miR-199-5p regulates spermiogenesis at the posttranscriptional level via targeting Tekt1 in allotriploid crucian carp

Abstract

Background Sperm abnormalities are one of the primary factors leading to male sterility, but their pathogenesis is still unclear. Although miRNAs are suggested to exert important roles in the regulation of spermatogenesis at both transcriptional and posttranscriptional levels, little is currently known regarding the regulation of sperm flagella assembly by microRNAs (miRNAs). The role of miRNAs in the development of sperm abnormalities in sterile triploid fish has not been studied. Results In this study, we found that miR-199-5p was widely expressed in all detected tissues of different-ploidy crucian carp. As one of the testis-specific candidate markers, Tekt1 was predominantly expressed in the testis. Quantitative real-time PCR (qRT-PCR) analyses showed that the expression trend of miR-199-5p was exactly opposite to that of Tekt1. Through bioinformatics analysis, we identified a putative miR-199-5p binding site in the Tekt1 mRNA. We further identified Tekt1 as a target of miR-199-5p using luciferase reporter assay. Finally, we confirmed that miR-199-5p was necessary for sperm flagellar assembly and spermatogenesis in vivo via intraperitoneal injection of miR-199-5p antagomir or agomir in diploid red crucian carp. Moreover, miR-199-5p gain-of-function could lead to spermatids apoptosis and abnormal spermatozoa structure, which is similar to that of allotriploid crucian carp. Conclusions Our studies suggested that abnormally elevated miR-199-5p inhibited the sperm flagella formation in spermiogenesis by negatively regulating the expression of Tekt1, thereby causing sperm abnormalities of male allotriploid crucian carp.

Glucocorticoid exposure modifies the miRNA profile of sperm in the guinea pig: Implications for intergenerational transmission

“…Specifically, miR‐199, miR‐449a and miR‐125 were significantly decreased in cauda sperm (by microarray analysis). Previous studies have demonstrated that miR‐199 is important for regulating spermiogenesis by binding to the 3’‐UTR of one of its target gene, Tekt1 47 . Tekt1 has been demonstrated to influence the production and maturation of sperm 47,48 .…”

“…47 Tekt1 has been demonstrated to influence the production and maturation of sperm. 47,48 Other studies have demonstrated that reduced miR-199 levels in IVF blastocysts decreases their viability and developmental potential. 49 miR-449 has been demonstrated to be significantly reduced in sperm of mice exposed to chronic stress and these changes persist to the early embryo.…”

“…Previous studies have demonstrated that miR‐199 is important for regulating spermiogenesis by binding to the 3’‐UTR of one of its target gene, Tekt1 47 . Tekt1 has been demonstrated to influence the production and maturation of sperm 47,48 . Other studies have demonstrated that reduced miR‐199 levels in IVF blastocysts decreases their viability and developmental potential 49 .…”

Alternative signal pathways underly fertilization and egg activation in a fish with contrasting modes of spawning

“…Examination of ultrastructure also showed that the vestibule of adhesive eggs was recessed and wider than that of semi-buoyant eggs, which appears adaptive in adhesive eggs in facilitating sperm entry to attached eggs, in contrast with semi-buoyant eggs in which mixing of eggs and sperm during mating likely gives greater assurance of fertilization. However, the guidance mechanisms for sperm entry during fertilization in fishes remains relatively poorly understood [ 45 , 46 ], and the adaptive significance, if any, of the contrasting forms of vestibule between adhesive and semi-buoyant eggs of C. alburnus remains to be demonstrated.…”

Kaempferol Promotes Apoptosis and Inhibited Autophagy in A549 Cells via MicroRNA-199/Mammalian Target of Rapamycin Axis

“…Kaempferol can inhibit the proliferation and increases apoptosis in cancer cells by triggering apoptosis, cell cycle and downregulation of signaling pathways [10,[12][13][14] . Micro Ribonucleic Acid (RNA), as a small non-coding RNA, can regulate the RNA expression [15][16][17] . Previous study shows that microRNA (miR)-199 was low expressed in many cancers such as liver cancer, lung cancer etc.…”