Cargando…

Cryoprotective Effects of Ergothioneine and Isoespintanol on Canine Semen

SIMPLE SUMMARY: Cryopreserving dog semen allows the long-term availability of male gametes for future artificial insemination and other assisted reproductive techniques. However, freezing causes irreversible damage to sperm that can affect its ability to fertilize and generate a viable pregnancy. Sp...

Descripción completa

Detalles Bibliográficos
Autores principales: Usuga, Alexandra, Tejera, Irene, Gómez, Jorge, Restrepo, Oliver, Rojano, Benjamín, Restrepo, Giovanni
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8532791/
https://www.ncbi.nlm.nih.gov/pubmed/34679779
http://dx.doi.org/10.3390/ani11102757
_version_ 1784587153799380992
author Usuga, Alexandra
Tejera, Irene
Gómez, Jorge
Restrepo, Oliver
Rojano, Benjamín
Restrepo, Giovanni
author_facet Usuga, Alexandra
Tejera, Irene
Gómez, Jorge
Restrepo, Oliver
Rojano, Benjamín
Restrepo, Giovanni
author_sort Usuga, Alexandra
collection PubMed
description SIMPLE SUMMARY: Cryopreserving dog semen allows the long-term availability of male gametes for future artificial insemination and other assisted reproductive techniques. However, freezing causes irreversible damage to sperm that can affect its ability to fertilize and generate a viable pregnancy. Sperm alterations are partly attributed to oxidation produced by reactive oxygen species (ROS); therefore, antioxidants have been included as extenders for seminal cryopreservation. The unconventional natural antioxidants might reduce deleterious changes in cryopreserved dog sperm; therefore, we evaluated the effects of cryopreservation with the antioxidants ergothioneine and isoespintanol on thawed canine sperm. Various concentrations of both antioxidants improved the movement capacity and structure of thawed spermatozoa, possibly by reducing ROS production. The unconventional antioxidants isoespintanol and ergothioneine improved the quality of cryopreserved canine semen and hence improved assisted canine reproduction. ABSTRACT: Sperm undergo oxidative stress due to excessive production of reactive oxygen species (ROS) during cryopreservation. Some unconventional natural antioxidants can reduce ROS-induced changes in cryopreserved canine sperm. This study aimed to identify the cryoprotective effects of ergothioneine and isoespintanol on the quality of thawed canine semen. Twelve ejaculates from six dogs were cryopreserved in a tris-yolk extender without (control) or with 50 (E50), 100 (E100), or 150 (E150) µM ergothioneine or 20 (I20), 40 (I40), or 60 (I60) µM isoespintanol. We evaluated the motility and kinetics of thawed sperm using computerized analysis; determined morphology by eosin-nigrosin staining; functional membrane integrity using hypoosmotic tests, and structural membrane and acrosome integrity; mitochondrial membrane potential by fluorescence microscopy; and ROS production by spectrophotometry. Data were statistically analyzed using mixed models and Tukey tests. E100 increased total (60.6% vs. 49.6%) and progressive (26.4% vs. 20.1%) motility, straight line velocity (41.3 vs. 35.9 µm/s), and rapid sperm (17.6% vs. 12.3%) compared with controls. However, E150 reduced the numbers of hyperactive sperm. E100, I40, and I60 reduced the abnormal morphology and ROS production, and all concentrations of both antioxidants increased acrosomal integrity. We concluded that ergothioneine and isoespintanol reduce deleterious sperm alterations and oxidative stress in thawed canine semen.
format Online
Article
Text
id pubmed-8532791
institution National Center for Biotechnology Information
language English
publishDate 2021
publisher MDPI
record_format MEDLINE/PubMed
