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Andrology2021; 9(5); 1631-1651; doi: 10.1111/andr.13036

Factors affecting intracellular calcium influx in response to calcium ionophore A23187 in equine sperm.

Abstract: Exposure to the calcium ionophore A23187 may present a "universal" sperm treatment for IVF, as it bypasses capacitation pathways. However, success in utilizing A23187 is variable, especially in equine spermatozoa. Notably, albumin is used during A23187 treatment but paradoxically is thought to suppress A23187 action. Essentially no critical data are available on the effects of A23187 and albumin concentrations, ratios, or addition protocols on changes in intracellular calcium ([Ca] ) in any cell type. To determine factors that affect the action of A23187 on [Ca] in equine and murine spermatozoa. Spermatozoa were loaded with Fluo-4 and changes in fluorescence after A23187 treatment were measured under various conditions using a microplate reader. Concentrations of bovine serum albumin (BSA) and A23187, type of BSA, makeup of A23187 stock solutions (i.e., 1° stock (DMSO) or 2° stock made with medium, water or DMSO), order of addition of spermatozoa and A23187, incubation of media before sperm addition, species of spermatozoa, and time of addition of BSA all affected [Ca] in response to A23187 treatment. In equine spermatozoa already exposed to 10 µM A23187, addition of BSA to 33 mg/ml to "quench" the A23187 did not affect [Ca] . When this concentration of BSA was added to spermatozoa exposed to 1 µM A23187, [Ca] in murine spermatozoa returned to baseline, however, equine spermatozoa continued to exhibit increased [Ca] . Addition of BSA to 33 mg/ml to media containing 1 µM A23187, prior to addition of spermatozoa, completely inhibited change in [Ca] in both murine and equine spermatozoa. These results represent some of the first critical data on the effects of albumin and other procedural factors on A23187-induced changes in [Ca] in any cell type. Our findings help to explain the variability in reported response of spermatozoa to A23187 among species and among laboratories.
© 2021 American Society of Andrology and European Academy of Andrology.
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Publication Date: 2021-06-23 PubMed ID: 33998170DOI: 10.1111/andr.13036Google Scholar: Lookup
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  • Non-U.S. Gov't

Summary

This research summary has been generated with artificial intelligence and may contain errors and omissions. Refer to the original study to confirm details provided. Submit correction.

The research investigates how the calcium ionophore A23187, used in in-vitro fertilization (IVF), affects calcium levels inside equine and murine (mouse) sperm cells, and the role of bovine serum albumin (BSA) in this process.

Overview of Study and Plan

  • The researchers primarily aimed at determining the factors affecting the interaction of the calcium ionophore A23187 with intracellular calcium concentration in equine and murine spermatozoa.
  • The calcium ionophore A23187 is used in IVF as it is believed to bypass the capacitation pathways, however, its effectiveness varies, particularly in equine spermatozoa.
  • The team used the Fluo-4 compound to measure changes in calcium across different scenarios using a microplate reader, this validates the intracellular calcium (also referred to as [Ca]) in response to A23187 treatment.

Treatment Scenarios and Observations

  • Multiple elements including the A23187 and BSA concentrations, type of BSA, composition of A23187 stock solutions, order of spermatozoa and A23187 addition, incubation of media before sperm addition, spermatozoa species, and BSA addition timing all played a role in changing [Ca] due to A23187 treatment.
  • When BSA was added at a concentration of 33 mg/ml to equine sperm already treated with 10µM A23187, it did not affect [Ca]. This suggests that at high A23187 concentrations, BSA might not suppress its action.
  • The same BSA concentration, when added to sperm exposed to a low amount of A23187 (1µM), caused [Ca] in murine sperm to return to baseline, while it remained high in equine sperm.
  • When the 33 mg/ml BSA was added to the media containing 1µM A23187 prior to the addition of spermatozoa, it inhibited [Ca] change in both murine and equine spermatozoa. This hints at the preventive role of BSA when present prior to A23187 interaction.

Significance and Conclusion

  • These results represent early critical data on how albumin and other procedural factors affect A23187-induced changes in [Ca] in a cell.
  • This research may be important for optimizing IVF treatments, particularly in equine species, contributing to greater understanding of interspecies and inter-laboratory differences in [Ca] responses to A23187.

Cite This Article

APA
Sampaio B, Ortiz I, Resende H, Felix M, Varner D, Hinrichs K. (2021). Factors affecting intracellular calcium influx in response to calcium ionophore A23187 in equine sperm. Andrology, 9(5), 1631-1651. https://doi.org/10.1111/andr.13036

Publication

Andrology
ISSN: 2047-2927
NlmUniqueID: 101585129
Country: England
Language: English
Volume: 9
Issue: 5
Pages: 1631-1651

Researcher Affiliations

Sampaio, Breno
  • College of Veterinary Medicine & Biomedical Sciences, Texas A&M University, College Station, TX, USA.
Ortiz, Isabel
  • College of Veterinary Medicine & Biomedical Sciences, Texas A&M University, College Station, TX, USA.
Resende, Hélène
  • College of Veterinary Medicine & Biomedical Sciences, Texas A&M University, College Station, TX, USA.
Felix, Matheus
  • College of Veterinary Medicine & Biomedical Sciences, Texas A&M University, College Station, TX, USA.
Varner, Dickson
  • College of Veterinary Medicine & Biomedical Sciences, Texas A&M University, College Station, TX, USA.
Hinrichs, Katrin
  • College of Veterinary Medicine & Biomedical Sciences, Texas A&M University, College Station, TX, USA.

MeSH Terms

  • Animals
  • Calcimycin / pharmacology
  • Calcium / metabolism
  • Calcium Ionophores / pharmacology
  • Horses
  • Male
  • Mice
  • Serum Albumin, Bovine / metabolism
  • Spermatozoa / drug effects

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Citations

This article has been cited 3 times.
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