Horse ooplasm supports in vitro preimplantation development of zebra ICSI and SCNT embryos without compromising YAP1 and SOX2 expression pattern.
Abstract: Several equids have gone extinct and many extant equids are currently considered vulnerable to critically endangered. This work aimed to evaluate whether domestic horse oocytes support preimplantation development of zebra embryos obtained by intracytoplasmic sperm injection (ICSI, zebroid) and cloning, and to study the Hippo signaling pathway during the lineage specification of trophectoderm cells and inner cell mass cells. We first showed that zebra and horse sperm cells induce porcine oocyte activation and recruit maternal SMARCA4 during pronuclear formation. SMARCA4 recruitment showed to be independent of the genetic background of the injected sperm. No differences were found in blastocyst rate of ICSI hybrid (zebra spermatozoon into horse egg) embryos relative to the homospecific horse control group. Interestingly, zebra cloned blastocyst rate was significantly higher at day 8. Moreover, most ICSI and cloned horse and zebra blastocysts showed a similar expression pattern of SOX2 and nuclear YAP1 with the majority of the nuclei positive for YAP1, and most SOX2+ nuclei negative for YAP1. Here we demonstrated that horse oocytes support zebra preimplantation development of both, ICSI and cloned embryos, without compromising development to blastocyst, blastocyst cell number neither the expression of SOX2 and YAP1. Our results support the use of domestic horse oocytes as a model to study in vitro zebra embryos on behalf of preservation of valuable genetic.
Publication Date: 2020-09-11 PubMed ID: 32915925PubMed Central: PMC7485800DOI: 10.1371/journal.pone.0238948Google Scholar: Lookup
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- Journal Article
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Summary
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This research investigates if the oocytes (undeveloped eggs) of a domestic horse can be used to support the growth of zebra embryos that were created through cloning and sperm injection. The study concludes that horse eggs can support the development of zebra embryos, without compromising their eventual development into blastocysts, or affecting the expression of key genes SOX2 and YAP1.
Objective of the Research
- The main objective of this research is to ascertain if horse eggs can play a role in the formation and growth of zebra embryos—either through sperm injection or cloning techniques. This testing is aimed at contributing to the preservation of valuable genetic data for equids (the horse family), especially the species endangered or extinct.
Sperm Injection and Cloning
- Intracytoplasmic sperm injection (ICSI) is a method where a single sperm is injected into an egg. In this study, zebra sperm was injected into horse eggs. The research found that there was no significant difference in the blastocyst (early-stage embryo) rate of these hybrid embryos when compared to those from horse sperm.
- The study also looked at somatic cell nuclear transfer (SCNT), a cloning method, where a zebra cell is introduced into a horse egg. Remarkably, these zebra cloned embryos had a higher blastocyst rate.
Hippo Signaling Pathway
- The researchers examined the Hippo signalling pathway, a system in cells responsible for regulating organ size and cell proliferation, during the initial separation of cells into two groups—trophectoderm cells, which will form the placenta, and cells of the inner cell mass that develop into the fetus.
- The team analysed the expression pattern of two crucial genes— SOX2 and YAP1. SOX2 is important for maintaining embryonic stem cell properties, and YAP1 is crucial in controlling organ size and cell proliferation. No significant differences in expression pattern were detected.
Findings and Implications
- This research demonstrates that horse oocytes can support zebra embryo development from both sperm injection and cloning rituals without negatively affecting their progression into a blastocyst or altering the expression pattern of SOX2 and YAP1 genes.
- These findings suggest that horse eggs could be used as a model to study in-vitro zebra embryo development. The application of this knowledge indicates promising prospects for the preservation and continuation of valuable genetic resources from endangered or extinct equid species.
Cite This Article
APA
Gambini A, Duque Rodríguez M, Rodríguez MB, Briski O, Flores Bragulat AP, Demergassi N, Losinno L, Salamone DF.
(2020).
Horse ooplasm supports in vitro preimplantation development of zebra ICSI and SCNT embryos without compromising YAP1 and SOX2 expression pattern.
PLoS One, 15(9), e0238948.
https://doi.org/10.1371/journal.pone.0238948 Publication
Researcher Affiliations
- Facultad de Agronomía, Cátedra de Producción Equina, Universidad de Buenos Aires, Buenos Aires, Argentina.
- Consejo Nacional de Investigaciones Científicas y Técnicas, Buenos Aires, Argentina.
- Consejo Nacional de Investigaciones Científicas y Técnicas, Buenos Aires, Argentina.
- Facultad de Agronomía, Cátedra de Fisiología Animal, Universidad de Buenos Aires, Buenos Aires, Argentina.
- Consejo Nacional de Investigaciones Científicas y Técnicas, Buenos Aires, Argentina.
- Facultad de Agronomía, Cátedra de Fisiología Animal, Universidad de Buenos Aires, Buenos Aires, Argentina.
- Consejo Nacional de Investigaciones Científicas y Técnicas, Buenos Aires, Argentina.
- Facultad de Agronomía, Cátedra de Fisiología Animal, Universidad de Buenos Aires, Buenos Aires, Argentina.
- Consejo Nacional de Investigaciones Científicas y Técnicas, Buenos Aires, Argentina.
- Facultad de Agronomía y Veterinaria, Cátedra de Producción Equina, Universidad Nacional de Río Cuarto, Río IV, Córdoba, Argentina.
- Fundación Temaikèn, Belén de Escobar, Buenos Aires, Argentina.
- Facultad de Agronomía y Veterinaria, Cátedra de Producción Equina, Universidad Nacional de Río Cuarto, Río IV, Córdoba, Argentina.
- Consejo Nacional de Investigaciones Científicas y Técnicas, Buenos Aires, Argentina.
- Facultad de Agronomía, Cátedra de Fisiología Animal, Universidad de Buenos Aires, Buenos Aires, Argentina.
MeSH Terms
- Adaptor Proteins, Signal Transducing / genetics
- Animals
- Cell Nucleus / physiology
- Cloning, Organism / veterinary
- Cytoplasm / physiology
- Embryo Culture Techniques / veterinary
- Embryonic Development / genetics
- Embryonic Development / physiology
- Endangered Species
- Equidae / embryology
- Equidae / genetics
- Equidae / metabolism
- Female
- Gene Expression Profiling
- Horses / genetics
- Horses / physiology
- In Vitro Techniques
- Male
- Nuclear Transfer Techniques / veterinary
- Oocytes / physiology
- SOXB1 Transcription Factors / genetics
- Sperm Injections, Intracytoplasmic / veterinary
- Sus scrofa
Conflict of Interest Statement
The authors have declared that no competing interests exist.
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