Elamipretide enhances post-thaw rooster sperm quality by mitigating oxidative stress and optimizing mitochondrial function during cryopreservation

Abstract Cryopreservation of rooster sperm is a vital technique in avian reproductive management; however, it is often hindered by oxidative stress induced by reactive oxygen species (ROS) that negatively impact sperm quality during the freezing-thawing process. The present study aimed to investigat...

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Main Authors: Abouzar Najafi, Seyed Davood Sharifi, Ramin Farhadi, Shorsh Hussein Bapir
Format: Article
Language:English
Published: Nature Portfolio 2025-07-01
Series:Scientific Reports
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Online Access:https://doi.org/10.1038/s41598-025-09943-7
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author Abouzar Najafi
Seyed Davood Sharifi
Ramin Farhadi
Shorsh Hussein Bapir
author_facet Abouzar Najafi
Seyed Davood Sharifi
Ramin Farhadi
Shorsh Hussein Bapir
author_sort Abouzar Najafi
collection DOAJ
description Abstract Cryopreservation of rooster sperm is a vital technique in avian reproductive management; however, it is often hindered by oxidative stress induced by reactive oxygen species (ROS) that negatively impact sperm quality during the freezing-thawing process. The present study aimed to investigate the impact of elamipretide, a mitochondria-targeted peptide, on sperm quality post-thaw. Sperm samples from 32-week-old broiler breeder roosters were cryopreserved using a Lake extender buffer with glycerol as the cryoprotectant. Four different concentrations of elamipretide (0, 6, 9, and 12 µmol/L) were tested in combination with the extender. Post-thaw sperm quality was evaluated by assessing motility, kinematic parameters, membrane integrity, mitochondrial activity, ROS levels as a direct marker of oxidative stress, lipid peroxidation, cell viability, and antioxidant enzyme activities (SOD, GPx and TAC). Sperm motility increased significantly at the 6 µmol/L (60.33 ± 1.54) and 9 µmol/L (64.96 ± 1.96) concentrations compared to the control, with the highest straight-line velocity observed at 9 µmol/L (21.21 ± 0.59). Membrane integrity also improved significantly at 9 µmol/L (61.78 ± 2.70) compared to lower doses (36.30 ± 1.64) and decreased at 12 µmol/L (49.57 ± 1.63). ROS production was significantly lower at 6 µmol/L (2.88 ± 0.07). Mitochondrial activity peaked at 9 µmol/L (60.21 ± 1.92), reflecting enhanced cell vitality and function. However, the effects were diminished at 12 µmol/L, indicating toxicity at higher concentrations. This study demonstrates the potential of elamipretide to improve rooster sperm cryopreservation outcomes by mitigating oxidative damage and preserving sperm quality post-thaw.
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spelling doaj-art-6d0b205dbd1d4e5ca70874bf31d7568f2025-08-20T03:03:27ZengNature PortfolioScientific Reports2045-23222025-07-011511910.1038/s41598-025-09943-7Elamipretide enhances post-thaw rooster sperm quality by mitigating oxidative stress and optimizing mitochondrial function during cryopreservationAbouzar Najafi0Seyed Davood Sharifi1Ramin Farhadi2Shorsh Hussein Bapir3Department of Animal and Poultry Science, Faculty of Agricultural Technology, College of Agriculture and Natural Resources, University of TehranDepartment of Animal and Poultry Science, Faculty of Agricultural Technology, College of Agriculture and Natural Resources, University of TehranDepartment of Animal Science, Faculty of Agriculture, University of KurdistanDepartment of Pharmacy, Raparin Technical and Vocational InstituteAbstract Cryopreservation of rooster sperm is a vital technique in avian reproductive management; however, it is often hindered by oxidative stress induced by reactive oxygen species (ROS) that negatively impact sperm quality during the freezing-thawing process. The present study aimed to investigate the impact of elamipretide, a mitochondria-targeted peptide, on sperm quality post-thaw. Sperm samples from 32-week-old broiler breeder roosters were cryopreserved using a Lake extender buffer with glycerol as the cryoprotectant. Four different concentrations of elamipretide (0, 6, 9, and 12 µmol/L) were tested in combination with the extender. Post-thaw sperm quality was evaluated by assessing motility, kinematic parameters, membrane integrity, mitochondrial activity, ROS levels as a direct marker of oxidative stress, lipid peroxidation, cell viability, and antioxidant enzyme activities (SOD, GPx and TAC). Sperm motility increased significantly at the 6 µmol/L (60.33 ± 1.54) and 9 µmol/L (64.96 ± 1.96) concentrations compared to the control, with the highest straight-line velocity observed at 9 µmol/L (21.21 ± 0.59). Membrane integrity also improved significantly at 9 µmol/L (61.78 ± 2.70) compared to lower doses (36.30 ± 1.64) and decreased at 12 µmol/L (49.57 ± 1.63). ROS production was significantly lower at 6 µmol/L (2.88 ± 0.07). Mitochondrial activity peaked at 9 µmol/L (60.21 ± 1.92), reflecting enhanced cell vitality and function. However, the effects were diminished at 12 µmol/L, indicating toxicity at higher concentrations. This study demonstrates the potential of elamipretide to improve rooster sperm cryopreservation outcomes by mitigating oxidative damage and preserving sperm quality post-thaw.https://doi.org/10.1038/s41598-025-09943-7CryopreservationSpermRoosterMitochondria-targetedElamipretide
spellingShingle Abouzar Najafi
Seyed Davood Sharifi
Ramin Farhadi
Shorsh Hussein Bapir
Elamipretide enhances post-thaw rooster sperm quality by mitigating oxidative stress and optimizing mitochondrial function during cryopreservation
Scientific Reports
Cryopreservation
Sperm
Rooster
Mitochondria-targeted
Elamipretide
title Elamipretide enhances post-thaw rooster sperm quality by mitigating oxidative stress and optimizing mitochondrial function during cryopreservation
title_full Elamipretide enhances post-thaw rooster sperm quality by mitigating oxidative stress and optimizing mitochondrial function during cryopreservation
title_fullStr Elamipretide enhances post-thaw rooster sperm quality by mitigating oxidative stress and optimizing mitochondrial function during cryopreservation
title_full_unstemmed Elamipretide enhances post-thaw rooster sperm quality by mitigating oxidative stress and optimizing mitochondrial function during cryopreservation
title_short Elamipretide enhances post-thaw rooster sperm quality by mitigating oxidative stress and optimizing mitochondrial function during cryopreservation
title_sort elamipretide enhances post thaw rooster sperm quality by mitigating oxidative stress and optimizing mitochondrial function during cryopreservation
topic Cryopreservation
Sperm
Rooster
Mitochondria-targeted
Elamipretide
url https://doi.org/10.1038/s41598-025-09943-7
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