First sex modification case in equine cloning.
Abstract: Somatic cell nuclear transfer (SCNT) is an asexual reproductive technique where cloned offspring contain the same genetic material as the original donor. Although this technique preserves the sex of the original animal, the birth of sex-reversed offspring has been reported in some species. Here, we report for the first time the birth of a female foal generated by SCNT of a male nuclear donor. After a single SCNT procedure, 16 blastocysts were obtained and transferred to eight recipient mares, resulting in the birth of two clones: one male and one female. Both animals had identical genetic profiles, as observed in the analysis of 15-horse microsatellite marker panel, which confirmed they are indeed clones of the same animal. Cytogenetic analysis and fluorescent in situ hybridization using X and Y specific probes revealed a 63,X chromosome set in the female offspring, suggesting a spontaneous Y chromosome loss. The identity of the lost chromosome in the female was further confirmed through PCR by observing the presence of X-linked markers and absence of Y-linked markers. Moreover, cytogenetic and molecular profiles were analyzed in blood and skin samples to detect a possible mosaicism in the female, but results showed identical chromosomal constitutions. Although the cause of the spontaneous chromosome loss remains unknown, the possibility of equine sex reversal by SCNT holds great potential for the preservation of endangered species, development of novel breeding techniques, and sportive purposes.
Copyright: © 2023 Suvá et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Publication Date: 2023-01-04 PubMed ID: 36598913PubMed Central: PMC9812313DOI: 10.1371/journal.pone.0279869Google Scholar: Lookup
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Summary
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The research highlights the first instance of unintentional sex reversal in a cloned horse, with a female foal being produced from the cloning of a male donor.
Methodology and Results
- The researchers used Somatic cell nuclear transfer (SCNT) for the cloning process. This is a lab technique that creates an ovum with a donor nucleus and it does not involve sexual reproduction.
- After conducting a single SCNT procedure, 16 blastocysts, or clusters of cells developed from the fertilized ovum, were obtained.
- These blastocysts were then transferred to eight recipient mares, yielding two clones: one male and one female.
Genetic Analysis
- The scientists conducted an analysis using a 15-horse microsatellite marker panel on the two clones and found both animals to have identical genetic profiles, verifying that they are true clones of the same donor animal.
- By conducting a cytogenetic analysis and fluorescent in situ hybridization using X and Y specific probes, it was revealed that the female clone had a 63,X chromosome set. This suggests there was a spontaneous loss of the Y chromosome, resulting in the unexpected sex reversal.
- PCR (polymerase chain reaction) testing was further used to confirm the identity of the lost chromosome in the female. The female clone displayed the presence of X-linked genetic markers and absence of Y-linked markers.
Inspection of Potential Mosaicism
- The researchers then undertook cytogenetic and molecular profile analysis on the blood and skin samples of the female clone to check for possible mosaicism, a condition where an individual has cells with different genotypes. However, the results displayed identical chromosomal constitutions, ruling out mosaicism.
Implications of the Study
- The spontaneous sex reversal observed in this study, while unexpected, suggests potential for the manipulation of sex in equine cloning. This could have significant implications in the preservation of endangered species or development of new breeding techniques.
- The cause behind the spontaneous chromosome loss remains undetermined, which necessitates further study to understand the phenomenon better.
Cite This Article
APA
Suvá M, Arnold VH, Wiedenmann EA, Jordan R, Galvagno E, Martínez M, Vichera GD.
(2023).
First sex modification case in equine cloning.
PLoS One, 18(1), e0279869.
https://doi.org/10.1371/journal.pone.0279869 Publication
Researcher Affiliations
- Kheiron Biotech S.A., Pilar, Buenos Aires, Argentina.
- Kheiron Biotech S.A., Pilar, Buenos Aires, Argentina.
- Kheiron Biotech S.A., Pilar, Buenos Aires, Argentina.
- Kheiron Biotech S.A., Pilar, Buenos Aires, Argentina.
- Kheiron Biotech S.A., Pilar, Buenos Aires, Argentina.
- Laboratorio de Genética Aplicada, Sociedad Rural Argentina, Buenos Aires, Argentina.
- Kheiron Biotech S.A., Pilar, Buenos Aires, Argentina.
MeSH Terms
- Male
- Animals
- Horses / genetics
- Female
- In Situ Hybridization, Fluorescence
- Cloning, Organism / veterinary
- Nuclear Transfer Techniques / veterinary
- X Chromosome / genetics
- Cloning, Molecular
Conflict of Interest Statement
The authors have declared that no competing interests exist.
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Citations
This article has been cited 1 times.- Inoue K. Mouse somatic cell nuclear transfer: What has changed and remained unchanged in 25 years.. J Reprod Dev 2023 Jun 6;69(3):129-138.
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