Global gene expression profiling of individual human oocytes and embryos demonstrates heterogeneity in early development lisa shaw1,2,3. For global gene expression profiling of preimplantation embryos, c57bl6j mice aged 45 weeks were superovulated by injections of 5 iu pregnant mare serum gonadotropin pms, sigma, st. Within this period, although there are little morphological differences between the human and mouse embryo, the global gene expression patterns show dissimilarities. In the current study, we investigated whether in vitro culture of preimplantation mouse embryos can affect their subsequent development and can lead to a deregulation of genomic imprinting. Brison1,2 1faculty of medical and human sciences, university of manchester, manchester, united kingdom, 2department of reproductive medicine, old st. In this study, we analyzed the gene expression variability of human embryos at different developmental stages. Gene expression profiling an overview sciencedirect topics.
While this work was being prepared for publication, two studies appeared that used microarrays to assess global patterns of gene expression during preimplantation development hamatani et al. Advances in transgenic and knockout studies in mouse have dramatically strengthened the use of this model and significantly improved our understanding of gene function during development in the past few decades. The scarcity of the materials of preimplantation embryos, both in size diameter global gene expression profiling of preimplantation embryos. Hamatani t1, ko msh, yamada m, kuji n, mizusawa y, shoji m, hada t, asada h, maruyama t, yoshimura y. In the oocyte figure 2a, approximately 16,000 different transcripts were expressed across the three samples i. Culture of preimplantation mouse embryos affects fetal. Human gene expression first occurs between the four and eightcell stages of preimplantation development. Preimplantation development is marked by four major events. Dynamics of global gene expression changes during mouse. To identify the best reference genes for gene expression studies in mouse preimplantation embryos, an rtqpcr assay based on large amounts of transcriptome data from the mammalian preimplantation embryos was designed for the transcription profiling of the ten genes actb, gapdh, sdha, ablim, ywhaz, sptbn, h2afz, tgfb1, 18 s and wrnip. A greater impact on gene expression might have been observed if the embryos had been derived by art followed by assisted reproduction techniques followed by in vitro culture because in vitro fertilization and embryo culture can affect gene expression 15 x 15 rinaudo, p.
Our gene expression profiling during preimplantation development revealed the distinctive patterns of maternal rna degradation and. Global gene expression profiling and senescence biomarker. Global gene expression profiling of individual human. Evaluation of reference genes in mouse preimplantation.
Transcriptional reprogramming of somatic cell nuclei during preimplantation development of cloned bovine embryos. Unsupervised hierarchical clustering showed that global gene expression profiles of individual cells at the same stage, from oocyte. Hence, we determined the effects of embryo culture media with or without serum on pre and postim. Article global gene expression profiling of individual. B representative pearson correlation coefficient for replicates of pig oocyte samples calculated using log2based rpkm. The scatterplot data in figure 1 are presented as venn diagrams in figure 2. Expression profiling of preimplantation embryos a experimental design.
Here, we determined for the first time the transcriptomes of bovine metaphase ii oocytes and all stages of preimplantation embryos. This study was designed to address this question by comparing global gene expression profiles from all the stages of preimplantation development in vitro and in vivo. An integrated chromatin accessibility and transcriptome. Microarrays on oocytes and preimplantation embryos using this specific rna isolation and amplification protocol have allowed us to. Application of this technology to oocytes and preimplantation embryos has been. Transcript profiling during preimplantation mouse development. Singlecell rnaseq profiling of human preimplantation. Profiling gene expression in growtharrested mouse embryos. Survival and cell death signals are crucial for mammalian embryo preimplantation development. We discovered distinctive patterns of variability across cells in the embryo. Little is known about the gene expression pattern in early embryonicpreimplantation development due to the difficulty of material collection and sensitivity of molecular tools used for analysis. After fertilization, a rich supply of maternal proteins and rnas support development whereas a number of zygotic and embryonic genes are expressed in a stagespecific manner leading to embryonic genome activation ega. Transcriptional reprogramming of somatic cell nuclei during preimplantation development of.
Recently, the efforts to characterize cleavage stage embryos have revealed the global gene expression profiles during preimplantation development of mouse embryo 78 9 10111214. T1 a global view of gene expression in the preimplantation mouse embryo. Analysis of transcription factor expression during. The scarcity of the materials of preimplantation embryos, both in size diameter global gene expression profiling to the study of preimplantation embryos. Gene expression landscape of preimplantation embryonic development. Transcript profiling during preimplantation mouse development transcript profiling during preimplantation mouse development zeng, fanyi. Aberrations in gene expression pattern, associated with in vitro methods, such as in vitro maturation ivm, ivf and ivc of embryos have been demonstrated rizos et al. U, f, 2, 4, 8, m, and b denote unfertilized egg, fertilized egg, 2cell embryo, 4cell embryo, 8cell embryo, morula, and blastocyst, respectively. Human preimplantation embryo development development. Blastocyst gene expression correlates with implantation. Analysis of transcription factor expression during oogenesis and preimplantation development in mice volume 15 issue 2 s. Functional studies of ashwin suggest that it participates in cell survival and anteroposterior patterning. Effects of embryo culture on global pattern of gene. Variability of gene expression identifies transcriptional.
