miRNA Expression Profiles of Mouse Round Spermatids in GRTH/DDX25-Mediated Spermiogenesis: mRNA–miRNA Network Analysis

Abstract

GRTH/DDX25 is a testis-specific DEAD-box family of RNA helicase, which plays an essential role in spermatogenesis and male fertility. There are two forms of GRTH, a 56 kDa non-phosphorylated form and a 61 kDa phosphorylated form (pGRTH). GRTH-KO and GRTH Knock-In (KI) mice with R242H mutation (lack pGRTH) are sterile with a spermatogenic arrest at step 8 of spermiogenesis due to failure of round spermatids (RS) to elongate. We performed mRNA-seq and miRNA-seq analysis on RS of WT, KI, and KO to identify crucial microRNAs (miRNAs) and mRNAs during RS development by establishing a miRNA–mRNA network. We identified increased levels of miRNAs such as miR146, miR122a, miR26a, miR27a, miR150, miR196a, and miR328 that are relevant to spermatogenesis. mRNA–miRNA target analysis on these DE-miRNAs and DE-mRNAs revealed miRNA target genes involved in ubiquitination process (Ube2k, Rnf138, Spata3), RS differentiation, and chromatin remodeling/compaction (Tnp1/2, Prm1/2/3, Tssk3/6), reversible protein phosphorylation (Pim1, Hipk1, Csnk1g2, Prkcq, Ppp2r5a), and acrosome stability (Pdzd8). Post-transcriptional and translational regulation of some of these germ-cell-specific mRNAs by miRNA-regulated translation arrest and/or decay may lead to spermatogenic arrest in KO and KI mice. Our studies demonstrate the importance of pGRTH in the chromatin compaction and remodeling process, which mediates the differentiation of RS into elongated spermatids through miRNA–mRNA interactions.

Identification of runs of homozygosity associated with male fertility in Italian Brown Swiss cattle

“…Recent research suggests that LIAS may play a critical role in spermatogenesis, as it was shown to be downregulated in cases of spermatogenic dysfunction in humans ( Zhao et al, 2023 ). The UBE2K gene encodes a protein that is a member of the ubiquitin-conjugating enzyme family and is crucial for the ubiquitination pathways, which are essential to the later stages of spermatid development during spermiogenesis ( Anbazhagan et al, 2023 ). Malfunctions in the ubiquitination system can impair sperm development, ultimately resulting in male infertility ( Nakamura, 2013 ).…”

MicroRNAs in spermatogenesis dysfunction and male infertility: clinical phenotypes, mechanisms and potential diagnostic biomarkers

“…miR-34bc - / - ;449 - / - mice exhibit a decrease in the proportion of spermatozoa, elongating spermatids, and abnormalities in sperm morphology with the separation of spermatozoa heads and tails ( 11 ). In one study, miR-32, miR-184, miR-335, miR-140, miR-141, miR-1981, miR-202, miR-880, and miR-669c were downregulated in both KO and KI mice, whereas miR-150, miR-196a-2, miR-652, miR-146, miR-10b, miR-379, miR-122a, miR-26a, miR-27a, miR-127, and miR-328 were upregulated in both KO and KI mice and negatively regulated spermatid differentiation ( 117 ). Liu et al.…”

Small Non-Coding RNAs in Male Reproduction

“…59 DDX25 was also shown to affect the expression levels of other miRNAs in addition to miR-469; therefore, it appears to have a broader role in the control of miRNA-mRNA networks to support haploid differentiation of spermatids. 60 Another miRNA, miR-122a, that is expressed in late stages of male germ cells differentiation has also been shown to directly control TNP2 expression by inducing mRNA cleavage. 61…”