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Home / Equine Research Bank / Animals (Basel) / Effect of Alternative Splicing Euchromatic Histone Lysine Me...
Animals : an open access journal from MDPI2025; 15(8); doi: 10.3390/ani15081106

Effect of Alternative Splicing Euchromatic Histone Lysine Methyltransferase 2 (EHMT2/G9A) on Spermatogenesis in Mongolian Horses.

Abstract: The epigenetic regulation of gene expression through the covalent modification of histones is crucial for developing germline cells. To study the regulatory role of alternative splicing (AS) of euchromatic histone lysine methyltransferase 2 (EHMT2/G9A) in spermatogenesis in Mongolian horses, this study first examines the localization of the EHMT2 gene in testicular support cells and then predicts the higher-order structures of sequences with and without AS. Two types of lentiviral vectors for overexpression were subsequently constructed for the EHMT2 gene, one with AS and one without, to infect support cells. The proliferation and activity of infected cells were measured using CCK8, and the differential expression of spermatogenesis-related genes in the two types of support cells was analyzed via qRT-PCR. We analyzed the expression of EHMT2 by immunofluorescence staining. EHMT2 was expressed in the nuclei of Sertoli cells. The expression of spermatogenesis-related genes was measured in the two types of cells. The results reveal that the expression levels of the FSH, Stra8, CCNB2, CDC27, NRG1, PPP2R5C, CCNB2, and YWHAZ genes in the AS group were greater than those in the control group. These results indicate that AS events in EHMT2 affect gene expression and thus affect spermatogenesis.
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Publication Date: 2025-04-11 PubMed ID: 40281940PubMed Central: PMC12024092DOI: 10.3390/ani15081106Google Scholar: Lookup
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Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

The study investigates the role of Alternative Splicing (AS) of euchromatic histone lysine methyltransferase 2 (EHMT2/G9A) gene in the process of spermatogenesis (development of sperm) in Mongolian horses. The researchers found that the AS events in EHMT2/G9A significantly influence the expression of spermatogenesis-related genes.

Gene Localization and Higher-Order Structure Prediction

  • The study first determined the location of the EHMT2/G9A gene in the testicular support cells. This gene plays a significant role in epigenetic regulation, which can influence gene expression and help develop germline cells.
  • The researchers used computational techniques to predict the higher-order structures of sequences with AS and those without it. Higher-order structures refer to the three-dimensional shape that protein sequences adopt, which in turn can impact their functionality.

Viral Vector Construction and Cell Infection

  • Experimentally, two types of lentiviral vectors were built for overexpression of the EHMT2/G9A gene—one incorporating the AS and the other without it. Lentiviral vectors are tools commonly used in molecular biology to deliver a gene of interest into cells. Overexpression is a technique used to study the function of a gene by increasing its activity level.
  • These vectors were then used to infect support cells, allowing the researchers to observe the effects on these cells depending on the presence or absence of AS in the EHMT2/G9A gene.

Cell Proliferation Check and Gene Expression Analysis

  • The researchers then measured the proliferation and activity of the infected cells using a CCK8 assay, a commonly used method to evaluate cell metabolic activity and hence, cell viability and proliferation. Cells that are more metabolically active are generally healthier and proliferate more.
  • The expression of various spermatogenesis-related genes in both cells types was then analyzed using quantitative Reverse Transcription Polymerase Chain Reaction (qRT-PCR), a commonly used method for quantifying gene expression levels.
  • The expression of EHMT2/G9A was also verified through immunofluorescence staining.

Outcome and Implications

  • The experiment’s results showed that the expression levels of several genes associated with spermatogenesis were higher in the group of cells with AS. This finding suggests that AS in EHMT2/G9A is critical for the expression of these genes and ultimately, spermatogenesis.
  • This research helps in understanding the molecular mechanisms of spermatogenesis in horses and potentially other mammals, which could be critical for improving reproductive success and managing genetic resources in animal breeding.

