Phthalate exposure to adult mice alters non-coding RNAs in cauda epididymosomes

Extracellular vesicles (EVs) are known to contain important cargo of biologically active and regulatory molecules including small non-coding RNA (sncRNA). EVs from cauda epididymis play crucial role in post-testicular maturation of sperm. We assessed levels of cauda epididymosomes sncRNAs following exposure to low dose anti-androgenic Di-n-butyl phthalate (DBP), Di(2-ethylhexyl) phthalate (DEHP), and their mixtures in mice. Overall design: Male mice (C57BL/6J) were treated with vehicle (50% DMSO, n = 6), 2.5 mg/kg/day of DBP (n = 6) or DEHP (n = 6), and phthalate mixture (2.5 mg/kg/day each of DBP and DEHP, n = 6) for six weeks. Cauda epididymosomal RNA were extracted and sequenced to analyze their non-coding RNA (ncRNA) content. Mapping of reads to transcriptomes followed the order: microRNA (miRNA) > transfer RNA fragments (tRFs) > piwi-interacting RNA (piRNA) > ribosomal RNA (rRNA) > circular RNA (circRNA) > long non-coding RNA (lncRNA) using STAR alignment tool. Differential expression of normalized read counts between the exposed groups relative to controls was conducted using EdgeR (Fold change, FC = 1.5 and p < 0.01).