MicroRNA-targeting in male infertility: Sperm microRNA-19a/b-3p and its spermatogenesis related transcripts content in men with oligoasthenozoospermia

Abstract

Objective: To elucidate and validate the potential regulatory function of miR-19a/b-3p and its spermatogenesis-related transcripts content in sperm samples collected from men with oligoasthenozoospermia.Methods: Men presenting at an infertility clinic were enrolled. MicroRNA (miRNA) and target genes evaluation were carried out using in silico prediction analysis, Reverse transcription-quantitative PCR (RT-qPCR) validation, and Western blot confirmation.Results: The expression levels of miRNA-19a/b-3p were significantly up-regulated and 51 target genes were significantly down-regulated in oligoasthenozoospermic men compared with age-matched normozoospermic men as determined by RT-qPCR. Correlation analysis highlighted that sperm count, motility, and morphology were negatively correlated with miRNA-19a/b-3p and positively correlated with the lower expression level of 51 significantly identified target genes. Furthermore, an inverse correlation between higher expression levels of miRNA-19a/b-3p and lower expression levels of 51 target genes was observed. Consistent with the results of the RT-qPCR, reduced expression levels of STK33 and DNAI1 protein levels were identified in an independent cohort of sperm samples collected from men with oligoasthenozoospermia.Conclusion: Findings suggest that the higher expression of miRNA-19a/b-3p or the lower expression of target genes are associated with oligoasthenozoospermia and male infertility, probably through influencing basic semen parameters. This study lay the groundwork for future studies focused on investigating therapies for male infertility.

A comprehensive analysis of spermatozoal RNA elements in idiopathic infertile males undergoing fertility treatment

“…Hsa-miR-224, which was also enriched in the low blastocyst rate group, has been functionally-related to the differentiation of spermatogonial stem cells in mice 51 . Higher expression of miRNA-19a/b-3p has previously been associated with oligoasthenozoospermia and male infertility 52 . While miRNA-19a/b-3p was indeed down-regulated in the high blastocyst rate group, it was also reduced in the low group, suggesting its activity in normozoospermic males may require further elucidation.…”

MiR-19b-3p inhibits cell viability and proliferation and promotes apoptosis by targeting IGF1 in KGN cells

“…It has been demonstrated that miR-19b is involved in the regulation of various biological process, including cellular proliferation, differentiation, migration, invasion and apoptosis. It was reported that higher expression of miR-19b-3p were associated with oligoasthenozoospermia and male infertility (30). MiR-19b-3p was differentially expressed in the spent culture media after embryo transfer and showed an AUC value for predicting positive pregnancy outcomes, both, from spent culture media and sperm collected from infertile couples attending infertility treatment (31).…”

Sperm Motility Annotated Genes: Are They Associated with Impaired Fecundity?

“…Complementary DNA (cDNA) was generated in 5 µL reactions by reverse transcription (RT) of 75 ng RNA using the Reverse Transcription Master Mix (Fluidigm Corporation, South San Francisco, CA, USA), as previously described [17]. After the RT reaction was completed, the reactions were used immediately for preamplification reactions with Preamp Master Mix (Fluidigm) according to the manufacturer's protocol PN 100-5875 C1.…”

“…Biomark ™ HD with a 96.96 IFC was used for the RT-qPCR amplification, as previously described [17]. Briefly, for each sample, a 6 µL sample mix containing 3 µL of 2× SsoFast EvaGreen Supermix with low ROX (Bio-Rad Laboratories, Hercules, CA, USA), 0.3 µL of 20× DNA Binding Dye (Fluidigm), and 2.7 µL of the pre-amplified sample was prepared.…”