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Animals : an open access journal from MDPI2026; 16(1); 143; doi: 10.3390/ani16010143

Histomorphometric Analysis of the Endometrium of Jennies (Equus asinus) and Mares (Equus caballus) in Estrus: Anatomical Differences and Possible Reproductive Implications.

Abstract: Assisted reproductive techniques are often extrapolated from horses to donkeys, despite poorer fertility outcomes in jennies. This issue has been attributed to unknown uterine species-specific differences. This study compared, through histomorphometry, the endometrium of jennies and mares. Endometrial biopsies (N = 12) were taken from reproductively sound jennies (n = 6) and mares (n = 6) in estrus. Histomorphometric analysis evaluated luminal (LE, µm) and glandular epithelium height (GE, µm), glandular lumen diameter (LD, µm), glandular area (GA, µm), the number of glands (#G), and glandular tissue percentage (GT, %), measured in the stratum compactum (SC) and spongiosum (SS). A total of 30 measurements of glandular size parameters and 10 fields of glandular density parameters per sample were recorded. Results were statistically compared between species (jennies vs. mares), parity status (maiden vs. foaling), and stratum (SC vs. SS). Jennies exhibited higher ( < 0.05) values than mares for LE, LD-SC, GA-SC, and GT-SC. These findings suggest that the histomorphometric features observed in reproductively sound jennies reflect anatomical differences that might partly explain previously observed species differences in post-breeding uterine response. In conclusion, histomorphometry revealed significant endometrial differences between species, with jennies displaying taller luminal epithelium, greater glandular size, and higher glandular tissue percentage in the SC than mares.
Publication Date: 2026-01-04 PubMed ID: 41514829PubMed Central: PMC12784720DOI: 10.3390/ani16010143Google Scholar: Lookup
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  • Journal Article

Summary

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Overview

  • This study compares the microscopic structure of the uterine lining (endometrium) between jennies (female donkeys) and mares (female horses) during estrus (the fertile phase of the reproductive cycle).
  • The researchers aimed to identify anatomical differences that could explain why jennies typically have lower fertility rates than mares when using assisted reproductive techniques.

Introduction and Background

  • Assisted reproductive technologies (ART) such as artificial insemination and embryo transfer are commonly applied to both horses and donkeys.
  • However, fertility outcomes in donkeys (jennies) are often poorer than in horses (mares), leading to the question of whether differences in uterine anatomy might play a role.
  • The endometrium, or uterine lining, is critical for successful reproduction as it supports embryo attachment and development.
  • Histomorphometry is a method used to quantitatively analyze tissue structure by measuring dimensions and areas of specific components under a microscope.

Study Design and Methods

  • Tissue samples were collected via endometrial biopsies from 6 reproductively sound jennies and 6 reproductively sound mares, all in estrus.
  • Histomorphometric parameters measured included:
    • Luminal epithelium height (LE): Thickness of the lining cells facing the uterine lumen.
    • Glandular epithelium height (GE): Thickness of cells lining the uterine glands.
    • Glandular lumen diameter (LD): Size of the glandular cavity.
    • Glandular area (GA): Total size of the glands.
    • Number of glands (#G): Count of glands per sample field.
    • Glandular tissue percentage (GT): Proportion of tissue occupied by glands.
  • Measurements were made separately in two distinct layers of the endometrium:
    • Stratum compactum (SC): The denser, superficial layer.
    • Stratum spongiosum (SS): The deeper, looser connective tissue layer.
  • Multiple measurements were taken per sample to ensure statistical robustness.
  • Data were statistically compared between species (jennies vs. mares), parity status of mares (maiden vs. foaling), and endometrial layers (SC vs. SS).

Key Findings

  • Jennies showed significantly higher values for several parameters compared to mares:
    • Taller luminal epithelium (LE), suggesting a thicker lining facing the uterine cavity.
    • Larger glandular lumen diameter (LD) in the stratum compactum, indicating bigger gland cavities.
    • Greater glandular area (GA) in the SC, meaning overall larger glands.
    • Higher glandular tissue percentage (GT) in the SC, showing a greater proportion of the tissue is comprised of glands.
  • No mention of significant differences in gland numbers (#G) or glandular epithelium height (GE) was reported in the abstract for jennies versus mares.
  • The distinct endometrial architecture in jennies may reflect species-specific adaptations.

