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Biology2026; 15(1); 100; doi: 10.3390/biology15010100

Transcriptome Sequencing and Differential Analysis of Testes of 1-Year-Old and 3-Year-Old Kazakh Horses.

Abstract: The Kazakh horse is an outstanding dual-purpose dairy and meat breed in China, characterized by early maturity, tolerance to coarse feed, and strong stress resistance. Previous studies have examined gene expression patterns in the testicular tissues of Kazakh horses at different age stages, but the molecular mechanisms regulating testicular sexual maturation remain unclear. To address this gap, this study conducted HE staining and in-depth transcriptome sequencing analysis of Kazakh horse testicular tissue before and after sexual maturity. HE staining showed that the G3 group had well-formed seminiferous tubule lumens, dense interstitial cells, and visible early spermatocytes and spermatozoa, indicating structural maturation. (G1 group: pre-sexual maturity; G3 group: post-sexual maturity), with four biological replicates per group ( = 4). Differentially expressed genes (DEGs) were called using the criteria of |log(fold change)| ≥ 1.5 and adjusted -value ≤ 0.05. A total of 3054 differentially expressed genes (DEGs), including , , , , and , were identified in the G1 and G3 groups. Among these, 402 genes showed upregulation and 2652 genes showed downregulation. GO annotation and KEGG enrichment analysis of DEGs revealed their predominant enrichment in the following categories: signaling pathways such as Focal adhesion, Pathways in cancer, and the PI3K-Akt signaling pathway. RT-qPCR validation confirmed the accuracy of the transcriptomic sequencing data. This study further elucidates the differentially expressed genes and associated signaling pathways in Kazakh stallion testes tissue before and after sexual maturity, providing a theoretical foundation and data reference for enhancing reproductive efficiency in equids and promoting biological processes such as testes development and spermatogenesis.
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Publication Date: 2026-01-04 PubMed ID: 41514941PubMed Central: PMC12784889DOI: 10.3390/biology15010100Google 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.

Overview

  • This study analyzed the changes in gene expression in the testes of Kazakh horses before and after sexual maturity to better understand the molecular processes regulating testicular development and function.

Background

  • The Kazakh horse is a notable dual-purpose breed used for both dairy and meat in China.
  • It is known for early sexual maturity, the ability to consume coarse feed, and strong resistance to environmental stress.
  • Prior research has studied gene expression in testicular tissues at various ages, but the molecular mechanisms of sexual maturation remained unclear.

Objectives

  • To investigate the differential gene expression in the testes of Kazakh horses at pre-sexual maturity (1-year-old, G1 group) and post-sexual maturity (3-year-old, G3 group).
  • To elucidate the biological pathways and genes involved in testicular sexual maturation and spermatogenesis.
  • To provide insights that could improve reproductive efficiency in horses.

Methods

  • Histological Analysis: Hematoxylin and eosin (HE) staining was performed on testicular tissues from both age groups.
  • Transcriptome Sequencing: RNA sequencing was carried out to analyze gene expression profiles, with four biological replicates per group.
  • Differential Gene Expression Analysis: DEGs were identified using thresholds of |log(fold change)| ≥ 1.5 and adjusted p-value ≤ 0.05.
  • Functional Annotation: Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses were conducted to determine involved biological functions and pathways.
  • Validation: Quantitative real-time PCR (RT-qPCR) was used to validate the sequencing results.

Results

  • Histology:
    • In the 3-year-old group (G3), seminiferous tubules were well-formed with visible lumens.
    • Dense interstitial cells and early spermatocytes/spermatozoa were evident, indicating structural and functional maturity.
    • The 1-year-old group (G1) showed immature testicular architecture.
  • Differential Expression:
    • A total of 3,054 DEGs were identified between the G1 and G3 groups.
    • 402 genes were upregulated in the post-maturity testes, while 2,652 genes were downregulated.
  • Functional Enrichment:
    • GO and KEGG analyses showed enrichment in signaling pathways related to testicular development and function.
    • Key pathways included focal adhesion, cancer pathways, and PI3K-Akt signaling pathway, which are important for cell communication, proliferation, and survival.
  • Validation:
    • RT-qPCR confirmed the expression patterns observed in the transcriptome sequencing, supporting data reliability.

Conclusions and Implications

  • The study identified critical genes and pathways differentially expressed before and after sexual maturity in Kazakh horse testes.
  • These findings enhance understanding of molecular mechanisms underlying testicular maturation and spermatogenesis in horses.
  • The data serve as a valuable foundation for improving reproductive performance in equine species.
  • The elucidated pathways might be targeted to promote testicular development and biological functions related to fertility in breeding programs.

Cite This Article

APA
Liu J, Yang Y, Wen L, Wen M, Zeng Y, Ren W, Yao X. (2026). Transcriptome Sequencing and Differential Analysis of Testes of 1-Year-Old and 3-Year-Old Kazakh Horses. Biology (Basel), 15(1), 100. https://doi.org/10.3390/biology15010100

Publication

Biology
ISSN: 2079-7737
NlmUniqueID: 101587988
Country: Switzerland
Language: English
Volume: 15
Issue: 1
PII: 100

Researcher Affiliations

Liu, Jiahao
  • College of Animal Science, Xinjiang Agricultural University, Urumqi 830052, China.
Yang, Yuting
  • College of Animal Science, Xinjiang Agricultural University, Urumqi 830052, China.
Wen, Liuxiang
  • College of Animal Science, Xinjiang Agricultural University, Urumqi 830052, China.
  • Xinjiang Key Laboratory of Equine Breeding and Exercise Physiology, Urumqi 830052, China.
Wen, Mingyue
  • College of Animal Science, Xinjiang Agricultural University, Urumqi 830052, China.
  • Xinjiang Key Laboratory of Equine Breeding and Exercise Physiology, Urumqi 830052, China.
Zeng, Yaqi
  • College of Animal Science, Xinjiang Agricultural University, Urumqi 830052, China.
  • Xinjiang Key Laboratory of Equine Breeding and Exercise Physiology, Urumqi 830052, China.
Ren, Wanlu
  • College of Animal Science, Xinjiang Agricultural University, Urumqi 830052, China.
  • Xinjiang Key Laboratory of Equine Breeding and Exercise Physiology, Urumqi 830052, China.
Yao, Xinkui
  • College of Animal Science, Xinjiang Agricultural University, Urumqi 830052, China.
  • Xinjiang Key Laboratory of Equine Breeding and Exercise Physiology, Urumqi 830052, China.

Conflict of Interest Statement

All authors declare that they have no conflicts of interest.

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