Cite This Article

APA
Baatar T, Song D, Weng Y, Wang G, Jin L, Guo R, Li B, Dugarjaviin M. (2025). Effect of Alternative Splicing Euchromatic Histone Lysine Methyltransferase 2 (EHMT2/G9A) on Spermatogenesis in Mongolian Horses. Animals (Basel), 15(8). https://doi.org/10.3390/ani15081106

Publication

Animals : an open access journal from MDPI
ISSN: 2076-2615
NlmUniqueID: 101635614
Country: Switzerland
Language: English
Volume: 15
Issue: 8

Researcher Affiliations

Baatar, Tergel
  • Key Laboratory of Equus Germplasm Innovation (Coconstruction by Ministry and Province), Ministry of Agriculture and Rural Affairs, Hohhot 010018, China.
  • Inner Mongolia Key Laboratory of Equine Science Research and Technology Innovation, Inner Mongolia Agricultural University, Hohhot 010018, China.
  • Equus Research Center, College of Animal Science, Inner Mongolia Agricultural University, Hohhot 010018, China.
Song, Dailing
  • Key Laboratory of Equus Germplasm Innovation (Coconstruction by Ministry and Province), Ministry of Agriculture and Rural Affairs, Hohhot 010018, China.
  • Inner Mongolia Key Laboratory of Equine Science Research and Technology Innovation, Inner Mongolia Agricultural University, Hohhot 010018, China.
  • Equus Research Center, College of Animal Science, Inner Mongolia Agricultural University, Hohhot 010018, China.
Weng, Yajuan
  • Key Laboratory of Equus Germplasm Innovation (Coconstruction by Ministry and Province), Ministry of Agriculture and Rural Affairs, Hohhot 010018, China.
  • Inner Mongolia Key Laboratory of Equine Science Research and Technology Innovation, Inner Mongolia Agricultural University, Hohhot 010018, China.
  • Equus Research Center, College of Animal Science, Inner Mongolia Agricultural University, Hohhot 010018, China.
Wang, Guoqing
  • Key Laboratory of Equus Germplasm Innovation (Coconstruction by Ministry and Province), Ministry of Agriculture and Rural Affairs, Hohhot 010018, China.
  • Inner Mongolia Key Laboratory of Equine Science Research and Technology Innovation, Inner Mongolia Agricultural University, Hohhot 010018, China.
  • Equus Research Center, College of Animal Science, Inner Mongolia Agricultural University, Hohhot 010018, China.
Jin, Liangyi
  • Key Laboratory of Equus Germplasm Innovation (Coconstruction by Ministry and Province), Ministry of Agriculture and Rural Affairs, Hohhot 010018, China.
  • Inner Mongolia Key Laboratory of Equine Science Research and Technology Innovation, Inner Mongolia Agricultural University, Hohhot 010018, China.
  • Equus Research Center, College of Animal Science, Inner Mongolia Agricultural University, Hohhot 010018, China.
Guo, Rui
  • Key Laboratory of Equus Germplasm Innovation (Coconstruction by Ministry and Province), Ministry of Agriculture and Rural Affairs, Hohhot 010018, China.
  • Inner Mongolia Key Laboratory of Equine Science Research and Technology Innovation, Inner Mongolia Agricultural University, Hohhot 010018, China.
  • Equus Research Center, College of Animal Science, Inner Mongolia Agricultural University, Hohhot 010018, China.
Li, Bei
  • Key Laboratory of Equus Germplasm Innovation (Coconstruction by Ministry and Province), Ministry of Agriculture and Rural Affairs, Hohhot 010018, China.
  • Inner Mongolia Key Laboratory of Equine Science Research and Technology Innovation, Inner Mongolia Agricultural University, Hohhot 010018, China.
  • Equus Research Center, College of Animal Science, Inner Mongolia Agricultural University, Hohhot 010018, China.
Dugarjaviin, Manglai
  • Key Laboratory of Equus Germplasm Innovation (Coconstruction by Ministry and Province), Ministry of Agriculture and Rural Affairs, Hohhot 010018, China.
  • Inner Mongolia Key Laboratory of Equine Science Research and Technology Innovation, Inner Mongolia Agricultural University, Hohhot 010018, China.
  • Equus Research Center, College of Animal Science, Inner Mongolia Agricultural University, Hohhot 010018, China.

Grant Funding

  • BR230152 / Program for improving the Scientific Research Ability of Youth Teachers of Inner Mongolia Agricultural University

Conflict of Interest Statement

The authors declare no conflicts of interest.

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