Apoptotic markers can be used to forecast the freezeability of stallion spermatozoa.
Abstract: In an attempt to identify valuable markers for potential freezeability of the equine spermatozoa, three ejaculates were collected from five Andalusian stallions and frozen using a standard protocol. Before freezing, three apoptotic cell markers were studied by flow cytometry (early changes in sperm membranes, mitochondrial membrane potential and caspase activity). Post-thaw, spermatozoa were again evaluated for these parameters. Sperm kinematics using CASA were also studied before and after freezing and thawing. Receiving operating system curves were used to evaluate the relative value of the apoptotic markers herein studied, as forecast for potential freezeability. From all parameters studied, the outcome of JC-1 (as proportion of spermatozoa showing simultaneously orange and green fluorescence) had the highest diagnostic power. For potentially bad freezers (less than 25% of intact spermatozoa post-thaw), the significant area under the ROC-curve was 0.985, with a 100% sensitivity and 99.8% specificity for a cut off value of 55.7.
Publication Date: 2008-10-17 PubMed ID: 19019584DOI: 10.1016/j.anireprosci.2008.10.005Google Scholar: Lookup
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- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
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This research investigates the use of apoptotic markers to predict the success rate of freezing stallion spermatoa. The study found that a specific apoptotic marker known as JC-1 has the highest diagnostic power in identifying successful freezing of spermatozoa.
Research Process
- The study began by collecting three ejaculates from five Andalusian stallions. These samples were frozen following a standardized protocol.
- Before the freezing process, the researchers examined three apoptotic cell markers (early changes in sperm membranes, mitochondrial membrane potential, and caspase activity).
- After freezing and thawing, the spermatozoa were evaluated again for these three apoptotic markers.
Kinematics and Receiving Operating System Curves
- In addition to the apoptotic marker assessments, the researchers also studied sperm kinematics— that is, the movement characteristics of the sperm—using a Computer Assisted Sperm Analyzer (CASA) both pre-freezing and post-thawing.
- The team then used receiving operating system (ROC) curves to ascertain the relative significance of the studied apoptotic markers as potential indicators for successful freezing.
Results of the Study
- The results showed that, from all studied parameters (including the movement characteristics and the three apoptotic marker results), JC-1 yielded the highest diagnostic power.
- The diagnostic power of JC-1 was determined by the proportion of sperm cells displaying both orange and green fluorescence. The area under the ROC curve was significantly large, at 0.985.
- The study also identified a ‘cut off’ value of 55.7, which demonstrated a sensitivity rate and specificity rate of 100% and 99.8% respectively when used to predict ‘bad freezers’. This term refers to stallions with less than 25% of their sperm showing intact post-thaw.
Implications of the Research
- This research holds potential implications for the field of equine reproduction, as the identified apoptotic marker JC-1 may serve as a reliable diagnostic tool for predicting the success of freezing stallion sperm.
- This could have practical benefits in situations where preservation of stallion sperm is required, such as artificial insemination or conservation efforts for endangered equine species.
Cite This Article
APA
Ortega-Ferrusola C, García BM, Gallardo-Bolaños JM, González-Fernández L, Rodríguez-Martinez H, Tapia JA, Peña FJ.
(2008).
Apoptotic markers can be used to forecast the freezeability of stallion spermatozoa.
Anim Reprod Sci, 114(4), 393-403.
https://doi.org/10.1016/j.anireprosci.2008.10.005 Publication
Researcher Affiliations
- Veterinary Teaching Hospital, Laboratory of Spermatology University of Extremadura, Avd de la Universidad s/n, Cáceres, Spain.
MeSH Terms
- Animals
- Apoptosis
- Caspases / metabolism
- Cell Membrane / ultrastructure
- Cryopreservation / methods
- Cryopreservation / veterinary
- Flow Cytometry
- Horses
- Hot Temperature
- Male
- Membrane Potential, Mitochondrial
- ROC Curve
- Semen Preservation / methods
- Semen Preservation / veterinary
- Sensitivity and Specificity
- Spermatozoa / enzymology
- Spermatozoa / physiology
- Spermatozoa / ultrastructure
Citations
This article has been cited 17 times.- Strassner FM, Demattio L, Siuda M, Malama E, Muffels G, Bollwein H. Relationships Between Metabolism of Cryopreserved Equine Sperm Determined by the Seahorse Analyzer and Sperm Characteristics Measured by Flow Cytometry and Computer-Assisted Analysis of Motility. Vet Sci 2025 Nov 21;12(12).
- Afsar M, Soleimanzadeh A, Khaki A, Ayen E. Improvement of Post-Thaw Quality and In Vivo Fertility of Simmental Bull Spermatozoa Using Ferulic Acid. Vet Med Sci 2024 Nov;10(6):e70064.
- Suo J, Wang J, Zheng Y, Xiao F, Li R, Huang F, Niu P, Zhu W, Du X, He J, Gao Q, Khan A. Recent advances in cryotolerance biomarkers for semen preservation in frozen form-A systematic review. PLoS One 2024;19(5):e0303567.
- Neila-Montero M, Riesco MF, Alvarez M, Montes-Garrido R, Boixo JC, de Paz P, Anel-Lopez L, Anel L. Centrifugal force assessment in ram sperm: identifying species-specific impact. Acta Vet Scand 2021 Nov 4;63(1):42.
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- Gaitskell-Phillips G, Martín-Cano FE, Ortiz-Rodríguez JM, Silva-Rodríguez A, Gil MC, Ortega-Ferrusola C, Peña FJ. In Stallion Spermatozoa, Superoxide Dismutase (Cu-Zn) (SOD1) and the Aldo-Keto-Reductase Family 1 Member b (AKR1B1) Are the Proteins Most Significantly Reduced by Cryopreservation. J Proteome Res 2021 May 7;20(5):2435-2446.
- Catalán J, Yánez-Ortiz I, Gacem S, Papas M, Bonet S, Rodríguez-Gil JE, Yeste M, Miró J. The Effects of Red Light on Mammalian Sperm Rely upon the Color of the Straw and the Medium Used. Animals (Basel) 2021 Jan 8;11(1).
- Catalán J, Papas M, Trujillo-Rojas L, Blanco-Prieto O, Bonilla-Correal S, Rodríguez-Gil JE, Miró J, Yeste M. Red LED Light Acts on the Mitochondrial Electron Chain of Donkey Sperm and Its Effects Depend on the Time of Exposure to Light. Front Cell Dev Biol 2020;8:588621.
- Blanco-Prieto O, Catalán J, Trujillo-Rojas L, Peña A, Rivera Del Álamo MM, Llavanera M, Bonet S, Fernández-Novell JM, Yeste M, Rodríguez-Gil JE. Red LED Light Acts on the Mitochondrial Electron Chain of Mammalian Sperm via Light-Time Exposure-Dependent Mechanisms. Cells 2020 Nov 26;9(12).
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