Effect of freezing rate on acrosome and chromatin integrity in ram semen

ZIK, Berrin; ÜSTÜNER, BURCU; GÜNAY, Ülgen; TÜTÜNCÜ, ŞERİFE; DOĞAN, İbrahim; SAĞIRKAYA, Hakan; OZGUDEN, G. Cansel; NUR, ZEKARIYA

Effect of freezing rate on acrosome and chromatin integrity in ram semen

Zekariya NUR, (Uludağ Üniversitesi, Veteriner Fakültesi, Dölerme ve Suni Tohumlama Anabilim Dalı, Bursa, Türkiye)

Berrin ZIK, (Uludağ Üniversitesi, Veteriner Fakültesi, Histoloji ve Embriyoloji Anabilim Dalı, Bursa, Türkiye)

BURCU ÜSTÜNER, (Uludağ Üniversitesi, Veteriner Fakültesi, Dölerme ve Suni Tohumlama Anabilim Dalı, Bursa, Türkiye)

ŞERİFE TÜTÜNCÜ, (Uludağ Üniversitesi, Veteriner Fakültesi, Histoloji ve Embriyoloji Anabilim Dalı, Bursa, Türkiye)

Hakan SAĞIRKAYA, (Uludağ Üniversitesi, Veteriner Fakültesi, Dölerme ve Suni Tohumlama Anabilim Dalı, Bursa, Türkiye)

G. Cansel OZGUDEN, (Uludağ Üniversitesi, Veteriner Fakültesi, Histoloji ve Embriyoloji Anabilim Dalı, Bursa, Türkiye)

Ülgen GÜNAY, (Uludağ Üniversitesi, Veteriner Fakültesi, Dölerme ve Suni Tohumlama Anabilim Dalı, Bursa, Türkiye)

İbrahim DOĞAN (Uludağ Üniversitesi, Veteriner Fakültesi, Dölerme ve Suni Tohumlama Anabilim Dalı, Bursa, Türkiye)

Ankara Üniversitesi Veteriner Fakültesi Dergisi

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Yıl: 2011 Cilt: 58 Sayı: 4 Sayfa Aralığı: 267 - 272 Metin Dili: Türkçe İndeks Tarihi: 29-07-2022

Effect of freezing rate on acrosome and chromatin integrity in ram semen

Öz:

Bu çalışma, farklı dondurma hızlarının, eritme sonrası motilite, akrozom bütünlüğü, kromatin yapısı ve apoptozis üzerine olan etkilerini araştırmak amacıyla gerçekleştirildi. Toplanan sperma örnekleri 30ºC’de 1/5 oranında (sperma/sulandırıcı) Bioxel® (IMV, Fransa) ile sulandırıldı ve 1 saatte +5ºC’ye düşürülerek 2 saat süre ile ekilibrasyona bırakıldı. Ekilibre edilen sperma 0.25 ml’lik payetlere çekilerek iki farklı soğutma hızında (yavaş: +5°C’tan -20°C’a 0.5°C/dak ve hızlı: +5°C’tan -20°C’a 5°C/dak) donduruldu. Her iki grup -20°C’tan -120°C’a 25°C/dak hızla donduruldu. Eritme sonrası (37°C/30 dak) aşamada; akrozom bozukluğu Pisum sativum agglutinin fluorescein conjugate ( FITC PSA) ile kromatin yapısı Acridin Orange (AO) ile ve apoptozis ise terminal deoxynucleotidyl transferase-mediated dUTP nick-end labeling tekniği (TUNEL) kullanarak belirlendi. Motilite muayeneleri ısıtma tablalı faz kontrast mikroskop ile gerçekleştirildi. Eritme sonrası motilite, akrozom bozukluğu, AO ve TUNEL değerleri sırasıyla yavaş dondurma için %42.8±8.8, %31.6±12.9, %2.9±2.4 ve 2.8±1.6 hızlı dondurma için ise %36.5±9.9, %24.7±11.1, %3.3±2.2 ve %6.3±3.4 olarak saptandı. Spermanın eritme sonrası bulgularının değerlendirilmesinde akrozom bozukluğu ve sperma kromatin yapısı (AO) yönünden iki soğutma hızı arasında fark bulunmazken, eritme sonrası motilitede gruplar arasında istatistiksel fark bulundu (P<0.05). Sonuç olarak, koç spermasında yavaş soğutma prosedürü ile dondurma, eritme sonrası motiliteyi iyileştirirken, akrozom ve kromatin bütünlüğünü ve apoptotik indeksi soğutma oranları arasında önemli bir fark göstermemiştir.

