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ORIGINAL RESEARCH article

Front. Cell Dev. Biol.
Sec. Molecular and Cellular Reproduction
Volume 12 - 2024 | doi: 10.3389/fcell.2024.1473825
This article is part of the Research Topic Methods for the Study of Sperm Motility and Function View all 6 articles

Differential abundance of microRNAs in seminal plasma extracellular vesicles (EVs) in Sahiwal cattle bull related to male fertility

Provisionally accepted
Vitika Chauhan Vitika Chauhan 1Poonam Kashyap Poonam Kashyap 1Jatinder s. Chera Jatinder s. Chera 1Ankit Pal Ankit Pal 1Aditya Patel Aditya Patel 1Seema Karanwal Seema Karanwal 1Shiva Badrhan Shiva Badrhan 1Fanny Josan Fanny Josan 1Subhash Solanki Subhash Solanki 1Mukesh Bhakat Mukesh Bhakat 2Tirtha K. Datta Tirtha K. Datta 3Rakesh Kumar Rakesh Kumar 1*
  • 1 National Dairy Research Institute (ICAR), Karnal, India
  • 2 Central Institute for Research on Goats (ICAR), Meerut, Uttar Pradesh, India
  • 3 Central Institute for Research on Buffaloes (ICAR), Hisar, Haryana, India

The final, formatted version of the article will be published soon.

    Sahiwal cattle, known for their high milk yield, are propagated through artificial insemination (AI) using male germplasm, largely contingent on semen quality. Spermatozoa, produced in the testes, carry genetic information and molecular signals essential for successful fertilization.Seminal plasma, in addition to sperm, contains nano-sized lipid-bound extracellular vesicles (SP-EVs) that carry key biomolecules, including fertility-related miRNAs, which are essential for bull fertility. The current study focused on miRNA profiling of SP-EVs from high-fertile (HF) and low-fertile (LF) Sahiwal bulls. SP-EVs were isolated using size exclusion chromatography (SEC) and characterized by dynamic light scattering (DLS) and nanoparticle tracking analysis (NTA). Western blotting detected the EV-specific protein markers TSG101 and CD63. The DLS analysis showed SP-EV sizes of 170-180 nm in HF and 130-140 nm in LF samples. The NTA revealed particle concentrations of 5.76 × 10 10 to 5.86 × 10 11 particles/ml in HF and 5.31 × 10 10 to 2.70 × 10 11 particles/ml in LF groups, with no significant differences in size and concentration between HF and LF. High-throughput miRNA sequencing identified 310 miRNAs in SP-EVs from both groups, with 61 upregulated and 119 downregulated in HF bull. Further analysis identified 41 miRNAs with significant fold changes and p-values, including bta-miR-1246, bta-miR-195, bta-miR-339b, and bta-miR-199b, which were analyzed for target gene prediction. Gene Ontology (GO) and KEGG pathway analyses indicated that these miRNAs target genes involved in transcription regulation, ubiquitin-dependent endoplasmic reticulum-associated degradation (ERAD) pathways, and signalling pathways.Functional exploration revealed that these genes play roles in spermatogenesis, motility, acrosome reactions, and inflammatory responses. qPCR analysis showed that bta-miR-195 had 80% higher expression in HF spermatozoa compared to LF, suggesting its association with fertility status (p<0.05). In conclusion, this study elucidates the miRNA cargoes in SP-EVs as indicators of Sahiwal bull fertility, highlighting bta-miR-195 as a potential fertility factor among the various miRNAs identified.

    Keywords: Sahiwal, seminal plasma extracellular vesicles, RNA-Seq, miRNA, Bull fertility

    Received: 31 Jul 2024; Accepted: 18 Sep 2024.

    Copyright: © 2024 Chauhan, Kashyap, Chera, Pal, Patel, Karanwal, Badrhan, Josan, Solanki, Bhakat, Datta and Kumar. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

    * Correspondence: Rakesh Kumar, National Dairy Research Institute (ICAR), Karnal, India

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