Title: Next-generation sequencing-based microRNA profiling of mice testis subjected to transient heat stress
Abstract: // Meng Rao 1, * , Zhengyan Zeng 2, * , Li Tang 1 , Guiping Cheng 3 , Wei Xia 3 and Changhong Zhu 3 1 Department of Reproduction and Genetics, The First Affiliated Hospital of Kunming Medical University, Kunming, China 2 Department of Neurology, The First Affiliated Hospital of Kunming Medical University, Kunming, China 3 Family Planning Research Institute, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China * These authors have contributed equally to this work Correspondence to: Meng Rao, email: [email protected] Keywords: heat stress; spermatogenesis; microRNA; next-generation sequencing; apoptosis Received: October 07, 2017     Accepted: November 16, 2017     Published: December 04, 2017 ABSTRACT This study aimed to investigate the role of microRNA (miRNA) in heat stress-induced spermatogenic impairment. Testes from 15 adult ICR mice subjected to testicular hyperthermia at 43°C for 30 min and from 15 control mice were collected and pooled into 3 samples. Isolated RNA from these samples was subjected to small RNA high-throughput sequencing, and differentially expressed miRNAs were identified and validated using RT-PCR. The identified miRNAs were further subjected to Gene Ontology and KEGG analyses, which revealed significant enrichment for pathways potentially involved in heat stress-induced spermatogenic impairment. Additionally, a correlation analysis of the relative levels of validated miRNAs with germ cell apoptosis was performed. Of the 11 miRNAs identified as differentially expressed, 8 were validated as consistent with sequencing data. Further analyses suggested that the target genes of those miRNAs were involved in various pathways (e.g., ribosomal, HIF-1, MAPK) that may be critical to heat stress-induced testicular damage. Some identified miRNAs, including miR-449a-3p, miR-92a-1-5p, miR-423-3p, and miR-128-3p, correlated closely with germ cell apoptosis. The study results reveal a detailed miRNA profile of heat stress-induced testicular damage and highlight new and potentially important candidate targets in the process of male infertility.