Impact of Cryopreservation on Caspase-3 Gene Expression, Cell Viability, and Sperm Motility in Bull Testicular Tissue and Epididymal Sperm

Year 2025, Volume: 10 Issue: 2, 496 - 503, 31.08.2025

Cumali Kaya , Burcu Esin , Cansu Can , Eda Turgut Uğurtay , Mesut Çevik

https://doi.org/10.24880/meditvetj.1633870

Abstract

This study investigated the effects of cryopreservation of testicular tissue and epididymal sperm in bulls on spermatological parameters, cell viability, and caspase-3 gene expression, a marker of apoptosis. Epididymal sperm and testicular tissue pieces obtained from testes collected from bulls (n: 15) were cryopreserved and stored in liquid nitrogen (-196°C). Epididymal sperm were frozen in liquid nitrogen vapor using the straw method, while testicular tissue pieces were frozen in cryotubes using a slow freezing protocol with Dimethylsulfoxide (DMSO) and Ethylene Glycol (EG) as cryoprotectants. Fresh semen exhibited significantly higher motility, progressive motility, kinematic parameters, and viability values compared to those after thawing (P < 0.05). Higher cell viability was achieved in DMSO-cryopreserved testicular tissues (53.72 ± 6.36) compared to EG-cryopreserved testicular tissues (41.83 ± 7.22) (P < 0.05). Caspase-3 gene expression was determined to be 42.66 ± 98.26-fold increased in DMSO-cryopreserved tissues and 65.05 ± 106.03 fold increased in EG-cryopreserved tissues after thawing. In epididymal sperm, gene expression levels increased by an average of 12.22 ± 34.60 fold compared to fresh samples. These findings suggest that cryopreservation affects cellular integrity and triggers apoptotic pathways and repair mechanisms, highlighting the need to optimize protocols to reduce cryo-induced damage and increase post-thaw viability.

Keywords

cryopreservation , caspase-3 , epididiymal sperm , gene expression , testicular tissue

Kriyoprezervasyonun Boğa Testis Dokusu ve Epididimal Spermasındaki Kaspaz-3 Gen Ekspresyonu, Hücre Canlılığı ve Sperm Hareketliliği Üzerindeki Etkisi

Abstract

Bu çalışmada, boğalarda testis dokusu ve epididimal sperma kriyoprezervasyonunun spermatolojik parametreler, hücre canlılığı ve apoptozis belirteci olan kaspaz-3 gen ekspresyonu üzerine etkileri araştırılmıştır. Boğalardan toplanan testislerden elde edilen epididimal sperma ve testis dokusu parçaları (n: 15) kriyoprezervasyon sonrasında sıvı nitrojende (-196 °C) saklanmıştır. Epididimal sperma payet yöntemi kullanılarak sıvı nitrojen buharında dondurulurken, testis dokusu parçaları kriyoprotektan olarak Dimetilsülfoksit (DMSO) ve Etilen Glikol (EG) içeren kriyotüplerde yavaş dondurma protokolü kullanılarak dondurulmuştur. Taze sperma, çözüm sonrası örneklere kıyasla önemli ölçüde daha yüksek motilite, progresif motilite, kinematik parametreler ve canlılık değerleri göstermiştir (P < 0,05). DMSO ile kriyoprezerve edilen testis dokularında (53.72 ± 6.36), EG ile kriyoprezerve edilen testis dokularına (41.83 ± 7.22) kıyasla daha yüksek hücre canlılığı elde edildi (P < 0.05). Kaspaz-3 gen ekspresyonunun çözüm sonrası DMSO grubundaki dokularda 42.66 ± 98.26 kat, EG grubundaki dokularda ise 65.05 ± 106.03 kat arttığı belirlendi. Epididimal spermade gen ekspresyon düzeyleri taze spermaya kıyasla çözüm sonrası spermada ortalama 12.22 ± 34.60 kat arttı. Bu bulgular kriyoprezervasyonun hücre bütünlüğünü etkilediğini ve apoptotik yolları ve onarım mekanizmalarını tetiklediğini düşündürerek, kriyo hasarı azaltmak ve çözüm sonrası canlılığı artırmak için protokolleri optimize etme ihtiyacını vurgulamaktadır.

Keywords

kriyoprezervasyon , kaspaz-3 , epididimal sperma , gen ekspresyonu , testis dokusu

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