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Scientific reports2021; 11(1); 14076; doi: 10.1038/s41598-021-93569-y

Drying and temperature induced conformational changes of nucleic acids and stallion sperm chromatin in trehalose preservation formulations.

Abstract: Even though dried sperm is not viable, it can be used for fertilization as long as its chromatin remains intact. In this study, we investigated drying- and temperature-induced conformational changes of nucleic acids and stallion sperm chromatin. Sperm was diluted in preservation formulations with and without sugar/albumin and subjected to convective drying at elevated temperatures on glass substrates. Accumulation of reactive oxygen species was studied during storage at different temperatures, and the sperm chromatin structure assay was used to assess DNA damage. Fourier transform infrared spectroscopy was used to identify dehydration and storage induced conformational changes in isolated DNA and sperm chromatin. Furthermore, hydrogen bonding in the preservation solutions associated with storage stability were investigated. Reactive oxygen species and DNA damage in dried sperm samples were found to accumulate with increasing storage temperature and storage duration. Non-reducing disaccharides (i.e., trehalose, sucrose) and albumin counteracted oxidative stress and preserved sperm chromatin during dried storage, whereas glucose increased DNA damage during storage. When sperm was dried in the presence of trehalose and albumin, no spectral changes were detected during storage at refrigeration temperatures, whereas under accelerated aging conditions, i.e., storage at 37 °C, spectral changes were detected indicating alterations in sperm chromatin structure.
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Publication Date: 2021-07-07 PubMed ID: 34234244PubMed Central: PMC8263733DOI: 10.1038/s41598-021-93569-yGoogle Scholar: Lookup
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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.

This study investigates the impacts of drying and temperature on nucleic acids and sperm chromatin, emphasizing the use of trehalose preservation formulations. It finds that both factors can damage DNA but sugar-based formulas can mitigate these impacts.

Research Methodology

  • The researchers collected stallion sperm, then diluted the samples in preservation solutions both with and without sugar and albumin.
  • This diluted sperm was then subjected to convective drying using elevated temperatures on the given glass substrates.
  • Different storage temperatures were used to monitor the collection of reactive oxygen species which are often produced under stressful conditions and can be harmful to cellular functions.
  • The research used a sperm chromatin structure assay to study any potential DNA damage that occurred.
  • Further, Fourier transform infrared spectroscopy was used to identify changes in conformation – the three-dimensional shape of a protein that is crucial for its function – induced by drying and storage.

Key Findings

  • They recorded an increase in reactive oxygen species and DNA damage in the dried sperm samples as storage temperature and duration increased.
  • The team discovered that non-reducing disaccharides like trehalose and sucrose, as well as albumin, were successful in counteracting oxidative stress and preserving sperm chromatin during the dried storage process. In contrast, they found that glucose increased DNA damage over the storage period.
  • When the sperm was dried with trehalose and albumin present, no detectable spectral changes were observed during storage at refrigeration temperatures.
  • However, under accelerated aging conditions, such as storage at 37° C, they noted spectral changes that indicated shifts in sperm chromatin structure.

Study Significance

This research provides an important look at how sperm can be preserved, which has implications for reproductive science and fertility treatments. The findings indicate that it’s possible to design preservation solutions that protect sperm chromatin from oxidative stress and DNA damage, with potential for improved success in assisted fertility treatments. However, it also flags that glucose, often used in these solutions, can in fact increase DNA damage during storage.

Cite This Article

APA
Brogna R, Fan J, Sieme H, Wolkers WF, Oldenhof H. (2021). Drying and temperature induced conformational changes of nucleic acids and stallion sperm chromatin in trehalose preservation formulations. Sci Rep, 11(1), 14076. https://doi.org/10.1038/s41598-021-93569-y

Publication

Scientific reports
ISSN: 2045-2322
NlmUniqueID: 101563288
Country: England
Language: English
Volume: 11
Issue: 1
Pages: 14076
PII: 14076

Researcher Affiliations

Brogna, Raffaele
  • Biostabilization Laboratory, Lower Saxony Centre for Biomedical Engineering, Implant Research and Development, University of Veterinary Medicine Hannover, Hannover, Germany.
  • Unit for Reproductive Medicine, Clinic for Horses, University of Veterinary Medicine Hannover, Bünteweg 15, 30559, Hannover, Germany.
Fan, Juezhu
  • Biostabilization Laboratory, Lower Saxony Centre for Biomedical Engineering, Implant Research and Development, University of Veterinary Medicine Hannover, Hannover, Germany.
Sieme, Harald
  • Unit for Reproductive Medicine, Clinic for Horses, University of Veterinary Medicine Hannover, Bünteweg 15, 30559, Hannover, Germany.
Wolkers, Willem F
  • Biostabilization Laboratory, Lower Saxony Centre for Biomedical Engineering, Implant Research and Development, University of Veterinary Medicine Hannover, Hannover, Germany.
  • Unit for Reproductive Medicine, Clinic for Horses, University of Veterinary Medicine Hannover, Bünteweg 15, 30559, Hannover, Germany.
Oldenhof, Harriëtte
  • Unit for Reproductive Medicine, Clinic for Horses, University of Veterinary Medicine Hannover, Bünteweg 15, 30559, Hannover, Germany. harriette.oldenhof@tiho-hannover.de.

MeSH Terms

  • Animals
  • Chromatin
  • DNA Damage
  • Freeze Drying
  • Male
  • Nucleic Acid Conformation
  • Nucleic Acids / chemistry
  • Reactive Oxygen Species
  • Semen Preservation
  • Spectroscopy, Fourier Transform Infrared
  • Spermatozoa / metabolism
  • Temperature

Conflict of Interest Statement

The authors declare no competing interests.

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Citations

This article has been cited 2 times.
  1. Comizzoli P, He X, Lee PC. Long-term preservation of germ cells and gonadal tissues at ambient temperatures. Reprod Fertil 2022 Apr 1;3(2):R42-R50.
    doi: 10.1530/RAF-22-0008pubmed: 35514540google scholar: lookup
  2. Lee PC, Zahmel J, Jewgenow K, Comizzoli P. Desiccated cat spermatozoa retain DNA integrity and developmental potential after prolonged storage and shipping at non-cryogenic temperatures. J Assist Reprod Genet 2022 Jan;39(1):141-151.
    doi: 10.1007/s10815-021-02337-4pubmed: 34609666google scholar: lookup
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