Anahtar Kelime: spermatozoa dondurulmuş semen koç programlanmış hücre ölümü donma koyun semen özellikleri kromatin çözülme erkek döl verimi akrozom reaksiyonu

Konular: Veterinerlik

Koç spermasında dondurma hızının akrozom ve kromatin bütünlüğü üzerine etkisi

Öz:

The objective of the present study was to investigate the effect of different freezing rates on post-thaw sperm motility, acrosome defect, and sperm chromatin structure and apoptotic activity in ram semen. Collected semen was diluted at 1:5 (semen/extender) with Bioxel® (IMV technologies France) at 30ºC and then cooled to 5ºC within 1h. Cooled semen was subjected to the equilibration for 2 hours. Equilibrated semen was frozen in 0.25 ml straw at two different cooling rates (slow: 0.5°C/min from 5 to -20°C and fast: 5°C/min from 5 to -20°C). Both groups were frozen from -20 to -120°C at 25°C/min and stored in liquid nitrogen until use. Post-thaw (37°C/30 min) sperm motility, defected acrosome (Pisum sativum agglutinin fluorescein conjugate, FITC PSA), sperm chromatin structure determined by Acridin Orange (AO) and apoptotic activity using terminal deoxynucleotidyl transferasemediated dUTP nick-end labeling (TUNEL) were evaluated. Post-thaw sperm motility, acrosome defect, AO and TUNEL for slow frozen semen were 42.8±8.8%, 31.6±12.9%, 2.9±2.4% and 2.8±1.6%, and for fast frozen semen were 36.5±9.9%, 24.7±11.1%, 3.3±2.2% and 6.3±3.4%, respectively. Post-thaw semen analyses showed that there was no significant difference between two freezing curves in terms of acrosome defect, sperm chromatin damage (AO). However, a significant difference was found for postthaw semen motility between two groups (P<0.05). In conclusion, while the slow freezing procedure improved post-thaw sperm motility, acrosome and chromatin integrities and apoptotic index in ram spermatozoa did not show any significant difference between freezing rates.

Anahtar Kelime: apoptosis freezing sheep semen characters chromatin thawing male fertility acrosome reaction spermatozoa frozen semen rams

