Effects of different cryoprotective agents on ram sperm morphology and DNAintegrity

Nur, Zekariya; Zık, Berrin; Üstüner, Burcu; Sağırkaya, Hakan; Ozguden, C.

doi:10.1016/j.theriogenology.2009.12.007

Cited by 87 publications

(90 citation statements)

References 41 publications

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“…Extender composition assists with the stabilization of cells during the freezing and thawing processes [2,14] . Extenders containing soybean lecithin could be an alternative to the conventional extenders that include egg yolk [5,18] .…”

Section: Discussionmentioning

confidence: 99%

“…Semen was collected by electrically stimulated ejaculation (Ruakura Ram Probe Plastic Products, Hamilton, New Zealand) [2,15] . Ram semen was collected five times in every other day.…”

Section: Semen Collection Evaluation and Dilutionmentioning

confidence: 99%

“…Equilibrated semen was placed into 0.25 ml straws and frozen at 3°C/min from +5°C to -8°C and at 25°C/min from -8°C to -120°C in liquid nitrogen vapor using the Nicool Plus PC freezing machine (Air Liquide, Marne-la-Vallée Cedex 3, France) [2] . The straws were then plunged into liquid nitrogen at -196°C where they were stored for at least one month.…”

Section: Semen Freezing and Thawingmentioning

confidence: 99%

“… İletişim (Correspondence)  +90 224 2941245  bbaspinar@uludag.edu.tr thawing process, affect the integrity and function of the acrosome, nucleus, mitochondria, axonema and plasma membrane [2][3][4] .…”

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

Effect of Egg Yolk and Soybean Lecithin on Tris-Based Extender in Post-Thaw Ram Semen Quality and in vitro Fertility

Üstüner¹,

Alçay²,

Nur³

et al. 2014

Kafkas Univ Vet Fak Derg

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The aim of the current study was to evaluate the effect of egg yolk and different soybean lecithin concentrations on the efficiency of ram semen cryopreservation and to test the fertilizing ability of frozen-thawed ram semen. Ejaculates with a thick consistency, rapid wave motion (3-5), and >70% initial motility were pooled. Pooled semen were then divided into four groups and diluted at 1/5 (semen/extender) with 1%, 3%, 6% lecithin (L1, L3 and L6) or 20% egg yolk (EY20) using the two-step dilution method. As expected, the results of the current study showed that both motility and the rates of defective acrosomes in sperm were negatively affected by the cryopreservation procedure (P<0.001). The motility values of at 5°C and post-thawed semen in the EY20 group were significantly higher than those in the L1, L3 and L6 groups (P<0.05). There were no differences in motility rates among the lecithin groups at the dilution, cooling, equilibration or post thawing stages (P>0.05). The results of in vitro fertilization, as assessed by the rate of blastocyst formation, were more successful in the EY20 group than those noted in different lecithin groups. In conclusion, freezing ram semen with an extender containing egg yolk could yield better post-thaw sperm parameters and embryonic development compared to lecithin containing extenders. Keywords: Cryopreservation, Lecithin, Egg yolk, Ram semen, IVF Tris-Bazlı Sulandırıcılarda Yumurta Sarısı ve Soya Lesitininin Eritme Sonrası Koç Spermasının Kalitesi ve in vitro Fertilite Üzerine Etkisi ÖzetBu çalışmanın amacı, yumurta sarısı ve farklı lesitin konsantrasyonlarının koç spermasının dondurulabilirliği üzerine etkisini değerlendirmek ve dondurup çözdürülen koç spermasının fertilizasyon yeteneğini tespit etmektir. Kitle hareketi (3-5) ve >%70 motiliteye sahip ejakülatlar birleştirildi (pooling). Pooling yapılan sperma dört gruba bölündü ve %1, %3, %6 (L1, L3 ve L6) lesitin veya %20 yumurta sarısı (EY20) içeren sulandırıcılar ile 1/5 (sperma/sulandırıcı) oranında iki aşamalı sulandırma yöntemi kullanılarak sulandırıldı. Bu çalışmanın sonucunda; motilite ve akrozomal bozukluk oranlarının dondurma prosedüründen olumsuz yönde etkilendiği tespit edildi (P<0.001). EY20 grubunun 5°C'de ve eritme sonrası motilite değerleri L1, L3 ve L6 gruplarına göre yüksek bulundu (P<0.05). Sulandırma, soğutma, ekilibrasyon ve eritme sonrası aşamalarda motilite oranları bakımından lesitin grupları arasında farklılık saptanmadı (P>0.05). Blastosist oranları bakımından değerlendirilen in vitro fertilizasyon sonuçlarına göre, EY20 grubunun farklı lesitin gruplarından daha başarılı olduğu tespit edildi. Sonuç olarak, yumurta sarısı içeren sulandırıcı ile dondurulan koç spermasının lesitin gruplarına göre eritme sonrası spermatolojik özellikler ve embriyonik gelişim bakımından daha üstün olduğu tespit edildi.

