Prenatal and postnatal exposure to acetaminophen and mouse neurodevelopment

Abstract pending Pregnant mice (n=12/group) were randomly assigned to receive either acetaminophen (150 mg/kg/day) or no acetaminophen (vehicle control) via palatable gelatin tablets, once a day from embryonic day 10 through postnatal day 21 (weening). At birth, 12 offspring per group, half male and half female, were removed from their litters for tissue harvesting. Prefrontal corticies were harvested, and RNA was extracted with Qigen RNeasy kits. RNA sequencing was performed with the Illumina HiSeq 4000 platform. FASTQ files were preprocessed with fastp using the default settings to filter bad reads, trim low quality bases, and cut adaptors. (https://academic.oup.com/bioinformatics/article/34/17/i884/5093234). Reads were mapped to the GRCM38(mm10) mouse genome using BWA-MEM (https://arxiv.org/abs/1303.3997) and assigned to exons using featureCounts (https://academic.oup.com/bioinformatics/article/30/7/923/232889) before performing differential expression analysis using DESeq2 (https://genomebiology.biomedcentral.com/articles/10.1186/s13059-014-0550-8). Gene expression fold-changes compared acetaminophen treated to control offspring, accounting for sex. Normalized counts from DESeq2 were input into EGSEA (https://academic.oup.com/bioinformatics/article/33/3/414/2875813) to determine enrichment for MSigDB hallmark gene sets (https://www.pnas.org/content/102/43/15545.abstract) (https://www.cell.com/fulltext/S2405-4712(15)00218-5) and KEGG Pathways (https://scholar.google.com/scholar?hl=en&as_sdt=0%2C33&q=KEGG%3A+integrating+viruses+and+cellular+organisms&btnG=). We examined the top 20 EGSEA enrichments sorted by median ranking score and considered pathways significant if FDR-adjusted P<0.05.