Konular: Veterinerlik

Belge Türü: Makale Makale Türü: Araştırma Makalesi Erişim Türü: Erişime Açık

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APA	NUR Z, ZIK B, ÜSTÜNER B, TÜTÜNCÜ Ş, SAĞIRKAYA H, OZGUDEN G, GÜNAY Ü, DOĞAN İ (2011). Effect of freezing rate on acrosome and chromatin integrity in ram semen. , 267 - 272.
Chicago	NUR ZEKARIYA,ZIK Berrin,ÜSTÜNER BURCU,TÜTÜNCÜ ŞERİFE,SAĞIRKAYA Hakan,OZGUDEN G. Cansel,GÜNAY Ülgen,DOĞAN İbrahim Effect of freezing rate on acrosome and chromatin integrity in ram semen. (2011): 267 - 272.
MLA	NUR ZEKARIYA,ZIK Berrin,ÜSTÜNER BURCU,TÜTÜNCÜ ŞERİFE,SAĞIRKAYA Hakan,OZGUDEN G. Cansel,GÜNAY Ülgen,DOĞAN İbrahim Effect of freezing rate on acrosome and chromatin integrity in ram semen. , 2011, ss.267 - 272.
AMA	NUR Z,ZIK B,ÜSTÜNER B,TÜTÜNCÜ Ş,SAĞIRKAYA H,OZGUDEN G,GÜNAY Ü,DOĞAN İ Effect of freezing rate on acrosome and chromatin integrity in ram semen. . 2011; 267 - 272.
Vancouver	NUR Z,ZIK B,ÜSTÜNER B,TÜTÜNCÜ Ş,SAĞIRKAYA H,OZGUDEN G,GÜNAY Ü,DOĞAN İ Effect of freezing rate on acrosome and chromatin integrity in ram semen. . 2011; 267 - 272.
IEEE	NUR Z,ZIK B,ÜSTÜNER B,TÜTÜNCÜ Ş,SAĞIRKAYA H,OZGUDEN G,GÜNAY Ü,DOĞAN İ "Effect of freezing rate on acrosome and chromatin integrity in ram semen." , ss.267 - 272, 2011.
ISNAD	NUR, ZEKARIYA vd. "Effect of freezing rate on acrosome and chromatin integrity in ram semen". (2011), 267-272.

APA	NUR Z, ZIK B, ÜSTÜNER B, TÜTÜNCÜ Ş, SAĞIRKAYA H, OZGUDEN G, GÜNAY Ü, DOĞAN İ (2011). Effect of freezing rate on acrosome and chromatin integrity in ram semen. Ankara Üniversitesi Veteriner Fakültesi Dergisi, 58(4), 267 - 272.
Chicago	NUR ZEKARIYA,ZIK Berrin,ÜSTÜNER BURCU,TÜTÜNCÜ ŞERİFE,SAĞIRKAYA Hakan,OZGUDEN G. Cansel,GÜNAY Ülgen,DOĞAN İbrahim Effect of freezing rate on acrosome and chromatin integrity in ram semen. Ankara Üniversitesi Veteriner Fakültesi Dergisi 58, no.4 (2011): 267 - 272.
MLA	NUR ZEKARIYA,ZIK Berrin,ÜSTÜNER BURCU,TÜTÜNCÜ ŞERİFE,SAĞIRKAYA Hakan,OZGUDEN G. Cansel,GÜNAY Ülgen,DOĞAN İbrahim Effect of freezing rate on acrosome and chromatin integrity in ram semen. Ankara Üniversitesi Veteriner Fakültesi Dergisi, vol.58, no.4, 2011, ss.267 - 272.
AMA	NUR Z,ZIK B,ÜSTÜNER B,TÜTÜNCÜ Ş,SAĞIRKAYA H,OZGUDEN G,GÜNAY Ü,DOĞAN İ Effect of freezing rate on acrosome and chromatin integrity in ram semen. Ankara Üniversitesi Veteriner Fakültesi Dergisi. 2011; 58(4): 267 - 272.
Vancouver	NUR Z,ZIK B,ÜSTÜNER B,TÜTÜNCÜ Ş,SAĞIRKAYA H,OZGUDEN G,GÜNAY Ü,DOĞAN İ Effect of freezing rate on acrosome and chromatin integrity in ram semen. Ankara Üniversitesi Veteriner Fakültesi Dergisi. 2011; 58(4): 267 - 272.
IEEE	NUR Z,ZIK B,ÜSTÜNER B,TÜTÜNCÜ Ş,SAĞIRKAYA H,OZGUDEN G,GÜNAY Ü,DOĞAN İ "Effect of freezing rate on acrosome and chromatin integrity in ram semen." Ankara Üniversitesi Veteriner Fakültesi Dergisi, 58, ss.267 - 272, 2011.
ISNAD	NUR, ZEKARIYA vd. "Effect of freezing rate on acrosome and chromatin integrity in ram semen". Ankara Üniversitesi Veteriner Fakültesi Dergisi 58/4 (2011), 267-272.