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RESEARCH ARTICLE
Contents Vol 64(11)

Effect of L-carnosine on frozen ram-semen quality evaluated by CASA and flow-cytometry

İbrahim Halil Güngör https://orcid.org/0000-0002-5250-1478 A * , Seyfettin Gür https://orcid.org/0000-0003-0096-2501 A and Edanur Güler Ekmen https://orcid.org/0000-0001-8473-7592 B
+ Author Affiliations
- Author Affiliations

A Department of Reproduction and Artificial Insemination, Fırat University, Faculty of Veterinary Medicine, Elazığ, Türkiye.

B Department of Physiology, Fırat University, Faculty of Veterinary Medicine, Elazığ, Türkiye.

* Correspondence to: ihgungor@firat.edu.tr

Handling Editor: Rodolfo Ungerfeld

Animal Production Science 64, AN24048 https://doi.org/10.1071/AN24048
Submitted: 16 February 2024  Accepted: 3 July 2024  Published: 29 July 2024

© 2024 The Author(s) (or their employer(s)). Published by CSIRO Publishing

Abstract

Context

Successful freezing of ram semen has not yet reached the desired levels. The main reason for this situation could be due to the fact that the spermatozoa of this species have a lipid composition different from that of other species.

Aims

The objective of the study was to evaluate the effect of different concentrations of L-carnosine added to the extender on ram semen after being frozen and thawed.

Methods

Semen was collected from six Akkaraman rams twice a week for a period of 3 weeks. Pooling was performed at each time. The semen were reconstituted with a pre-prepared tris + egg yolk solution and different amounts of L-carnosine to form experimental groups (Group 1: 1 mM, Group 2: 5 mM, Group 3: 10 mM, Group 4: 20 mM, Group 5: control) and were drawn into 0.25 mL mini straws. Subsequently, the samples were subjected to freezing by using an automated freezing device. Following the freezing process, the straws were placed in containers containing liquid nitrogen and thawed after 24 h.

Key results

After thawing, it was found that the samples containing 5 mM L-carnosine had superior results in all analyses. This concentration exhibited significantly higher percentages of progressive, total, and rapid sperm motility, live spermatozoa, high mitochondrial membrane potential rate, and higher GSH-Px concentrations. In addition, it was determined that 5 mM L-carnosine group protected the membrane integrity and significantly decreased the rate of abnormal spermatozoa, acrosomal damage rate, low mitochondrial membrane potential and apoptotic cell rate.

Conclusions

As a result, It was determined that adding 5 mM of L-carnosine to the semen extender during the freezing of ram samples would be beneficial for successful freezing.

Implications

The addition of 5 mM L-carnosine to ram-semen extenders ensures the freezability of the semen of this species; thus, this protocol could be used to perform artificial insemination with frozen ram semen.

Keywords: apoptosis, flow-cytometer, freeze-thawing, L-carnosine, oxidative stress, pH, ram, semen.

References

Bailey JL, Bilodeau J-F, Cormier N (2000) Semen cryopreservation in domestic animals: a damaging and capacitating phenomenon minireview. Journal of Andrology 21(1), 1-7.
| Crossref | Google Scholar | PubMed |

Barca A, Ippati S, Urso E, Vetrugno C, Storelli C, Maffia M, Romano A, Verri T (2019) Carnosine modulates the Sp1-Slc31a1/Ctr1 copper-sensing system and influences copper homeostasis in murine CNS-derived cells. American Journal of Physiology-Cell Physiology 316(2), C235-C245.
| Crossref | Google Scholar | PubMed |

Boldyrev A (2000) Problems and perspectives in studying the biological role of carnosine. Biochemistry 65(7), 751-756.
| Google Scholar | PubMed |

Boldyrev AA (2012) Carnosine: new concept for the function of an old molecule. Biochemistry (Moscow) 77(4), 313-326.
| Crossref | Google Scholar | PubMed |

Boldyrev AA, Aldini G, Derave W (2013) Physiology and pathophysiology of carnosine. Physiological Reviews 93, 1803-1845.
| Crossref | Google Scholar | PubMed |

Cho CH, Luk CT, Ogle CW (1991) The membrane-stabilizing action of zinc carnosine (Z-103) in stress-induced gastric ulceration in rats. Life Sciences 49(23), PL189-PL194.
| Crossref | Google Scholar | PubMed |

Davey CL (1960) The significance of carnosine and anserine in striated skeletal muscle. Archives of Biochemistry and Biophysics 89(2), 303-308.
| Crossref | Google Scholar |

de Agostini LJD, Angrimani DSR, Redondo RF, Brito MM, Rui BR, Kawai KV, Silva BCS, Mendes CM, Assumpção MEOD, Nichi M (2017) Deleterious effect of high carnosine concentrations in extenders during sperm cryopreservation in dogs. Journal of Veterinary Andrology 2(2), 60-67.
| Google Scholar |

Emamverdi M, Zhandi M, Shahneh AZ, Sharafi M, Akhlaghi A, Motlagh MK, Dadkhah F, Davachi ND (2014) Flow cytometric and microscopic evaluation of post-thawed ram semen cryopreserved in chemically defined home-made or commercial extenders. Animal Production Sciences 55(4), 551-558.
| Crossref | Google Scholar |

Güngör İH (2023) Koç Spermasının Kısa ve Uzun Süreli Saklanmasında L-karnosin Kullanımının Spermatolojik ve Flow-sitometrik Analizlerle Değerlendirilmesi. Doktora Tezi, Fırat Üniversitesi Veteriner Fakültesi, Elazığ, Türkiye. Available at https://tez.yok.gov.tr/UlusalTezMerkezi/tezSorguSonucYeni.jsp

