Changes in the phospholipid composition and phospholipid asymmetry of ram sperm plasma membranes after cryopreservation

Hinkovska‐Galcheva, Vania; Petkova, Diana; Koumanov, Kamen

doi:10.1016/0011-2240(89)90034-5

Cited by 47 publications

(18 citation statements)

References 24 publications

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“…This goes with the previously observed plasma membrane changes: the spermatozoal cell membrane displays certain reordering of their lipid components, leading to a loss of stability of the lipid bilayer (Schiller et al 2000). In turn, sperm membrane integrity becomes impaired with translocation of phosphatidylserine from the inner to the outer leaflet of the sperm plasma membrane (Glander and Schaller 1999;Hinkovska-Galcheva et al 1989). This translocation is considered one of the early signs of terminal phase of apoptosis (Martin et al 1995).…”

Section: Discussionmentioning

confidence: 59%

Magnetic-activated Cell Sorting before Cryopreservation Preserves Mitochondrial Integrity in Human Spermatozoa

Grunewald

Paasch

Said

et al. 2006

Cell Tissue Banking

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Superparamagnetic annexin-V conjugated microbeads are able to eliminate spermatozoa with externalized phosphatidylserine, a membrane feature of apoptotic cells as well as spermatozoa with deteriorated plasma membrane. Our objective was to evaluate the effects of annexin-V Magnetic-Activated Cell Sorting (MACS) in cryopreservation-thawing protocols and on integrity of sperm mitochondrial transmembrane potential and mitochondrial integrity survival rate (MSR). Mature spermatozoa of 10 healthy donors were prepared by density gradient centrifugation and divided into 2 aliquots afterwards. The first one was subjected to annexin-V MACS followed by cryopreservation and thawing, while the second was cryopreserved-thawed without MACS to serve as control. Annexin-negative sperm separated by MACS showed significantly higher levels of intact mitochondria following cryopreservation-thawing (45.4+/-8.6%) compared to sperm that were not separated (15.8+/-4.6%, p<0.01). Separating a distinctive population of non-apoptotic spermatozoa with intact membranes may optimize cryopreservation-thawing outcome. MACS using annexin-V microbeads enhances the percentage of spermatozoa with intact transmembrane mitochondrial potential and mitochondrial integrity survival rates following cryopreservation.

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Section: Discussionmentioning

confidence: 59%

Magnetic-activated Cell Sorting before Cryopreservation Preserves Mitochondrial Integrity in Human Spermatozoa

Grunewald

Paasch

Said

et al. 2006

Cell Tissue Banking

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“…It is possible that the nonregulated cryocapacitation arises following the osmotic and mechanical stresses to the plasma membrane during freezing and thawing [47]. The plasma membrane is the primary site of sperm cryoinjury [28,48] and undergoes various modifications during cryopreservation, including changes to fluidity [49], phospholipid profiles, and phospholipid distribution [50,51]. The increased permeability to calcium observed in cryopreserved sperm [44,52] might simply be a direct consequence of the mechanical damage to the sperm plasma membrane.…”

Section: Glucose Does Not Inhibit Cryocapacitationmentioning

confidence: 98%

A Differential Mechanism Is Involved During Heparin- and Cryopreservation-Induced Capacitation of Bovine Spermatozoa

Cormier

Bailey

2003

Biology of Reproduction

151

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After ejaculation, mammalian spermatozoa must undergo capacitation to fertilize. Capacitation of bovine spermatozoa occurs in vitro in medium supplemented with heparin. Semen cryopreservation is an important tool for assisted reproduction, although the fertility of frozen-thawed spermatozoa is reduced, possibly due to precocious capacitation-like changes that are known to occur. Our purpose was to clarify the mechanisms involved in bull sperm cryocapacitation induced by cryopreservation. Our general hypothesis is that the signaling pathways that lead to capacitation are triggered by the cryopreservation procedure. Ejaculated bovine semen was divided into two aliquots and diluted in extender; one was then kept fresh, whereas the second was cryopreserved. Western blots of extracted sperm proteins with anti-phosphotyrosine antibody showed that capacitation, induced by either heparin in fresh sperm or cryopreservation (cryocapacitation), is associated with a differential profile of phosphotyrosine-containing proteins. Immunolocalization of phosphotyrosine-containing proteins in the fresh and cryopreserved spermatozoa showed that, after thawing, cryocapacitated sperm displayed labeling over the acrosomal region, whereas for fresh sperm, this labeling appeared after 5-h incubation with heparin. The chlortetracycline assay and the ability of the sperm to undergo the lysophosphatidylcholine-induced acrosome reaction were used to confirm that a subpopulation of cryopreserved sperm is capacitated at thawing, irrespective of heparin inclusion. Since glucose is known to inhibit heparin-induced capacitation, the semen extender was modified to include glucose as a means of inhibiting cryocapacitation; however, cryocapacitation was not prevented according to the chlortetracycline assay and profile of phosphotyrosine-containing sperm proteins.

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“…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. Cryopreservation impacts lipid composition and the organization of the sperm plasma membrane [1] . In addition, sudden temperature changes, such as cold shocks, ice formation and dissolution during the freezing-…”

Section: öZetunclassified

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.

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Changes in the phospholipid composition and phospholipid asymmetry of ram sperm plasma membranes after cryopreservation

Cited by 47 publications

References 24 publications

Magnetic-activated Cell Sorting before Cryopreservation Preserves Mitochondrial Integrity in Human Spermatozoa

Magnetic-activated Cell Sorting before Cryopreservation Preserves Mitochondrial Integrity in Human Spermatozoa

A Differential Mechanism Is Involved During Heparin- and Cryopreservation-Induced Capacitation of Bovine Spermatozoa

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

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