Controlled spermatozoa–oocyte interaction improves embryo quality in sheep

Abstract The current protocols of in vitro fertilization and culture in sheep rely on paradigms established more than 25 years ago, where Metaphase II oocytes are co-incubated with capacitated spermatozoa overnight. While this approach maximizes the number of fertilized oocytes, on the other side it...

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Autores principales:	Debora Agata Anzalone, Luca Palazzese, Marta Czernik, Annalaura Sabatucci, Luca Valbonetti, Emanuele Capra, Pasqualino Loi
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/f8c0b9dd16604d8a99c0b4135c4f7389
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spelling	oai:doaj.org-article:f8c0b9dd16604d8a99c0b4135c4f73892021-11-21T12:17:04ZControlled spermatozoa–oocyte interaction improves embryo quality in sheep10.1038/s41598-021-02000-z2045-2322https://doaj.org/article/f8c0b9dd16604d8a99c0b4135c4f73892021-11-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-02000-zhttps://doaj.org/toc/2045-2322Abstract The current protocols of in vitro fertilization and culture in sheep rely on paradigms established more than 25 years ago, where Metaphase II oocytes are co-incubated with capacitated spermatozoa overnight. While this approach maximizes the number of fertilized oocytes, on the other side it exposes them to high concentration of reactive oxygen species (ROS) generated by active and degenerating spermatozoa, and positively correlates with polyspermy. Here we set up to precisely define the time frame during which spermatozoa effectively penetrates and fertilizes the oocyte, in order to drastically reduce spermatozoa-oocyte interaction. To do that, in vitro matured sheep oocytes co-incubated with spermatozoa in IVF medium were sampled every 30 min (start of incubation time 0) to verify the presence of a fertilizing spermatozoon. Having defined the fertilization time frame (4 h, data from 105 oocytes), we next compared the standard IVF procedures overnight (about 16 h spermatozoa/oocyte exposure, group o/nIVF) with a short one (4 h, group shIVF). A lower polyspermic fertilization (> 2PN) was detected in shIVF (6.5%) compared to o/nIVF (17.8%), P < 0.05. The o/nIVF group resulted in a significantly lower 2-cell stage embryos, than shIVF [34.6% (81/234) vs 50.6% (122/241) respectively, P < 0.001]. Likewise, the development to blastocyst stage confirmed a better quality [29% (70/241) vs 23.5% (55/234), shIVF vs o/nIVF respectively] and an increased Total Cell Number (TCN) in shIVF embryos, compared with o/n ones. The data on ROS have confirmed that its generation is IVF time-dependent, with high levels in the o/nIVF group. Overall, the data suggest that a shorter oocyte-spermatozoa incubation results in an improved embryo production and a better embryo quality, very likely as a consequence of a shorter exposure to the free oxygen radicals and the ensuing oxidative stress imposed by overnight culture.Debora Agata AnzaloneLuca PalazzeseMarta CzernikAnnalaura SabatucciLuca ValbonettiEmanuele CapraPasqualino LoiNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-9 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Debora Agata Anzalone Luca Palazzese Marta Czernik Annalaura Sabatucci Luca Valbonetti Emanuele Capra Pasqualino Loi Controlled spermatozoa–oocyte interaction improves embryo quality in sheep
description	Abstract The current protocols of in vitro fertilization and culture in sheep rely on paradigms established more than 25 years ago, where Metaphase II oocytes are co-incubated with capacitated spermatozoa overnight. While this approach maximizes the number of fertilized oocytes, on the other side it exposes them to high concentration of reactive oxygen species (ROS) generated by active and degenerating spermatozoa, and positively correlates with polyspermy. Here we set up to precisely define the time frame during which spermatozoa effectively penetrates and fertilizes the oocyte, in order to drastically reduce spermatozoa-oocyte interaction. To do that, in vitro matured sheep oocytes co-incubated with spermatozoa in IVF medium were sampled every 30 min (start of incubation time 0) to verify the presence of a fertilizing spermatozoon. Having defined the fertilization time frame (4 h, data from 105 oocytes), we next compared the standard IVF procedures overnight (about 16 h spermatozoa/oocyte exposure, group o/nIVF) with a short one (4 h, group shIVF). A lower polyspermic fertilization (> 2PN) was detected in shIVF (6.5%) compared to o/nIVF (17.8%), P < 0.05. The o/nIVF group resulted in a significantly lower 2-cell stage embryos, than shIVF [34.6% (81/234) vs 50.6% (122/241) respectively, P < 0.001]. Likewise, the development to blastocyst stage confirmed a better quality [29% (70/241) vs 23.5% (55/234), shIVF vs o/nIVF respectively] and an increased Total Cell Number (TCN) in shIVF embryos, compared with o/n ones. The data on ROS have confirmed that its generation is IVF time-dependent, with high levels in the o/nIVF group. Overall, the data suggest that a shorter oocyte-spermatozoa incubation results in an improved embryo production and a better embryo quality, very likely as a consequence of a shorter exposure to the free oxygen radicals and the ensuing oxidative stress imposed by overnight culture.
format	article
author	Debora Agata Anzalone Luca Palazzese Marta Czernik Annalaura Sabatucci Luca Valbonetti Emanuele Capra Pasqualino Loi
author_facet	Debora Agata Anzalone Luca Palazzese Marta Czernik Annalaura Sabatucci Luca Valbonetti Emanuele Capra Pasqualino Loi
author_sort	Debora Agata Anzalone
title	Controlled spermatozoa–oocyte interaction improves embryo quality in sheep
title_short	Controlled spermatozoa–oocyte interaction improves embryo quality in sheep
title_full	Controlled spermatozoa–oocyte interaction improves embryo quality in sheep
title_fullStr	Controlled spermatozoa–oocyte interaction improves embryo quality in sheep
title_full_unstemmed	Controlled spermatozoa–oocyte interaction improves embryo quality in sheep
title_sort	controlled spermatozoa–oocyte interaction improves embryo quality in sheep
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/f8c0b9dd16604d8a99c0b4135c4f7389
work_keys_str_mv	AT deboraagataanzalone controlledspermatozoaoocyteinteractionimprovesembryoqualityinsheep AT lucapalazzese controlledspermatozoaoocyteinteractionimprovesembryoqualityinsheep AT martaczernik controlledspermatozoaoocyteinteractionimprovesembryoqualityinsheep AT annalaurasabatucci controlledspermatozoaoocyteinteractionimprovesembryoqualityinsheep AT lucavalbonetti controlledspermatozoaoocyteinteractionimprovesembryoqualityinsheep AT emanuelecapra controlledspermatozoaoocyteinteractionimprovesembryoqualityinsheep AT pasqualinoloi controlledspermatozoaoocyteinteractionimprovesembryoqualityinsheep
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Controlled spermatozoa–oocyte interaction improves embryo quality in sheep

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