Exogenous progesterone during in vitro fertilization improves developmental competence of partially cumulus-denuded bovine oocytes

The progesterone (P4) secreted by cumulus cells has gained attention for its role as a possible physiological inducer of sperm acrosome exocytosis. In mammals, it is generally accepted that fertilization rates of oocytes without cumulus are markedly low. This study assessed the integrity of capacita...

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Main Authors: Suqueli García, María Florencia, Gabbanelli, Nadia, Rios, Glenda Laura
Format: info:ar-repo/semantics/artículo
Language:Inglés
Published: Elsevier 2023
Subjects:
Online Access:http://hdl.handle.net/20.500.12123/16015
https://www.sciencedirect.com/science/article/pii/S0093691X2300287X
https://doi.org/10.1016/j.theriogenology.2023.07.036
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author Suqueli García, María Florencia
Gabbanelli, Nadia
Rios, Glenda Laura
author_browse Gabbanelli, Nadia
Rios, Glenda Laura
Suqueli García, María Florencia
author_facet Suqueli García, María Florencia
Gabbanelli, Nadia
Rios, Glenda Laura
author_sort Suqueli García, María Florencia
collection INTA Digital
description The progesterone (P4) secreted by cumulus cells has gained attention for its role as a possible physiological inducer of sperm acrosome exocytosis. In mammals, it is generally accepted that fertilization rates of oocytes without cumulus are markedly low. This study assessed the integrity of capacitated bovine sperm acrosome when exposed to increasing concentrations of P4, and evaluated whether exogenous P4 during in vitro fertilization (IVF) increases the developmental competence of partially cumulus-denuded oocytes in serum-free conditions. After a 4-h capacitation induction, sperm were incubated with increasing concentrations of P4 (0, 0.1, 10 and 100 μM) and evaluated for viability, caspase activation and acrosome status at three different times (4, 5, and 22 h), including the capacitation induction period. Progesterone induced sperm acrosomal exocytosis without compromising sperm viability or activating sperm caspases. Sperm undergoing acrosome reaction exhibited three differential Concanavalin A patterns, corresponding to early, intermediate and late acrosomal exocytosis. The percentage of these patterns significantly increased over time, regardless of P4 concentration, except for those spermatozoa with late acrosomal exocytosis, which only showed an increase at 22 h of incubation. After incubation for 1 h with 100 μM P4, spermatozoa showing intermediate acrosomal exocytosis significantly increased. At 22 h of incubation, the pattern corresponding to early acrosomal exocytosis evidenced a dose-dependent increase. However, prematurely high levels of acrosome reaction induced by 100 μM P4 led to inefficient IVF outcomes (P < 0.05). Therefore, IVF trials with partially cumulus-denuded oocytes were carried out with lower P4 concentrations (0, 0.1, 1, 5, 10 μM). Cleavage rate significantly increased at 1 μM P4, which translated to increased total embryo production after 7 days of in vitro culture (P < 0.05). Significantly higher percentages of expanded blastocysts were observed at both 1 μM and 10 μM P4 as compared to the other experimental conditions. In conclusion, the different patterns of acrosomal exocytosis identified over time by incubation of live sperm with a fluorescent lectin revealed the existence of sperm subpopulations heterogeneous in their physiological states. Moreover, exogenous P4 at 1 μM during IVF improved the developmental competence of partially cumulus-denuded oocytes in serum-free conditions.