Implications and Conclusion

  • The findings provide anatomical evidence that jennies’ uterine linings differ significantly from mares, particularly in having taller luminal epithelium and larger, more abundant glandular tissue in the stratum compactum.
  • These differences could partly explain the poorer fertility outcomes seen in jennies when using reproductive techniques developed based on horse biology.
  • The greater glandular tissue might affect uterine secretions and interactions with embryos post-breeding, potentially influencing uterine response and fertility.
  • Recognizing these species-specific differences highlights the need for tailored reproductive management and protocols in donkeys rather than direct extrapolation from horses.
  • Overall, histomorphometric analysis is a useful method to uncover subtle but important reproductive anatomical distinctions between closely related species.

Cite This Article

APA
Vallejo-Soto P, Dorado J, Herrera-García R, Álvarez-Delgado C, Gómez-Laguna J, Santiago Á, Manrique M, González Ariza A, León Jurado JM, Hidalgo M, Ortiz I. (2026). Histomorphometric Analysis of the Endometrium of Jennies (Equus asinus) and Mares (Equus caballus) in Estrus: Anatomical Differences and Possible Reproductive Implications. Animals (Basel), 16(1), 143. https://doi.org/10.3390/ani16010143

Publication

ISSN: 2076-2615
NlmUniqueID: 101635614
Country: Switzerland
Language: English
Volume: 16
Issue: 1
PII: 143

Researcher Affiliations

Vallejo-Soto, Pilar
  • Veterinary Reproduction Group (AGR-275), Department of Animal Medicine and Surgery, Faculty of Veterinary Medicine, Universidad de Córdoba, Ctra. Madrid-Cadiz, Km 396, 14071 Cordoba, Spain.
Dorado, Jesús
  • Veterinary Reproduction Group (AGR-275), Department of Animal Medicine and Surgery, Faculty of Veterinary Medicine, Universidad de Córdoba, Ctra. Madrid-Cadiz, Km 396, 14071 Cordoba, Spain.
Herrera-García, Rafaela
  • Veterinary Reproduction Group (AGR-275), Department of Animal Medicine and Surgery, Faculty of Veterinary Medicine, Universidad de Córdoba, Ctra. Madrid-Cadiz, Km 396, 14071 Cordoba, Spain.
Álvarez-Delgado, Carmen
  • Department of Anatomy and Comparative Pathology and Toxicology, Pathology and Immunology Group (UCO-PIG), UIC Zoonosis y Enfermedades Emergentes ENZOEM, Universidad de Córdoba, International Excellence Agrifood Campus 'CeiA3', Ctra. Madrid-Cadiz, km 396, 14071 Cordoba, Spain.
Gómez-Laguna, Jaime
  • Department of Anatomy and Comparative Pathology and Toxicology, Pathology and Immunology Group (UCO-PIG), UIC Zoonosis y Enfermedades Emergentes ENZOEM, Universidad de Córdoba, International Excellence Agrifood Campus 'CeiA3', Ctra. Madrid-Cadiz, km 396, 14071 Cordoba, Spain.
Santiago, Álvaro de
  • Military Centre of Equine Breeding of Ecija, C. Nueva, 2, 41400 Ecija, Spain.
Manrique, María
  • Military Centre of Equine Breeding of Ecija, C. Nueva, 2, 41400 Ecija, Spain.
González Ariza, Antonio
  • Agropecuary Provincial Centre, Diputación de Córdoba, Ctra. Madrid-Cadiz, Km 396, 14014 Cordoba, Spain.
León Jurado, José Manuel
  • Agropecuary Provincial Centre, Diputación de Córdoba, Ctra. Madrid-Cadiz, Km 396, 14014 Cordoba, Spain.
Hidalgo, Manuel
  • Veterinary Reproduction Group (AGR-275), Department of Animal Medicine and Surgery, Faculty of Veterinary Medicine, Universidad de Córdoba, Ctra. Madrid-Cadiz, Km 396, 14071 Cordoba, Spain.
Ortiz, Isabel
  • Veterinary Reproduction Group (AGR-275), Department of Animal Medicine and Surgery, Faculty of Veterinary Medicine, Universidad de Córdoba, Ctra. Madrid-Cadiz, Km 396, 14071 Cordoba, Spain.

Grant Funding

  • UCO-13811332-R / FEDER Andalucía 2014-2020
  • PID2020-116090RB-100 / Ministerio de Ciencia e Innovación, Spain
  • FPU23/03472 / the Spanish FPU fellowship (FPU23/03472) from MUNI (Ministerio de Universidades)

Conflict of Interest Statement

The authors declare no conflicts of interest.

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