Boar Sperm Cryopreservation Improvement Using Semen Extender Modification by Dextran and Pentaisomaltose
The long-term storage of boar sperm presents an ongoing challenge, and the modification of the cryoprotective compounds in semen extenders is crucial for improving cryopreservation’s success rate. The aim of our study was to reduce the percentage of glycerol in the extender by elimination or substit...
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| Format: | Article |
| Language: | English |
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MDPI AG
2022-03-01
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| Series: | Animals |
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| Online Access: | https://www.mdpi.com/2076-2615/12/7/868 |
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| author | Ondrej Simonik Filipa Bubenickova Lucie Tumova Michaela Frolikova Vishma Pratap Sur Jan Beran Katerina Havlikova Lenka Hackerova Daniela Spevakova Katerina Komrskova Pavla Postlerova |
| author_facet | Ondrej Simonik Filipa Bubenickova Lucie Tumova Michaela Frolikova Vishma Pratap Sur Jan Beran Katerina Havlikova Lenka Hackerova Daniela Spevakova Katerina Komrskova Pavla Postlerova |
| author_sort | Ondrej Simonik |
| collection | DOAJ |
| description | The long-term storage of boar sperm presents an ongoing challenge, and the modification of the cryoprotective compounds in semen extenders is crucial for improving cryopreservation’s success rate. The aim of our study was to reduce the percentage of glycerol in the extender by elimination or substitution with biocompatible, non-toxic polysaccharides. For boar semen extender improvement, we tested a novel modification with the polysaccharides dextran and pentaisomaltose in combination with unique in silico predictive modeling. We targeted the analysis of in vitro qualitative sperm parameters such as motility, viability, mitochondrial activity, acrosome integrity, and DNA integrity. Non-penetrating polysaccharide-based cryoprotective agents interact with sperm surface proteins such as spermadhesins, which are recognized as fertility markers of boar sperm quality. The in silico docking study showed a moderate binding affinity of dextran and pentaisomaltose toward one specific spermadhesin known as AWN, which is located in the sperm plasma membrane. Pentaisomaltose formed a hydrophobic pocket for the AWN protein, and the higher energy of this protein–ligand complex compared with dextran was calculated. In addition, the root mean square deviation (RMSD) analysis for the molecular dynamics (MD) of both polysaccharides and AWN simulation suggests their interaction was highly stable. The in silico results were supported by in vitro experiments. In the experimental groups where glycerol was partially or entirely substituted, the use of pentaisomaltose resulted in improved sperm mitochondrial activity and DNA integrity after thawing when compared with dextran. In this paper, we demonstrate that pentaisomaltose, previously used for cryopreservation in hematopoietic stem cells, represents a promising compound for the elimination or reduction of glycerol in extenders for boar semen cryopreservation. This novel approach, using in silico computer prediction and in vitro testing, represents a promising technique to help identify new cryoprotectants for use in animal breeding or genetic resource programs. |
| first_indexed | 2024-03-09T12:11:56Z |
| format | Article |
| id | doaj.art-980b7d14772747a0be5c3e95efdcdf40 |
| institution | Directory Open Access Journal |
| issn | 2076-2615 |
| language | English |
| last_indexed | 2024-03-09T12:11:56Z |
| publishDate | 2022-03-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Animals |
| spelling | doaj.art-980b7d14772747a0be5c3e95efdcdf402023-11-30T22:51:30ZengMDPI AGAnimals2076-26152022-03-0112786810.3390/ani12070868Boar Sperm Cryopreservation Improvement Using Semen Extender Modification by Dextran and PentaisomaltoseOndrej Simonik0Filipa Bubenickova1Lucie Tumova2Michaela Frolikova3Vishma Pratap Sur4Jan Beran5Katerina Havlikova6Lenka Hackerova7Daniela Spevakova8Katerina Komrskova9Pavla Postlerova10Laboratory of Reproductive Biology, Institute of Biotechnology of the Czech Academy of Sciences, BIOCEV, Prumyslova 595, 25250 Vestec, Czech RepublicDepartment of Veterinary Sciences, Faculty of Agrobiology, Food and Natural Resources, University of Life Sciences Prague, Kamycka 129, 16500 Prague, Czech RepublicDepartment of Veterinary Sciences, Faculty of Agrobiology, Food and Natural Resources, University of Life Sciences Prague, Kamycka 129, 16500 Prague, Czech RepublicLaboratory of Reproductive Biology, Institute of Biotechnology of the Czech Academy