Identification and Functional Analysis of MiRNAs in Extracellular Vesicles of Semen Plasma from High- and Low-Fertility Boars

Overview

Journal Animals (Basel)

Date 2025 Jan 11

PMID 39794983

Authors

Weidong Chen

Yanshe Xie

Zhiqian Xu

Yijun Shang

Wenzheng Yang

Pengyao Wang

Zhenfang Wu

Gengyuan Cai

Linjun Hong

Affiliations

Soon will be listed here.

Abstract

Artificial insemination (AI), as an efficient assisted reproduction technology, can help the livestock industry to improve livestock and poultry breeds, optimize production performance and improve reproductive efficiency. AI technology has been widely used in pig production in China, but boar fertility affects the effectiveness of AI, and more and more studies have shown that there are significant differences in the fertility of boars with similar semen quality indicators. Therefore, this study aimed to identify biomarker molecules that indicate the level of boar fertility, which is important for improving the efficiency of AI. In this study, we collected 40 mL of ejaculates per boar used for extracellular vesicle (EV) characterization in 20 boars and identified 53 differentially expressed miRNAs by small RNA sequencing, of which 44 miRNAs were up-regulated in the high-fertility seminal EVs compared with low-fertility seminal EVs, and nine miRNAs were down-regulated. miR-26a was most significantly down-regulated in the high-fertility group compared to the low-fertility group, and it was hypothesized that this miRNA could be used as a biomolecular marker of semen reproductive performance. To further determine the effect of miR-26a on sperm function, we successfully established a miR-26a overexpression model and found that miR-26a reduced sperm viability, motility, acrosome integrity, plasma membrane integrity and ATP levels. Bioinformatics analysis and dual luciferase reporter analysis revealed that miR-26a directly targets High mobility group A1 (HMGA1). In conclusion, miR-26a can be used as a biomarker to identify high and low fertility in boar semen.

References

Akade E, Jalilian S . The role of high mobility group AT-hook 1 in viral infections: Implications for cancer pathogenesis. Int J Biochem Cell Biol. 2024; 169:106532. DOI: 10.1016/j.biocel.2024.106532. View

Alves M, Celeghini E, Belleannee C . From Sperm Motility to Sperm-Borne microRNA Signatures: New Approaches to Predict Male Fertility Potential. Front Cell Dev Biol. 2020; 8:791. PMC: 7471662. DOI: 10.3389/fcell.2020.00791. View

Wang Y, Zhou C, Meng F, Hu Q, Ding Y, Wang X . Ssc-miR-92b-3p Regulates Porcine Trophoblast Cell Proliferation and Migration via the Gene. Int J Mol Sci. 2022; 23(24). PMC: 9784024. DOI: 10.3390/ijms232416138. View

Knox R . Artificial insemination in pigs today. Theriogenology. 2015; 85(1):83-93. DOI: 10.1016/j.theriogenology.2015.07.009. View

Barranco I, Sanchez-Lopez C, Bucci D, Alvarez-Barrientos A, Rodriguez-Martinez H, Marcilla A . The Proteome of Large or Small Extracellular Vesicles in Pig Seminal Plasma Differs, Defining Sources and Biological Functions. Mol Cell Proteomics. 2023; 22(4):100514. PMC: 10017305. DOI: 10.1016/j.mcpro.2023.100514. View

Zhang L, Liu X, Liu J, Ma X, Zhou Z, Song Y . miR-26a promoted endometrial epithelium cells (EECs) proliferation and induced stromal cells (ESCs) apoptosis via the PTEN-PI3K/AKT pathway in dairy goats. J Cell Physiol. 2017; 233(6):4688-4706. DOI: 10.1002/jcp.26252. View

Hu Q, Zang X, Ding Y, Gu T, Shi J, Li Z . Porcine uterine luminal fluid-derived extracellular vesicles improve conceptus-endometrial interaction during implantation. Theriogenology. 2021; 178:8-17. DOI: 10.1016/j.theriogenology.2021.10.021. View

