Extracellular Reactive Oxygen Species (ROS) Production in Fresh Donkey Sperm Exposed to Reductive Stress, Oxidative Stress and NETosis.
Abstract: Jenny shows a large endometrial reaction after semen influx to the uterus with a large amount of polymorphonuclear neutrophils (PMN) migrating into the uterine lumen. PMN act as a sperm selection mechanism through phagocytosis and NETosis (DNA extrudes and, together with proteins, trap spermatozoa). While a reduced percentage of spermatozoa are phagocytosed by PMN, most are found to be attached to neutrophil extracellular traps (NETs). This selection process together with sperm metabolism produces a large amount of reactive oxygen species (ROS) that influence the reproductive success. The present study aimed to determine the extracellular ROS production in both sperm and PMN. With this purpose, (1) donkey sperm were exposed to reductive and oxidative stresses, through adding different concentrations of reduced glutathione (GSH) and hydrogen peroxide (HO), respectively; and (2) PMN were subjected to NETosis in the presence of the whole semen, sperm, seminal plasma (SP) or other activators such as formyl-methionyl-leucyl-phenylalanine (FMLP). Extracellular ROS production (measured as HO levels) was determined with the Amplex Red Hydrogen Peroxide/Peroxidase Assay Kit. Donkey sperm showed more resilience to oxidative stress than to the reductive one, and GSH treatments led to greater HO extracellular production. Moreover, not only did SP appear to be the main inducer of NETosis in PMN, but it was also able to maintain the extracellular HO levels produced by sperm and NETosis.
Publication Date: 2021-08-27 PubMed ID: 34572999PubMed Central: PMC8470534DOI: 10.3390/antiox10091367Google Scholar: Lookup
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Summary
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The research article examines how exposure to different types of stress affects the production of reactive oxygen species (ROS) in fresh donkey sperm. The researchers also study the role of different components in the process of netosis, a method by which neutrophils trap and destroy pathogens.
Methodology
- The researchers conducted two main experiments. In the first one, they exposed donkey sperm to reductive stress and oxidative stress. They did this by adding varying amounts of reduced glutathione (GSH), known for its antioxidant properties, and hydrogen peroxide (HO), a reactive molecule involved in oxidative stress.
- In the second experiment, the researchers subjected polymorphonuclear neutrophils (PMN) to netosis. They used whole semen, sperm, seminal plasma (SP) and formyl-methionyl-leucyl-phenylalanine (FMLP), an activator of neutrophils, to stimulate this process.
- The researchers measured extracellular ROS production (measured as HO levels) by using the Amplex Red Hydrogen Peroxide/Peroxidase Assay Kit.
Findings
- The researchers found that donkey sperm showed higher resilience to oxidative stress as compared to reductive stress. They established this by noting more production of HO under the influence of antioxidant GSH treatments versus those involving oxidative stress through HO.
- Additionally, the research team discovered that seminal plasma (SP) appeared to be the primary trigger of netosis in PMN, an important physiological response of the immune system involved in trapping and immobilizing pathogens.
- The results also indicated that seminal plasma played a crucial role in maintaining the levels of extracellular hydrogen peroxide produced by sperm and the process of netosis.
Significance
- This study provides new insights into the production of ROS in both sperm and PMN, which can help in developing strategies to optimize reproductive success.
- The findings also underline the significant roles of antioxidant and oxidative stresses in sperm resilience and PMN’s netosis process, shedding light on their potential implications for male fertility and immune response mechanisms.
Cite This Article
APA
Yánez-Ortiz I, Catalán J, Mateo-Otero Y, Dordas-Perpinyà M, Gacem S, Yeste N, Bassols A, Yeste M, Miró J.
(2021).
Extracellular Reactive Oxygen Species (ROS) Production in Fresh Donkey Sperm Exposed to Reductive Stress, Oxidative Stress and NETosis.
Antioxidants (Basel), 10(9), 1367.
https://doi.org/10.3390/antiox10091367 Publication
Researcher Affiliations
- Equine Reproduction Service, Department of Animal Medicine and Surgery, Faculty of Veterinary Sciences, Autonomous University of Barcelona, E-08193 Bellaterra, Cerdanyola del Vallès, Spain.
- Biotechnology of Animal and Human Reproduction (TechnoSperm), Institute of Food and Agricultural Technology, University of Girona, E-17003 Girona, Spain.
- Unit of Cell Biology, Department of Biology, Faculty of Sciences, University of Girona, E-17003 Girona, Spain.
- Equine Reproduction Service, Department of Animal Medicine and Surgery, Faculty of Veterinary Sciences, Autonomous University of Barcelona, E-08193 Bellaterra, Cerdanyola del Vallès, Spain.
- Biotechnology of Animal and Human Reproduction (TechnoSperm), Institute of Food and Agricultural Technology, University of Girona, E-17003 Girona, Spain.
- Unit of Cell Biology, Department of Biology, Faculty of Sciences, University of Girona, E-17003 Girona, Spain.
- Biotechnology of Animal and Human Reproduction (TechnoSperm), Institute of Food and Agricultural Technology, University of Girona, E-17003 Girona, Spain.
- Unit of Cell Biology, Department of Biology, Faculty of Sciences, University of Girona, E-17003 Girona, Spain.
- Equine Reproduction Service, Department of Animal Medicine and Surgery, Faculty of Veterinary Sciences, Autonomous University of Barcelona, E-08193 Bellaterra, Cerdanyola del Vallès, Spain.
- Equine Reproduction Service, Department of Animal Medicine and Surgery, Faculty of Veterinary Sciences, Autonomous University of Barcelona, E-08193 Bellaterra, Cerdanyola del Vallès, Spain.
- Department of Biochemistry and Molecular Biology, Faculty of Veterinary Sciences, Autonomous University of Barcelona, E-08193 Bellaterra, Cerdanyola del Vallès, Spain.
- Department of Biochemistry and Molecular Biology, Faculty of Veterinary Sciences, Autonomous University of Barcelona, E-08193 Bellaterra, Cerdanyola del Vallès, Spain.
- Biotechnology of Animal and Human Reproduction (TechnoSperm), Institute of Food and Agricultural Technology, University of Girona, E-17003 Girona, Spain.
- Unit of Cell Biology, Department of Biology, Faculty of Sciences, University of Girona, E-17003 Girona, Spain.
- Equine Reproduction Service, Department of Animal Medicine and Surgery, Faculty of Veterinary Sciences, Autonomous University of Barcelona, E-08193 Bellaterra, Cerdanyola del Vallès, Spain.
Conflict of Interest Statement
The authors declare no conflict of interest.
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