Deep Sequencing Analysis of microRNAs in Bovine Sperm

microRNAs (miRNAs) are small non-coding RNAs that participates in the regulation of many physiological pathways, but a role for spermatozoon-delivered miRNAs in fertilization and embryonic development remains controversial. A library of miRNAs in bovine sperm was constructed using Illumina high-throughput sequencing technology, along with the predication and the pathway analysis of target genes. miRNAs in mammalian spermatozoon were systematically investigated, and a protocol for RNA isolation from the cauda region of an epididymal biopsy was established. Unique sequences that were 18-26 nucleotides in length were mapped to specific precursors in miRBase 20.0 using BLAST. A total of 951 known miRNAs and 8 novel, highly expressed miRNA candidates were identified. The search for endogenous sperm miRNAs will contribute to a preliminary database for functional and molecular mechanistic studies in embryonic development and sperm epigenetic programming. Mol. Reprod. Dev. 81: 1042-1052, 2014. (c) 2014 Wiley Periodicals, Inc.