Transcriptome profiling of individual rhesus macaque oocytes and preimplantation embryos†

Abstract

Early mammalian embryonic transcriptomes are dynamic throughout the process of preimplantation development. Cataloging of primate transcriptomics during early development has been accomplished in humans, but global characterization of transcripts is lacking in the rhesus macaque: a key model for human reproductive processes. We report here the systematic classification of individual macaque transcriptomes using RNA-Seq technology from the germinal vesicle stage oocyte through the blastocyst stage embryo. Major differences in gene expression were found between sequential stages, with the 4- to 8-cell stages showing the highest level of differential gene expression. Analysis of putative transcription factor binding sites also revealed a striking increase in key regulatory factors in 8-cell embryos, indicating a strong likelihood of embryonic genome activation occurring at this stage. Furthermore, clustering analyses of gene co-expression throughout this period resulted in distinct groups of transcripts significantly associated to the different embryo stages assayed. The sequence data provided here along with characterizations of major regulatory transcript groups present a comprehensive atlas of polyadenylated transcripts that serves as a useful resource for comparative studies of preimplantation development in humans and other species.

TRPM7 and Ca V 3.2 channels mediate Ca 2+ influx required for egg activation at fertilization

“…Although the current studies were performed in the mouse, TRPM7, CACNA1H, and TRPV3 are all expressed in human oocytes and TRPM7 and TRPV3 are expressed in macaque oocytes as indicated by RNA sequencing (45)(46)(47)(48). Furthermore, there are numerous missense or loss of function (LoF) mutations identified in human TRPM7 (480 missense, 27 LoF), CACNA1H (1,045 missense, 10 LoF), and TRPV3 (312 missense, 23 LoF) (see Exome Aggregation Consortium browser at exac.broadinstitute.org; ref.…”

Single-cell RNA sequencing of oocytes from ovarian endometriosis patients reveals a differential transcriptomic profile associated with lower quality

“…Global transcriptomics techniques, such as RNA sequencing (RNA-seq), have recently allowed researchers to determine the whole transcriptome even from single cells (Hwang et al, 2018). RNAseq has already successfully been applied to macaque (Chitwood et al, 2017) and human oocytes (Zhao et al, 2019). However, no previous research has investigated the complete transcriptomic profile of oocytes from endometriosis patients compared to healthy controls.…”

Using Single Molecule mRNA Fluorescent in Situ Hybridization (RNA-FISH) to Quantify mRNAs in Individual Murine Oocytes and Embryos

“…The development of single cell RNA sequencing has overcome some of these limitations, although it still requires linear amplification of cDNA prior to sequencing. It should be noted that this technique has been successfully used to identify changes in mRNA abundance in rhesus macaque, bovine, mouse, and human oocytes and/or embryos 15 – 19 . One-step RT-PCR assays have also been developed but a NCBI search showed that the majority of published data used this technique to primarily detect viral load in mammalian samples.…”