Alterations of RNA Modification in Mouse Germ Cell-2 Spermatids Under Hypoxic Stress

Abstract

Hypoxia is a known stress factor in mammals and has been shown to potentially impair male fertility, which manifests as spermatogenic dysfunction and decreased semen quality. Studies have shown that RNA modifications, the novel post-transcriptional regulators, are involved in spermatogenesis, and hypoxia-induced alterations in RNA modification in testes and sperm cells may be associated with impaired spermatogenesis in mice. However, the molecular mechanisms via which RNA modifications influence spermatogenesis under hypoxic stress conditions are unclear. In this study, we generated a mouse Germ Cell-2 spermatid (GC-2spd) hypoxia model by culturing cells in a 1% O2 incubator for 48 h or treating them with CoCl2 for 24 h. The hypoxia treatment significantly inhibited proliferation and induced apoptosis in GC-2spd cells. The RNA modification signatures of total RNAs (2 types) and differentially sized RNA fragments (7 types of approximately 80 nt-sized tRNAs; 9 types of 17–50 nt-sized sncRNAs) were altered, and tRNA stability was partially affected. Moreover, the expression profiles of sncRNAs, such as microRNAs, tsRNAs, rsRNAs, and ysRNAs, were significantly regulated, and this might be related to the alterations in RNA modification and subsequent transcriptomic changes. We comprehensively analyzed alterations in RNA modification signatures in total RNAs, tRNAs (approximately 80 nt), and small RNAs (17–50 nt) as well as the expression profiles of sncRNAs and transcriptomes in hypoxia-treated GC-2spd cells; our data suggested that RNA modifications may be involved in cellular responses under hypoxic stress conditions and could provide a basis for a better understanding of the molecular mechanisms underlying male infertility.

Cascading effects of hypobaric hypoxia on the testis: insights from a single-cell RNA sequencing analysis

“…The quantity of KIF2C within the sperm is closely associated with fertilization and the early stages of embryogenesis. Animal experiments demonstrated the importance of KIF2C in spermatic mitosis and meiosis ( He T. et al, 2022 ). We found that environmental stimuli upregulate KIF2C in elongated spermatids, thereby influencing spermatic mitosis and normal sperm cell development.…”

Altered landscape of total RNA, tRNA and sncRNA modifications in the liver and spleen of mice infected by Toxoplasma gondii

“…Northern blotting was performed to evaluate tRNA expression levels in the mouse liver during T. gondii infection as previously described [ 29 ]. Total RNA was separated on a 15% urea-PAGE gel.…”

Biological function and clinical application prospect of tsRNAs in digestive system biology and pathology

“…(4) Although tsRNAs exist in serum, plasma, saliva, and urine, tsRNAs themselves have many modifications [ 135 ]. The stability of tsRNAs, their capacity to be recognized through RNase, and ultimately their ability to generate tsRNAs are all impacted by these alterations, which considerably affect the structure and function of tsRNAs [ 136 ]. Therefore, whether the different types of tsRNAs produced at different sites can be stably expressed in exosomes as biomarkers still needs to be verified in many investigations.…”