Next-generation sequencing-based microRNA profiling of mice testis subjected to transient heat stress
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Meng Rao1,*, Zhengyan Zeng2,*, Li Tang1, Guiping Cheng3, Wei Xia3 and Changhong Zhu3
1Department of Reproduction and Genetics, The First Affiliated Hospital of Kunming Medical University, Kunming, China
2Department of Neurology, The First Affiliated Hospital of Kunming Medical University, Kunming, China
3Family Planning Research Institute, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, China
*These authors have contributed equally to this work
Meng Rao, email: email@example.com
Keywords: heat stress; spermatogenesis; microRNA; next-generation sequencing; apoptosis
Received: October 07, 2017 Accepted: November 16, 2017 Published: December 04, 2017
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.
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