spelling pubmed-85327912021-10-23 Cryoprotective Effects of Ergothioneine and Isoespintanol on Canine Semen Usuga, Alexandra Tejera, Irene Gómez, Jorge Restrepo, Oliver Rojano, Benjamín Restrepo, Giovanni Animals (Basel) Article SIMPLE SUMMARY: Cryopreserving dog semen allows the long-term availability of male gametes for future artificial insemination and other assisted reproductive techniques. However, freezing causes irreversible damage to sperm that can affect its ability to fertilize and generate a viable pregnancy. Sperm alterations are partly attributed to oxidation produced by reactive oxygen species (ROS); therefore, antioxidants have been included as extenders for seminal cryopreservation. The unconventional natural antioxidants might reduce deleterious changes in cryopreserved dog sperm; therefore, we evaluated the effects of cryopreservation with the antioxidants ergothioneine and isoespintanol on thawed canine sperm. Various concentrations of both antioxidants improved the movement capacity and structure of thawed spermatozoa, possibly by reducing ROS production. The unconventional antioxidants isoespintanol and ergothioneine improved the quality of cryopreserved canine semen and hence improved assisted canine reproduction. ABSTRACT: Sperm undergo oxidative stress due to excessive production of reactive oxygen species (ROS) during cryopreservation. Some unconventional natural antioxidants can reduce ROS-induced changes in cryopreserved canine sperm. This study aimed to identify the cryoprotective effects of ergothioneine and isoespintanol on the quality of thawed canine semen. Twelve ejaculates from six dogs were cryopreserved in a tris-yolk extender without (control) or with 50 (E50), 100 (E100), or 150 (E150) µM ergothioneine or 20 (I20), 40 (I40), or 60 (I60) µM isoespintanol. We evaluated the motility and kinetics of thawed sperm using computerized analysis; determined morphology by eosin-nigrosin staining; functional membrane integrity using hypoosmotic tests, and structural membrane and acrosome integrity; mitochondrial membrane potential by fluorescence microscopy; and ROS production by spectrophotometry. Data were statistically analyzed using mixed models and Tukey tests. E100 increased total (60.6% vs. 49.6%) and progressive (26.4% vs. 20.1%) motility, straight line velocity (41.3 vs. 35.9 µm/s), and rapid sperm (17.6% vs. 12.3%) compared with controls. However, E150 reduced the numbers of hyperactive sperm. E100, I40, and I60 reduced the abnormal morphology and ROS production, and all concentrations of both antioxidants increased acrosomal integrity. We concluded that ergothioneine and isoespintanol reduce deleterious sperm alterations and oxidative stress in thawed canine semen. MDPI 2021-09-22 /pmc/articles/PMC8532791/ /pubmed/34679779 http://dx.doi.org/10.3390/ani11102757 Text en © 2021 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Usuga, Alexandra
Tejera, Irene
Gómez, Jorge
Restrepo, Oliver
Rojano, Benjamín
Restrepo, Giovanni
Cryoprotective Effects of Ergothioneine and Isoespintanol on Canine Semen
title Cryoprotective Effects of Ergothioneine and Isoespintanol on Canine Semen
title_full Cryoprotective Effects of Ergothioneine and Isoespintanol on Canine Semen
title_fullStr Cryoprotective Effects of Ergothioneine and Isoespintanol on Canine Semen
title_full_unstemmed Cryoprotective Effects of Ergothioneine and Isoespintanol on Canine Semen
title_short Cryoprotective Effects of Ergothioneine and Isoespintanol on Canine Semen
title_sort cryoprotective effects of ergothioneine and isoespintanol on canine semen
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8532791/
https://www.ncbi.nlm.nih.gov/pubmed/34679779
http://dx.doi.org/10.3390/ani11102757
work_keys_str_mv AT usugaalexandra cryoprotectiveeffectsofergothioneineandisoespintanoloncaninesemen
AT tejerairene cryoprotectiveeffectsofergothioneineandisoespintanoloncaninesemen
AT gomezjorge cryoprotectiveeffectsofergothioneineandisoespintanoloncaninesemen
AT restrepooliver cryoprotectiveeffectsofergothioneineandisoespintanoloncaninesemen
AT rojanobenjamin cryoprotectiveeffectsofergothioneineandisoespintanoloncaninesemen
AT restrepogiovanni cryoprotectiveeffectsofergothioneineandisoespintanoloncaninesemen