show abstract

Section: Discussionmentioning

confidence: 99%

“…Semen was collected by electrically stimulated ejaculation (Ruakura Ram Probe Plastic Products, Hamilton, New Zealand) [2,15] . Ram semen was collected five times in every other day.…”

Section: Semen Collection Evaluation and Dilutionmentioning

confidence: 99%

Section: Semen Freezing and Thawingmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Effect of Egg Yolk and Soybean Lecithin on Tris-Based Extender in Post-Thaw Ram Semen Quality and in vitro Fertility

Üstüner¹,

Alçay²,

Nur³

et al. 2014

Kafkas Univ Vet Fak Derg

View full text Add to dashboard Cite

show abstract

“…Windsor and White (1995) reported severe mitochondrial damage of ram spermatozoa following freezing and thawing and reasoned that the associated attenuation of respiratory metabolism may compromise their cervical passage. Recently, Nur et al (2010) reported the effects of a range of cryoprotective agents (glycerol, propanediol, sucrose and trehalose) on DNA integrity. All of them induced apoptosis and DNA damage.…”

Section: Recent Studies In Semen Freezingmentioning

confidence: 99%

Traversing the ovine cervix – a challenge for cryopreserved semen and creative science

Robinson

McKelvey²,

King

et al. 2011

Animal

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This review brings together research findings on cervical relaxation in the ewe and its pharmacological stimulation for enhancement of the penetration needed for transcervical insemination and embryo transfer. On the basis that the success of artificial insemination is the percentage of ewes lambing, a review is made of recent research aimed at understanding and minimising the sub-lethal effects of freezing and thawing on the viability of spermatozoa, their membrane integrity and their ability to migrate through cervical mucus, as these characteristics have a major influence on fertility, particularly when semen is deposited, artificially, in the os cervix. Milestones of achievement are given for transcervical intrauterine insemination, embryo recovery and transfer and the birth of lambs of pre-determined sex, firstly following intracytoplasmic sperm injection, then laparoscopic intrauterine insemination using highly diluted flow-cytometrically sorted fresh semen and subsequently by os cervix insemination using sexed semen that had been frozen and thawed. Diversity of research endeavour (applied, cellular, molecular), research discipline (anatomy, histology, immunology, endocrinology) and research focus (cell, tissue, organ, whole animal) is embraced within the review as each has significant contributions to make in advancing recent scientific findings from the laboratory into robust on-farm transcervical insemination and embryo transfer techniques.Keywords: ovine cervix, semen cryopreservation, artificial insemination, embryo recovery and transfer ImplicationsIn bringing together the results of recent research on the cryopreservation of ram semen and the relaxation of the ewe cervix, this review provides a basis for identifying opportunities for improving the efficiency of cervical insemination and transcervical embryo recovery and transfer in sheep genetic improvement programmes.

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The effects of supplementation of vitamin D to the egg‐yolk extender on cryopreservation of ram semen

Aktar,

Toker,

Koca

et al. 2024

Veterinary Medicine & Sci

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ObjectivesThis study aimed to examine the effects of supplementation of vitamin D to the egg‐yolk extender on characteristics of frozen–thawed ram semen.MethodsSemen samples obtained from adult rams were pooled and divided into five equal volumes. It was reconstituted with extenders containing different concentrations of vitamin D: 0 (control), 12.5 (VITD 12.5), 25 (VITD 25), 50 (VITD 50), and 100 ng/mL (VITD 100), and then they were frozen. Sperm motility parameters, plasma membrane functional integrity, acrosomal integrity, DNA fragmentation, and mitochondrial membrane potential of the groups were evaluated after sperm thawing.ResultsTotal motility and progressive motility were higher in VITD 50 than in all other groups (p < 0.05). Higher sperm straightness, linearity, and wooble were higher in VITD 50 than in the control group (p < 0.05). A similar pattern of VITD 50 was observed for plasma membrane integrity and mitochondrial membrane potential (p > 0.05).ConclusionsIn the study, it was observed that adding vitamin D to the extender had a beneficial effect on ram spermatological parameters. In addition, it was concluded that the use of the 50 ng/mL vitamin D in the extender provided more effective protection than the other doses.

show abstract

Effects of different cryoprotective agents on ram sperm morphology and DNAintegrity

Cited by 87 publications

References 41 publications

Effect of Egg Yolk and Soybean Lecithin on Tris-Based Extender in Post-Thaw Ram Semen Quality and in vitro Fertility

Effect of Egg Yolk and Soybean Lecithin on Tris-Based Extender in Post-Thaw Ram Semen Quality and in vitro Fertility

Traversing the ovine cervix – a challenge for cryopreserved semen and creative science

The effects of supplementation of vitamin D to the egg‐yolk extender on cryopreservation of ram semen

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