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institution Instituto Nacional de Tecnología Agropecuaria (INTA -Argentina)
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publishDate 2023
publishDateRange 2023
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spelling INTA160152023-11-27T11:02:58Z Exogenous progesterone during in vitro fertilization improves developmental competence of partially cumulus-denuded bovine oocytes Suqueli García, María Florencia Gabbanelli, Nadia Rios, Glenda Laura Hormona Esteroide Ovocitos Ganado Espermatozoo In Vitro Progesterona Steroid Hormones Oocytes Livestock Spermatozoa Progesterone The progesterone (P4) secreted by cumulus cells has gained attention for its role as a possible physiological inducer of sperm acrosome exocytosis. In mammals, it is generally accepted that fertilization rates of oocytes without cumulus are markedly low. This study assessed the integrity of capacitated bovine sperm acrosome when exposed to increasing concentrations of P4, and evaluated whether exogenous P4 during in vitro fertilization (IVF) increases the developmental competence of partially cumulus-denuded oocytes in serum-free conditions. After a 4-h capacitation induction, sperm were incubated with increasing concentrations of P4 (0, 0.1, 10 and 100 μM) and evaluated for viability, caspase activation and acrosome status at three different times (4, 5, and 22 h), including the capacitation induction period. Progesterone induced sperm acrosomal exocytosis without compromising sperm viability or activating sperm caspases. Sperm undergoing acrosome reaction exhibited three differential Concanavalin A patterns, corresponding to early, intermediate and late acrosomal exocytosis. The percentage of these patterns significantly increased over time, regardless of P4 concentration, except for those spermatozoa with late acrosomal exocytosis, which only showed an increase at 22 h of incubation. After incubation for 1 h with 100 μM P4, spermatozoa showing intermediate acrosomal exocytosis significantly increased. At 22 h of incubation, the pattern corresponding to early acrosomal exocytosis evidenced a dose-dependent increase. However, prematurely high levels of acrosome reaction induced by 100 μM P4 led to inefficient IVF outcomes (P < 0.05). Therefore, IVF trials with partially cumulus-denuded oocytes were carried out with lower P4 concentrations (0, 0.1, 1, 5, 10 μM). Cleavage rate significantly increased at 1 μM P4, which translated to increased total embryo production after 7 days of in vitro culture (P < 0.05). Significantly higher percentages of expanded blastocysts were observed at both 1 μM and 10 μM P4 as compared to the other experimental conditions. In conclusion, the different patterns of acrosomal exocytosis identified over time by incubation of live sperm with a fluorescent lectin revealed the existence of sperm subpopulations heterogeneous in their physiological states. Moreover, exogenous P4 at 1 μM during IVF improved the developmental competence of partially cumulus-denuded oocytes in serum-free conditions. EEA Balcarce Fil: Suqueli García, María Florencia. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Balcarce; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Instituto de Innovación para la Producción Agropecuaria y el Desarrollo Sostenible; Argentina. Fil: Gabbanelli, Nadia. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Balcarce; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Instituto de Innovación para la Producción Agropecuaria y el Desarrollo Sostenible; Argentina. Fil: Ríos, Glenda Laura. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Balcarce; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Instituto de Innovación para la Producción Agropecuaria y el Desarrollo Sostenible; Argentina. Fil: Buschiazzo, Jorgelina. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Balcarce; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Instituto de Innovación para la Producción Agropecuaria y el Desarrollo Sostenible; Argentina. 2023-11-27T10:58:02Z 2023-11-27T10:58:02Z 2023-11 info:ar-repo/semantics/artículo info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion http://hdl.handle.net/20.500.12123/16015 https://www.sciencedirect.com/science/article/pii/S0093691X2300287X 0093-691X (Print) 1879-3231 (Online) https://doi.org/10.1016/j.theriogenology.2023.07.036 eng info:eu-repograntAgreement/INTA/2019-PD-E5-I107-001, Biotecnologías de la reproducción: mejora de la eficiencia reproductiva en especies de interés zootécnico info:eu-repo/semantics/restrictedAccess http://creativecommons.org/licenses/by-nc-sa/4.0/ Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International (CC BY-NC-SA 4.0) application/pdf Elsevier Theriogenology 211 : 11-18 (November 2023)
spellingShingle Hormona Esteroide
Ovocitos
Ganado
Espermatozoo
In Vitro
Progesterona
Steroid Hormones
Oocytes
Livestock
Spermatozoa
Progesterone
Suqueli García, María Florencia
Gabbanelli, Nadia
Rios, Glenda Laura
Exogenous progesterone during in vitro fertilization improves developmental competence of partially cumulus-denuded bovine oocytes
title Exogenous progesterone during in vitro fertilization improves developmental competence of partially cumulus-denuded bovine oocytes
title_full Exogenous progesterone during in vitro fertilization improves developmental competence of partially cumulus-denuded bovine oocytes
title_fullStr Exogenous progesterone during in vitro fertilization improves developmental competence of partially cumulus-denuded bovine oocytes
title_full_unstemmed Exogenous progesterone during in vitro fertilization improves developmental competence of partially cumulus-denuded bovine oocytes
title_short Exogenous progesterone during in vitro fertilization improves developmental competence of partially cumulus-denuded bovine oocytes
title_sort exogenous progesterone during in vitro fertilization improves developmental competence of partially cumulus denuded bovine oocytes
topic Hormona Esteroide
Ovocitos
Ganado
Espermatozoo
In Vitro
Progesterona
Steroid Hormones
Oocytes
Livestock
Spermatozoa
Progesterone
url http://hdl.handle.net/20.500.12123/16015
https://www.sciencedirect.com/science/article/pii/S0093691X2300287X
https://doi.org/10.1016/j.theriogenology.2023.07.036
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