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Effect of ethanol and sperm head reducing agents on the preimplantation development of bovine embryos generated by Piezo-ICSI

Published online by Cambridge University Press:  09 December 2025

Víctor Gallardo ,
Luis Aguila ,
María Elena Arias  and
Ricardo Felmer Open the ORCID record for Ricardo Felmer [Opens in a new window]
Víctor Gallardo
Affiliation:
Laboratory of Reproduction, Center of Excellence in Reproductive Biotechnology (CEBIOR-BIOREN), Faculty of Agricultural and Environmental Sciences, Universidad de la Frontera, Temuco, Chile Master of Science Program specializing in Biology of Reproduction, Universidad de La Frontera, Temuco, Chile
Luis Aguila
Affiliation:
Laboratory of Reproduction, Center of Excellence in Reproductive Biotechnology (CEBIOR-BIOREN), Faculty of Agricultural and Environmental Sciences, Universidad de la Frontera, Temuco, Chile
María Elena Arias
Affiliation:
Laboratory of Reproduction, Center of Excellence in Reproductive Biotechnology (CEBIOR-BIOREN), Faculty of Agricultural and Environmental Sciences, Universidad de la Frontera, Temuco, Chile Department of Agricultural Production, Faculty of Agricultural and Environmental Sciences, Universidad de La Frontera, Temuco, Chile
Ricardo Felmer*
Affiliation:
Laboratory of Reproduction, Center of Excellence in Reproductive Biotechnology (CEBIOR-BIOREN), Faculty of Agricultural and Environmental Sciences, Universidad de la Frontera, Temuco, Chile Department of Agricultural Sciences and Natural Resources, Faculty of Agricultural and Environmental Sciences, Universidad de La Frontera, Temuco, Chile
*
Corresponding author: Ricardo Felmer; Email: ricardo.felmer@ufrontera.cl
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Summary

Intracytoplasmic sperm injection (ICSI) is a widely used assisted reproduction technique, but in cattle it faces major challenges due to inefficient oocyte activation after sperm microinjection. This study investigated different oocyte activation strategies and assessed the potential role of reducing agents glutathione (GSH), cysteamine (Cys) and dithiobutylamine (DTBA) to improve sperm head decondensation and embryo development following Piezo-ICSI. Haploid parthenogenetic activation using different ethanol concentrations (1%, 3%, 7% and 10%) failed to yield blastocysts, while diploid activation with ethanol or ionomycin combined with inhibitors significantly improved cleavage (43–55%) and blastocyst rates (14–27%), respectively. However, applying two ethanol pulses was detrimental, reducing both cleavage and blastocysts likely due to toxic overexposure. Sperm head decondensation compounds in Piezo-ICSI showed a high percentage of inactivated oocytes (75% GSH, 55% Cys and 40% DTBA). The highest male pronuclear formation rates were observed in the control without sperm head decondensation (21%) and with DTBA treatment (10%). Despite this, the treatment with Cys resulted in higher developmental potential to the blastocyst stage (22%) comparable to the control (24%). These data suggest that the inclusion of sperm head decondensing agents could represent a promising new strategy for enhancing the early in vitro development of ICSI-generated embryos. However, for this purpose, careful optimization of the concentration and incubation time of these decondensing compounds is essential.

Keywords

Bovineethanoldecondensed compoundsoocyte activationpiezo-ICSI

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Type
Research Article
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Zygote , First View , pp. 1 - 8
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© The Author(s), 2025. Published by Cambridge University Press

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