Dimethyl formamide improves the postthaw characteristics of sex-sorted and nonsorted stallion sperm.
Abstract: Cryopreserved, sex-sorted stallion sperm has been shown to have poor fertility. During this study, the effects of cryoprotectant (glycerol [GLY] and dimethyl formamide [DMF]), cryoprotectant equilibration time (0, 30, 60, 90, or 120 minutes), and cryoprotectant concentration (2%, 3%, or 4% vol/vol) on stored sex-sorted and stored nonsorted stallion sperm were evaluated. Total motility, viability, and DNA integrity (determined using sperm chromatin structure assay) of sperm were assessed after thawing. Equilibration for 90 minutes improved total motility (33.8%) compared with 0 (28.5%) or 120 minutes (29.8%; P < 0.05), though viability was higher after 120 minutes (33.1%) compared with 0 (30.5%) or 30 minutes (31.0%; P < 0.01). The viability of nonsorted sperm decreased as cryoprotectant concentration increased (P < 0.001), and total motility of nonsorted sperm was higher when DMF alone was used (15.8%, 16.6%, and 24.0% for GLY, GLY and DMF, and DMF respectively; P < 0.001). Sex sorting was detrimental to the postthaw quality of sperm; at 45 minutes after thawing, total motility of nonsorted sperm was higher than that of sex-sorted sperm (37.4% vs. 5.6%; P < 0.001), the viability of sex-sorted sperm was lower than that of nonsorted sperm (12.4% vs. 30.0%; P < 0.001, averaged over postthaw time), and sex-sorted sperm had higher detectable DNA fragmentation index (DFI) (63.6% vs. 11.3%, P < 0.001) and mean DFI (285.1 vs. 211.3, P < 0.001) than nonsorted sperm. The viability of sex-sorted sperm was improved by GLY and DMF or DMF compared with GLY (22.6%, 25.3%, and 19.3%, respectively; P < 0.05), and the DNA integrity of sex-sorted sperm was improved by the use of DMF compared with GLY (detectable DFI, 60.2 vs. 66.8, P < 0.05; and mean DFI, 280.9 vs. 289.2, P < 0.05, respectively). In conclusion, postthaw characteristics of stored sex-sorted and stored nonsorted stallion sperm were improved by the use of DMF as a cryoprotectant, though the parameters to benefit differed between sorted and nonsorted sperm.
Copyright © 2013 Elsevier Inc. All rights reserved.
Publication Date: 2013-03-01 PubMed ID: 23453787DOI: 10.1016/j.theriogenology.2013.01.013Google Scholar: Lookup
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- Journal Article
- Research Support
- Non-U.S. Gov't
Summary
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The research article is a scientific study investigating the effects of dimethyl formamide (DMF), a cryoprotectant, on the post-thaw characteristics of cryopreserved, sex-sorted and nonsorted stallion sperm.
Introduction and Method
- The study focused on improving the fertility of cryopreserved, sex-sorted stallion sperm, which has been observed to have poor fertility.
- The research evaluated different variables, including cryoprotectants used (glycerol [GLY] and dimethyl formamide [DMF]), the equilibration time of cryoprotectant (0, 30, 60, 90, or 120 minutes), and the concentration of cryoprotectant (2%, 3%, or 4% vol/vol).
- After freezing and subsequent thawing of the stallion sperm, three parameters were measured: total motility, viability, and DNA integrity, the latter determined using a sperm chromatin structure assay.
Results
- The results displayed a direct relationship between equilibration time and sperm viability and motility. The sperm had the highest motility after 90 minutes of equilibration, however, viability was highest after 120 minutes.
- Cryoprotectant concentration had a negative effect on the viability of nonsorted sperm. As cryoprotectant concentration increased, sperm viability decreased.
- The use of DMF as a cryoprotectant produced a higher total motility of nonsorted sperm compared to using GLY alone or a combination of GLY and DMF.
- Sex-sorting had a negative impact on sperm post-thaw quality. Measurements taken 45 minutes post-thaw showed that nonsorted sperm had a higher total motility and viability than sex-sorted sperm. Furthermore, sex-sorted sperm had a higher detectable DNA fragmentation index (DFI) and mean DFI than nonsorted sperm, indicating a higher level of DNA damage.
- However, adding DMF to the GLY or using DMF alone improved the viability of sex-sorted sperm compared to using GLY alone. Furthermore, DMF also improved the DNA integrity of sex-sorted sperm compared to GLY.
Conclusion
- The findings show that using DMF as a cryoprotectant improved the post-thaw characteristics of both sex-sorted and nonsorted stallion sperm.
- The benefits differed between sorted and nonsorted sperm. For nonsorted sperm, DMF alone improved the total motility, while for sex-sorted sperm, a combination of DMF and GLY or DMF alone improved viability and DNA integrity.
Cite This Article
APA
Gibb Z, Morris LH, Maxwell WM, Grupen CG.
(2013).
Dimethyl formamide improves the postthaw characteristics of sex-sorted and nonsorted stallion sperm.
Theriogenology, 79(7), 1027-1033.
https://doi.org/10.1016/j.theriogenology.2013.01.013 Publication
Researcher Affiliations
- Faculty of Veterinary Science, The University of Sydney, Camperdown, New South Wales, Australia. zamiragibb@gmail.com
MeSH Terms
- Animals
- Cryopreservation / methods
- Cryopreservation / veterinary
- Cryoprotective Agents / pharmacology
- DNA Fragmentation
- Dimethylformamide / pharmacology
- Flow Cytometry / methods
- Flow Cytometry / veterinary
- Horses
- Male
- Semen Preservation / methods
- Semen Preservation / veterinary
- Sex Preselection / methods
- Sex Preselection / veterinary
- Spermatozoa / cytology
- Spermatozoa / drug effects
Citations
This article has been cited 2 times.- Snoeck PPDN, Pessoa THO, Pereira MGS, Bastos ICL, de Melo MIV. Can we use LDL instead of egg yolk in BotuCrio® extender to cryopreserve sperm from the Mangalarga Marchador stallion?. Anim Reprod 2019 Oct 23;16(2):340-347.
- Ortiz-Rodriguez JM, Balao da Silva C, Masot J, Redondo E, Gazquez A, Tapia JA, Gil C, Ortega-Ferrusola C, Peña FJ. Rosiglitazone in the thawing medium improves mitochondrial function in stallion spermatozoa through regulating Akt phosphorylation and reduction of caspase 3. PLoS One 2019;14(7):e0211994.
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