of Sciences, BIOCEV, Prumyslova 595, 25250 Vestec, Czech RepublicLaboratory of Reproductive Biology, Institute of Biotechnology of the Czech Academy of Sciences, BIOCEV, Prumyslova 595, 25250 Vestec, Czech RepublicDepartment of Zootechnical Sciences, Faculty of Agriculture, University of South Bohemia in Ceske Budejovice, Studentska 1668, 37005 Ceske Budejovice, Czech RepublicDepartment of Veterinary Sciences, Faculty of Agrobiology, Food and Natural Resources, University of Life Sciences Prague, Kamycka 129, 16500 Prague, Czech RepublicDepartment of Veterinary Sciences, Faculty of Agrobiology, Food and Natural Resources, University of Life Sciences Prague, Kamycka 129, 16500 Prague, Czech RepublicLaboratory of Reproductive Biology, Institute of Biotechnology of the Czech Academy of Sciences, BIOCEV, Prumyslova 595, 25250 Vestec, Czech RepublicLaboratory of Reproductive Biology, Institute of Biotechnology of the Czech Academy of Sciences, BIOCEV, Prumyslova 595, 25250 Vestec, Czech RepublicLaboratory of Reproductive Biology, Institute of Biotechnology of the Czech Academy of Sciences, BIOCEV, Prumyslova 595, 25250 Vestec, Czech RepublicThe long-term storage of boar sperm presents an ongoing challenge, and the modification of the cryoprotective compounds in semen extenders is crucial for improving cryopreservation’s success rate. The aim of our study was to reduce the percentage of glycerol in the extender by elimination or substitution with biocompatible, non-toxic polysaccharides. For boar semen extender improvement, we tested a novel modification with the polysaccharides dextran and pentaisomaltose in combination with unique in silico predictive modeling. We targeted the analysis of in vitro qualitative sperm parameters such as motility, viability, mitochondrial activity, acrosome integrity, and DNA integrity. Non-penetrating polysaccharide-based cryoprotective agents interact with sperm surface proteins such as spermadhesins, which are recognized as fertility markers of boar sperm quality. The in silico docking study showed a moderate binding affinity of dextran and pentaisomaltose toward one specific spermadhesin known as AWN, which is located in the sperm plasma membrane. Pentaisomaltose formed a hydrophobic pocket for the AWN protein, and the higher energy of this protein–ligand complex compared with dextran was calculated. In addition, the root mean square deviation (RMSD) analysis for the molecular dynamics (MD) of both polysaccharides and AWN simulation suggests their interaction was highly stable. The in silico results were supported by in vitro experiments. In the experimental groups where glycerol was partially or entirely substituted, the use of pentaisomaltose resulted in improved sperm mitochondrial activity and DNA integrity after thawing when compared with dextran. In this paper, we demonstrate that pentaisomaltose, previously used for cryopreservation in hematopoietic stem cells, represents a promising compound for the elimination or reduction of glycerol in extenders for boar semen cryopreservation. This novel approach, using in silico computer prediction and in vitro testing, represents a promising technique to help identify new cryoprotectants for use in animal breeding or genetic resource programs.https://www.mdpi.com/2076-2615/12/7/868cryopreservationboar spermdextranpentaisomaltosepolysaccharidereproduction |
| spellingShingle | Ondrej Simonik Filipa Bubenickova Lucie Tumova Michaela Frolikova Vishma Pratap Sur Jan Beran Katerina Havlikova Lenka Hackerova Daniela Spevakova Katerina Komrskova Pavla Postlerova Boar Sperm Cryopreservation Improvement Using Semen Extender Modification by Dextran and Pentaisomaltose Animals cryopreservation boar sperm dextran pentaisomaltose polysaccharide reproduction |
| title | Boar Sperm Cryopreservation Improvement Using Semen Extender Modification by Dextran and Pentaisomaltose |
| title_full | Boar Sperm Cryopreservation Improvement Using Semen Extender Modification by Dextran and Pentaisomaltose |
| title_fullStr | Boar Sperm Cryopreservation Improvement Using Semen Extender Modification by Dextran and Pentaisomaltose |
| title_full_unstemmed | Boar Sperm Cryopreservation Improvement Using Semen Extender Modification by Dextran and Pentaisomaltose |
| title_short | Boar Sperm Cryopreservation Improvement Using Semen Extender Modification by Dextran and Pentaisomaltose |
| title_sort | boar sperm cryopreservation improvement using semen extender modification by dextran and pentaisomaltose |
| topic | cryopreservation boar sperm dextran pentaisomaltose polysaccharide reproduction |
| url | https://www.mdpi.com/2076-2615/12/7/868 |
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