Battista S, Pentimalli F, Baldassarre G, Fedele M, Fidanza V, Croce C . Loss of Hmga1 gene function affects embryonic stem cell lympho-hematopoietic differentiation. FASEB J. 2003; 17(11):1496-8. DOI: 10.1096/fj.02-0977fje. View

Zeng F, Chen Y, Guo C, Li C, Wei H, Li L . Analysis of differentially abundant proteins related to boar fertility in seminal plasma using iTRAQ-based quantitative proteomics. J Proteomics. 2021; 236:104120. DOI: 10.1016/j.jprot.2021.104120. View

10.

Baker M, Nixon B, Naumovski N, Aitken R . Proteomic insights into the maturation and capacitation of mammalian spermatozoa. Syst Biol Reprod Med. 2012; 58(4):211-7. DOI: 10.3109/19396368.2011.639844. View

11.

Goss D, Vasilescu S, Sacks G, Gardner D, Warkiani M . Microfluidics facilitating the use of small extracellular vesicles in innovative approaches to male infertility. Nat Rev Urol. 2022; 20(2):66-95. DOI: 10.1038/s41585-022-00660-8. View

12.

Khatun A, Rahman M, Pang M . Clinical assessment of the male fertility. Obstet Gynecol Sci. 2018; 61(2):179-191. PMC: 5854897. DOI: 10.5468/ogs.2018.61.2.179. View

13.

Ding Y, Hu Q, Gan J, Zang X, Gu T, Wu Z . Effect of miR-143-3p from Extracellular Vesicles of Porcine Uterine Luminal Fluid on Porcine Trophoblast Cells. Animals (Basel). 2022; 12(23). PMC: 9736583. DOI: 10.3390/ani12233402. View

14.

Bachurski D, Schuldner M, Nguyen P, Malz A, Reiners K, Grenzi P . Extracellular vesicle measurements with nanoparticle tracking analysis - An accuracy and repeatability comparison between NanoSight NS300 and ZetaView. J Extracell Vesicles. 2019; 8(1):1596016. PMC: 6450530. DOI: 10.1080/20013078.2019.1596016. View

15.

James E, Carrell D, Aston K, Jenkins T, Yeste M, Salas-Huetos A . The Role of the Epididymis and the Contribution of Epididymosomes to Mammalian Reproduction. Int J Mol Sci. 2020; 21(15). PMC: 7432785. DOI: 10.3390/ijms21155377. View

16.

Ghafourian M, Mahdavi R, Jonoush Z, Sadeghi M, Ghadiri N, Farzaneh M . The implications of exosomes in pregnancy: emerging as new diagnostic markers and therapeutics targets. Cell Commun Signal. 2022; 20(1):51. PMC: 9004059. DOI: 10.1186/s12964-022-00853-z. View

17.

Alvarez-Rodriguez M, Martinez-Pastor F . Molecular Determinants of Seminal Plasma on Sperm Biology and Fertility. Int J Mol Sci. 2021; 22(7). PMC: 8037708. DOI: 10.3390/ijms22073555. View

18.

Willforss J, Morrell J, Resjo S, Hallap T, Padrik P, Siino V . Stable bull fertility protein markers in seminal plasma. J Proteomics. 2021; 236:104135. DOI: 10.1016/j.jprot.2021.104135. View

19.

Wang L, Zhang J, Xia M, Liu C, Zu X, Zhong J . High Mobility Group A1 (HMGA1): Structure, Biological Function, and Therapeutic Potential. Int J Biol Sci. 2022; 18(11):4414-4431. PMC: 9295051. DOI: 10.7150/ijbs.72952. View

20.

Park Y, Kim J, You Y, Pang M . Proteomic revolution to improve tools for evaluating male fertility in animals. J Proteome Res. 2013; 12(11):4738-47. DOI: 10.1021/pr400639x. View