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Intracellular calcium chelating agent (BAPTA-AM) aids stallion semen cooling and freezing-thawing.
Abstract: This study aimed to investigate the effects of different concentrations of 1,2-bis-(o-aminophenoxy)-ethane-N,N,N0 N0-tetraacetic acid, tetra-acetoxymethyl ester (BAPTA-AM), an intracellular calcium chelating agent, on stallion semen cooling and freezing-thawing. After collection, semen was extended (1:1 v/v) on a skim milk-based extender, centrifuged and resuspended at 400 million/ml into cooling or freezing extenders containing 0, 5, 25, 50, 100 and 200 μΜ BAPTA-AM. Motility parameters were assessed after cooling in Equitainer at 5°C for 12, 24, 48, 72 and 120 hr and after freezing-thawing. In addition, mitochondrial membrane potential, intracellular ATP, reactive oxygen species and malondialdehyde concentrations were measured in cryopreserved-thawed semen. Cooled stored (48 hr) semen containing 50 μΜ BAPTA-AM and control extender (0 μΜ BAPTA-AM) was used to assess fertility. Inclusion of 50 μΜ BAPTA-AM resulted in superior sperm motility parameters during cooled storage when compared to other groups (p < 0.05). Furthermore, semen cryopreserved in extender containing 50 μΜ BAPTA-AM showed increased intracellular ATP and mitochondrial membrane potential, whereas reactive oxygen species and malondialdehyde were increased after thawing for all groups (p < 0.05). Addition of 50 μΜ BAPTA-AM to cooling extender resulted in similar pregnancy rates to the control group (75% vs. 73.6%, respectively; p > 0.05). In conclusion, the addition of BAPTA-AM to semen extenders aided stallion semen cryopreservation in a dose-dependent manner. Furthermore, the cooling extender supplemented with 50 μΜ BAPTA-AM could be used to prolong the sperm motility during cooling without apparently compromising fertility. Field trials should be conducted to assess fertility of cryopreserved stallion semen with BAPTA-AM.
© 2018 Blackwell Verlag GmbH.
Publication Date: 2018-07-09 PubMed ID: 29984866DOI: 10.1111/rda.13245Google Scholar: Lookup The Equine Research Bank provides access to a large database of publicly available scientific literature. Inclusion in the Research Bank does not imply endorsement of study methods or findings by Mad Barn.
- Journal Article
Summary
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The research explores the influence of a calcium chelating agent, BAPTA-AM, on the cooling and freezing-thawing processes of stallion semen. The study found that adding BAPTA-AM can assist cryopreservation of stallion semen and potentially improve sperm motility during cooling without negatively affecting fertility.
Methods and Observations
- The researchers conducted experiments using stallion semen, which was first extended with a 1:1 ratio of a skim milk-based extender. The semen was then centrifuged and re-suspended in either cooling or freezing extenders with different concentrations of BAPTA-AM.
- The motility parameters were measured after cooling in an Equitainer at 5°C across various time points, and after the freezing-thawing process.
- Additionally, the scientists studied the mitochondrial membrane potential, intracellular ATP, reactive oxygen species, and malondialdehyde concentrations in the frozen-thawed semen.
- Two test groups were compared – semen treated with a control extender (0 µM BAPTA-AM) and semen treated with an extender containing 50 µM BAPTA-AM – to assess fertility after a 48-hour cooling period.
Results
- The results showed that the inclusion of 50 µM BAPTA-AM resulted in better sperm motility parameters during cooled storage compared to other groups.
- Also, semen frozen in extenders containing 50 µM BAPTA-AM exhibited increased intracellular ATP and mitochondrial membrane potential, while reactive oxygen species and malondialdehyde were observed to increase post-thawing across all groups.
- In terms of fertility, the addition of 50 µM BAPTA-AM to the cooling extender led to similar pregnancy rates as the control group (75% vs. 73.6%, respectively).
Conclusion
- Overall, the study concludes that the addition of BAPTA-AM to semen extenders supports the cryopreservation of stallion semen in a dose-dependent manner, specifically improving sperm motility during cooling without notably compromising fertility.
- However, the researchers recommend field trials for further investigation into the impact of BAPTA-AM on the fertility of cryopreserved stallion semen.
Cite This Article
APA
Wu S, Canisso IF, Yang W, Ul Haq I, Liu Q, Han Y, Zeng S.
(2018).
Intracellular calcium chelating agent (BAPTA-AM) aids stallion semen cooling and freezing-thawing.
Reprod Domest Anim, 53(5), 1235-1242.
https://doi.org/10.1111/rda.13245 Publication
Researcher Affiliations
- College of Animal Science and Technology, China Agricultural University, Beijing, China.
- Department of Veterinary Clinical Medicine, College of Veterinary Medicine, University of Illinois Urbana-Champaign, Urbana, Illinois.
- College of Animal Science and Technology, China Agricultural University, Beijing, China.
- Institute of Biotechnology and Genetic Engineering, University of Agriculture, Peshawar, Pakistan.
- College of Animal Science and Technology, China Agricultural University, Beijing, China.
- College of Animal Science and Technology, China Agricultural University, Beijing, China.
- College of Animal Science and Technology, China Agricultural University, Beijing, China.
- College of Animal Science and Technology, Yangzhou University, Yangzhou, China.
MeSH Terms
- Animals
- Calcium Chelating Agents / pharmacology
- Cryopreservation / veterinary
- Cryoprotective Agents / pharmacology
- Egtazic Acid / analogs & derivatives
- Egtazic Acid / pharmacology
- Female
- Horses
- Male
- Membrane Potential, Mitochondrial
- Pregnancy
- Pregnancy Rate
- Reactive Oxygen Species / analysis
- Semen Preservation / veterinary
- Sperm Motility / drug effects
- Spermatozoa / drug effects
Grant Funding
- 20130008130001 / Specialized Research Fund for the Doctoral Program for Higher Education
- 31470077 / Natural Science Foundation of China
Citations
This article has been cited 4 times.- Zhou Q, Li T, Fang G, Pang Y, Wang X. Bioactive Molecules against Rheumatoid Arthritis by Suppressing Pyroptosis. Pharmaceuticals (Basel) 2023 Jul 3;16(7).
- Lv X, Chen Z, Zheng M, Bai R, Zhang L, Zhang X, Duan B, Zhao Y, Yin L, Fan B, Cui K, Xu T. The interaction between free Ca(2+) in host cells and invasion of E. tenella. Parasitol Res 2022 Mar;121(3):965-972.
- Najafi A, Asadi E, Benson JD. Comparative effects of a calcium chelator (BAPTA-AM) and melatonin on cryopreservation-induced oxidative stress and damage in ovarian tissue. Sci Rep 2023 Dec 21;13(1):22911.
- Sushadi PS, Kuwabara M, Maung EEW, Mohamad Mohtar MS, Sakamoto K, Selvaraj V, Asano A. Arresting calcium-regulated sperm metabolic dynamics enables prolonged fertility in poultry liquid semen storage. Sci Rep 2023 Dec 8;13(1):21775.
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