P-745 Follicular fluid exposure may activate gene expression in mature human sperm before fertilization

Abstract Study question Does follicular fluid (FF) exposure shape human sperm RNA content and are these changes acquired through genomic or nongenomic manner? Summary answer FF altered potentially via activation of transcription machinery caused selective intake transcripts in sperm. What is known already Mature have been widely recognized as transcriptionally inert. Therefore, RNAs regarded the sole remnants gene expression process that occurs during spermatogenesis. Supporting this view, most histones replaced by protamines developing spermatids to enable tighter packaging genetic material, which should prevent mature cells express their genes. However, humans, approximately 15% DNA remains associated with histones, raising possibility genome regions could be available for de novo under an appropriate stimulus. design, size, duration Fresh, were divided into three aliquots. One aliquot was used untreated control sample, second third aliquots treated either PureSperm® Wash (SW) solution pool five females hours. sequencing (RNA-seq) data collected from samples. Results complemented Chromatin Immunoprecipitation Sequencing (ChIP-seq) (GEO: GSE15690 GSE49624) nascent capture followed PCR. Participants/materials, setting, methods All male female participants reproductive age, non-smoking, healthy individuals who reported no history infertility. Differentially expressed genes (DEG) identified based on RNA-seq (adjusted p-value < 0.05, logFC |0.75|). The histone status loci DEGs evaluated using ChIP-seq data. also subjected Gene Ontology analysis. Activated confirmed Click-iT™ Nascent Capture kit Main results role chance showed 193 between FF-treated samples 30 upregulated 163 downregulated. Furthermore, 18 detected SW- 17 one Finally, 137 (all downregulated) found SW-treated Among genes, chemokine-signaling, ion transport, steroidogenesis-related biological functions enriched whereas processing, translation, protein targeting-related processes among downregulated (untreated vs. FF-treated). revealed 93% located within histone-bound chromatin them bear both active inactive markers. we upregulation at least two (CXCL3 CXCL8) resulted sperm, IGF2 due internalization mRNA FF. Additionally, 1396 protein-coding mRNAs present expression-related such regulation transcription, processing subtypes, translation. Limitations, reasons caution Due limited sample generalizability our validated future. molecular mechanisms behind observed production needs clarified more detail, including potential FF-mediated remodeling markers involved expression. Wider implications findings Our demonstrate content, seems partly has believed non-existent. can many important a deeper understanding fertilization, early embryogenesis, Trial registration number Not applicaple