Güngör İH, Kızıl M, Sönmez M, Gür S, Çambay Z, Yüce A, Türk G (2021) Effect of freeze–thawing process on lipid peroxidation, miRNAs, ion channels, apoptosis and global DNA methylation in ram spermatozoa. Reproduction, Fertility and Development 33(14), 747-759.
| Crossref | Google Scholar |

Goth L (1991) A simple method for determination of serum catalase activity and revision of reference range. Clinica Chimica Acta 196(2–3), 143-151.
| Crossref | Google Scholar |

Hipkiss AR (2000) Carnosine and protein carbonyl groups: a possible relationship. Biochemistry (Moscow) 65(7), 771-778.
| Google Scholar | PubMed |

Hipkiss AR, Michaelis J, Syrris P (1995) Non-enzymatic glycosylation of the dipeptide l-carnosine, a potential anti-protein-cross-linking agent. FEBS Letters 371(1), 81-85.
| Crossref | Google Scholar | PubMed |

Hipkiss AR, Preston JE, Himsworth DTM, et al. (1998) Pluripotent protective effects of carnosine, a naturally occurring dipeptide. Annals of the New York Academy of Sciences 854(1), 37-53.
| Crossref | Google Scholar |

Kawai GKV (2017) Effect of carnosine on the protection against cryoinjuries in semen of good and bad freezers’ stallions. Vol. 89, p. 170. Universidade de São Paulo, Faculdade de Medicina Veterinária e Zootecnia, São Paulo, Brazil.

Kohen R, Yamamoto Y, Cundy KC, Ames BN (1988) Antioxidant activity of carnosine, homocarnosine, and anserine present in muscle and brain. Proceedings of the National Academy of Sciences 85(9), 3175-3179.
| Crossref | Google Scholar |

Lawrence RA, Burk RF (1976) Glutathione peroxidase activity in selenium-deficient rat liver. Biochemical and Biophysical Research Communications 71(4), 952-958.
| Crossref | Google Scholar | PubMed |

Mozdzan M, Szemraj J, Rysz J, Nowak D (2005) Antioxidant properties of carnosine re-evaluated with oxidizing systems involving iron and copper ions. Basic & Clinical Pharmacology & Toxicology 96(5), 352-360.
| Crossref | Google Scholar | PubMed |

Placer ZA, Cushman LL, Johnson BC (1966) Estimation of product of lipid peroxidation (malonyl dialdehyde) in biochemical systems. Analytical Biochemistry 16(2), 359-364.
| Crossref | Google Scholar | PubMed |

Quinn PJ, Boldyrev AA, Formazuyk VE (1992) Carnosine: its properties, functions and potential therapeutic applications. Molecular Aspects of Medicine 13(5), 379-444.
| Crossref | Google Scholar |

Rocha CC, Kawai GK, de Agostini Losano JD, Angrimani DdSR, Rui BR, de Cássia Bicudo L, da Silva BdCS, Alonso MA, Mendes AM, Ortiz D’Avila Assumpção ME, Pereira RJG, Barnabe VH, Nichi M (2018) Carnosine as malondialdehyde scavenger in stallion seminal plasma and its role in sperm function and oxidative status. Theriogenology 119, 10-17.
| Crossref | Google Scholar | PubMed |

Sanchez-Partida LG, Setchell BP, Maxwell WMC (1997) Epididymal compounds and antioxidants in diluents for the frozen storage of ram spermatozoa. Reproduction, Fertility and Development 9(7), 689-696.
| Crossref | Google Scholar | PubMed |

Sedlak J, Lindsay RH (1968) Estimation of total, protein-bound, and nonprotein sulfhydryl groups in tissue with Ellman’s reagent. Analytical Biochemistry 25, 192-205.
| Crossref | Google Scholar | PubMed |

Türk G (2015) Reaktif oksijen türlerinin spermatozoon fonksiyonları üzerindeki fizyolojik ve patolojik etkileri. Turkiye Klinikleri Journal of Reproduction Artificial Insemination-Special Topics 1(3), 26-34.
| Google Scholar |

Türk G, Koca RH, Güngör İH, Dayan Cinkara S, Acısu TC, Erdem Erişir F, Arkalı G, Özer Kaya Ş, Kızıl M, Sönmez M, Gür S, Yılmaz Ö, Yüce A, Karatepe M (2022) Effect of hydrated C60 fullerene on lipid, vitamin and amino acid composition in frozen–thawed ram semen. Animal Reproduction Science 238, 106939.
| Crossref | Google Scholar | PubMed |

Wu G (2020) Important roles of dietary taurine, creatine, carnosine, anserine and 4-hydroxyproline in human nutrition and health. Amino Acids 52, 329-360.
| Crossref | Google Scholar | PubMed |

Xie Z, Baba SP, Sweeney BR, Barski OA (2013) Detoxification of aldehydes by histidine-containing dipeptides: from chemistry to clinical implications. Chemico-Biological Interactions 202(1–3), 288-297.
| Crossref | Google Scholar | PubMed |

Zhou S, Decker EA (1999) Ability of carnosine and other skeletal muscle components to quench unsaturated aldehydic lipid oxidation products. Journal of Agricultural and Food Chemistry 47(1), 51-55.
| Crossref | Google Scholar | PubMed |