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


NUR Z., ZIK B., ÜSTÜNER B., Tutuncu S., SAĞIRKAYA H., ÖZGÜDEN AKKOÇ C. G., ...Daha Fazla

ANKARA UNIVERSITESI VETERINER FAKULTESI DERGISI, cilt.58, sa.4, ss.267-272, 2011 (SCI-Expanded) identifier identifier

  • Yayın Türü: Makale / Tam Makale
  • Cilt numarası: 58 Sayı: 4
  • Basım Tarihi: 2011
  • Doi Numarası: 10.1501/vetfak_000000248
  • Dergi Adı: ANKARA UNIVERSITESI VETERINER FAKULTESI DERGISI
  • Derginin Tarandığı İndeksler: Science Citation Index Expanded (SCI-EXPANDED), Scopus, TR DİZİN (ULAKBİM)
  • Sayfa Sayıları: ss.267-272
  • Anahtar Kelimeler: Apoptosis, freezing rate, ram semen, CRYOPROTECTIVE AGENTS, MEMBRANE INTEGRITY, SPERMATOZOA FROZEN, BULL SPERM, FERTILITY, EXTENDER, GLYCEROL, CRYOPRESERVATION, INSEMINATION, OSMOLALITY
  • Bursa Uludağ Üniversitesi Adresli: Evet

Özet

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 (R) (IMV technologies France) at 30 degrees C and then cooled to 5 degrees C 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 degrees C/min from 5 to -20 degrees C and fast: 5 degrees C/min from 5 to -20 degrees C). Both groups were frozen from -20 to -120 degrees C at 25 degrees C/min and stored in liquid nitrogen until use. Post-thaw (37 degrees 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 transferase-mediated 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 post-thaw 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.