What can we learn from gene expression profiling of mouse. The variability in gene expression which we have found between oocytes and embryos at the same stage requires the reinterpretation of previous microarray studies based on pooling a number of. The effect of blastomere biopsy on preimplantation mouse. The aim of this study was therefore to investigate the global gene expression profile at three key developmental stages spanning human preimplantation embryo development. Gene expression profiling using microarray technology is a robust, efficient, and costeffective approach to assay a cell or tissues transcriptome at a particular time or under a specific condition. Animals free fulltext ashwin gene expression profiles. Understanding human preimplantation development has important implications for assisted reproductive technology art and for human embryonic stem cell hescbased therapies. Bovine embryos can be generated by in vitro fertilization or somatic nuclear transfer. Furthermore, ashwin is expressed zygotically in this species, which suggests that it participates in embryonic.
Mouse has served as an excellent model for studying human development and diseases due to its similarity to human. The magnitude of the effect on the global pattern of gene expression, however, is not known. We describe a global analysis of gene expression at stages spanning human preimplantation development, at the level of the individual oocytesembryos. Owing to limited resources, the cellular and molecular mechanisms governing this early stage of human development are poorly understood. This second wave is believed to drive the dramatic morphological changes of compaction and differentiation into the two cell lineages of the blastocyst 5 x 5 hamatani, t. Measuring gene expression in individual cells is crucial for understanding the gene regulatory network controlling human embryonic development.
The results of this study provided the first global analysis of the human preimplantation embryo. Global, survival, and apoptotic transcriptome during mouse. In cats, only few studies have been carried out to examine early embryonic gene expression pattern. Analysis of gene expression profiles in preimplantation. In the past, large numbers of embryos have been needed to get sufficient material for analysis. Global transcriptome analysis of different stages of.
Global gene expression profiling of mouse preimplantation embryos to dissect maternal transcripts. Preimplantation gene expression profiles 119 figure 1. Pdf transcriptional reprogramming of somatic cell nuclei. Mouse studies are widely used to understand preimplantation embryo development, but extrapolation of these results to humans is questionable. Brison1,2 1faculty of medical and human sciences, university of manchester, manchester, united kingdom, 2department of reproductive medicine, old st marys hospital, central.
However, the knowledge on the molecular mechanisms underlying their regulation is still limited. Recent progress in rna amplification methods and microarray platforms, including genes unique to preimplantation embryos, allow us to apply global gene expression profiling to the study of. A the porcine preimplantation embryonic developmental process from left to right. Real time rtpcr with in vitro embryos, black columns.
Differential gene expression analysis identified 3048 upregulated and 2169 downregulated transcripts from mii eggs to 8to16cell embryos fc 2, fdr global activation of the embryonic genome is the most critical event in early mammalian development. A global view of gene expression in the preimplantation. Transcript profiling of bovine embryos implicates specific. Here we propose a novel comparative analysis integrating an already published global gene expression by singlecell rnaseq profiling of individual human oocytes and embryos derived from assisted reproduction procedures art and the global gene expression profiling of hescs exposed to noncytotoxic oxidative stress conditions obtained in this. Two groups simultaneously published the first reports on global gene expression profiling of all stages of preimplantation embryos fig. Culture of preimplantation embryos affects gene expression. Factors affecting the gene expression of in vitro cultured. Global expression profiling reveals genetic programs. This study examined the gene expression profiles of mouse embryos cultured in vitro, including eight different stages. Transcriptomic profiling, either globally level or at the singlecell level, has been applied to the study ped in model animals mouse and bovine and in. Gene expression profiling of human oocytes developed and. Global gene expression profiles of individual oocytes and blastocysts. The time course of events leading to embryo implantation in the mouse.
Global gene expression profiling of preimplantation embryos. In addition, tedious, laborintensive molecular analysis such. Based on the number of differentially expressed genes degs, developmental stage 3519 degs and maternal age 1258 degs had a larger effect on the global gene expression profile of human preimplantation embryos than either tested culture medium 596 degs or oxygen concentration 492 degs used during in vitro culture. The ashwin gene, originally identified in xenopus laevis, was found to be expressed first in the neural plate and later in the embryonic brain, eyes, and spinal cord. Real time rtpcr with in vivo derived embryos, shaded columns. Recent progress in rna amplification methods and microarray platforms, including genes unique to preimplantation embryos, allow us to apply global gene expression profiling to the study of preimplantation embryos.