Revealing RNA poly(A) tail dynamics during spermiogenesis and its function to support male fertility [dRNA-seq_wildtype]

Poly(A) tail processing of mRNAs have been shown to dictate mRNAs metabolism of a few transcripts critical for fertility, but the extent of poly(A) tail-mediated regulation at the transcriptome level remains poorly characterized. Here, we performed whole transcriptome poly(A) tail profiling of purified late spermatocytes and round spermatids using direct RNA sequencing. As the germ cells enter spermiogenesis, we show a global increase in poly(A) tail length up to ~150-nts associated with transcript stabilization. Overall design: Pachytene spermatocytes and round spermatids were isolated from adult mice testis using Hoescht/PI staining and FACS sorting. Cells were isolated from C57BL/6J/B6 male mice. Each replicate is representative of a pool of 3 individual mice - 3 replicates per condition were considered for analysis. Nanopore technologies were used to sequence the testicular transcriptome and determine mRNA poly(A) tail length.