Original analysis results of GSEA and AS in human spermatozoa

The Gene Set Enrichment Analysis (GSEA) and Alternative Splicing (AS) events were found in RNA-sequencing data for human spermatozoa. This study aimed to compare the unknown micro epigenetics alternations in fresh and cryopreserved spermatozoa for evaluation of the effectivity of respective cryopreservation protocols. Materials and Methods Spermatozoa were divided into three groups: fresh spermatozoa (Group 1), cryoprotectant-free vitrified spermatozoa (Group 2), and conventionally frozen spermatozoa (Group 3). Nine RNA samples (3 replicates in each group) were detected, and they were applied for library preparation using Illumina compatible kit and sequencing by the Illumina platform. Results There are three Gene Ontology (GO) terms that have been enriched: Mitochondrial tRNA aminoacylation, ATP dependent microtubule motor activity, and male meiotic nuclear division in vitrified spermatozoa in contrast with fresh spermatozoa. For Alternative Splicing (AS) analysis, it was found a number of unknown AS events. Including functional genes Extron Skipping (ES), alternative 5′ splice sites (A5SS), alternative 3′ splice sites (A3SS), mutually exclusive Extron (MXE), and retained intron (RI).