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Transcriptome Profile of Mouse Trophoblast Giant Cells:Effects of Oxycodone on these and other Placental Lineages

ABSTRACT: Pregnant women may be prescribed opioid drugs for pain or may abuse these drugs. The placenta may be key to understanding how opioids cause adverse pregnancy outcomes. Maternal oxycodone (OXY) exposure of pregnant mice leads to disturbances in the layer of invasive parietal trophoblast giant cells (pTGC) that forms the interface between the placenta and uterus and is analogous to extravillous trophoblasts of the human placenta. These cells are probably crucial to coordinating the metabolic needs of the conceptus with those of the mother and are also primary participants in the placenta-brain axis. Their large nuclear size, however, has precluded both single cell (sc) and single nucleus (sn) RNA-seq analyses beyond embryonic day (E) 8.5. Here we compared the transcriptomes of placentas from pregnant mice exposed to OXY with unexposed controls at E12.5, with particular emphasis on the pTGC. The non-fluidic Parse snRNA-seq approach permitted characterization of the nuclear transcriptomes of the major placental cell lineages and their presumed progenitors at E12.5. OXY exposure had negligible effect on components of the placental labyrinth, including the two syncytial cell layers, but caused transcriptomic changes consistent with metabolic stress throughout the spongiotrophoblast. Most notably, there was loss of the majority of pTGCs, whose normal gene expression is consistent with elevated energy demand relating to biosynthesis of multiple secretory products, especially hormones, and endoduplication of DNA. This unusual sensitivity of pTGC may put the pregnancy and future health of the offspring at particular risk to OXY exposure.

ORGANISM(S): Mus musculus

PROVIDER: GSE271203 | GEO | 2024/07/01

REPOSITORIES: GEO

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Project description:Purpose: The goals of this study are to examine how maternal exposure to oxycodone affects the global transcriptome and histological structure of the mouse placenta at 12.5 dpc of pregnancy.retinal transcriptome profiling (RNA-seq) to microarray and quantitative reverse transcription polymerase chain reaction (qRT–PCR) methods and to evaluate protocols for optimal high-throughput data analysis Methods: A high throughput RNA sequencing method was used to examine the global transcriptomic changes affected by oxycodone in male and female conceptuses. Histomorphometrics were used to assess the various regions of the placenta, namely the labyrinth, spongiotrophoblast, and parietal trophoblast giant cell (pTGC) region. The open-sourced software, weighted correlation network analysis (WGCNA) was used to link gene expression changes and histological (phenotypic) changes. Results: Using an optimized data analysis workflow, we mapped over 40 million sequence reads per sample to the mouse genome (build grcm38). An average of 95.6% of reads (38.7 million) was mapped with HISAT2 workflow. We identified 12,138 transcripts in the placenta of saline control and oxycodone mice with featureCounts workflow. One hundred eighty-eight transcripts showed differential expression between the OXY male vs. CTL male placenta, with a log2Foldchange ≥1 and p value <0.05. Fifty-eight transcripts showed differential expression between the OXY female vs. CTL female placenta, with a log2Foldchange ≥1 and p value <0.05. Hierarchical clustering of differentially expressed genes uncovered several genes that may contribute to placenta function. Conclusions: Our study represents the first detailed analysis of placental transcriptomes in oxycodone-exposed mice, with biologic replicates, generated by RNA-seq technology. The data analysis workflows reported here should provide a framework for comparative investigations of expression profiles. We conclude that oxycodone exposure changed transcriptome characterization and these changes were associated with placenta histological features.

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Transcriptome Profile of Mouse Trophoblast Giant Cells:Effects of Oxycodone on these and other Placental Lineages

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