisms underlying Epi and TE differentiation and development. Overall, our approach could be applied to other tissues and developmental stages, providing a valuable resource for future studies on gene expression during development and disease." @default.
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• W4387396235 date "2023-10-01" @default.
• W4387396235 modified "2023-10-07" @default.
• W4387396235 title "USING IN SITU GENE EXPRESSION TO CREATE A THREE-DIMENSIONAL MAP OF HUMAN BLASTOCYSTS" @default.
• W4387396235 doi "https://doi.org/10.1016/j.fertnstert.2023.08.744" @default.
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