Effects of cryopreservation procedures on sperm membranes

Parks, John E.; Graham, J. K.

doi:10.1016/0093-691x(92)90231-f

Cited by 323 publications

(201 citation statements)

References 45 publications

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“…Therefore, they are less likely either to survive long enough in the female reproductive tract or to maintain competency in fertilization, as compared to fresh-extended semen [14].…”

Section: Discussionmentioning

confidence: 99%

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Fixed-time AI pregnancy rate following insemination with frozen-thawed or fresh-extended semen in progesterone supplemented CO-Synch protocol in beef cows

et al. 2009

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Fixed-time AI pregnancy rate following insemination with frozen-thawed or fresh-extended semen in progesterone supplemented CO-Synch protocol in beef cows SummaryThe objective of this study was to compare fixed-time AI pregnancy rate in Angus crossbred beef cows inseminated with frozen-thawed or fresh-extended semen. Two ejaculates from each of two Angus bulls were collected by artificial vagina and pooled for each bull. The pooled semen from each bull was divided into two aliquots; Aliquot 1 was extended using Caprogen ® (LIC, Hamilton, New Zealand) to a concentration of 3 × 10 6 sperm/straw and Aliquot 2 was extended using egg-yolk-glycerol extender to a concentration of 20 × 10 6 sperm/straw.Semen extended with Caprogen ® was maintained at ambient temperature and semen extended with egg-yolk-glycerol extender was frozen and maintained at GnRH im and were inseminated at a fixed-time on Day 10, 66 h after CIDR removal. Timed-AI pregnancy rates were influenced by season (P < 0.05), cows detected in estrus prior to and at AI (P < 0.001), and dam age (P < 0.01).Pregnancy rates were not affected by semen type (Fresh = 51.5% vs. Frozen = 50.4%; P = 0.66) and there were no significant interactions of semen type by estrus expression, semen type by sire, or semen type by season (P > 0.1). In conclusion, commercial beef cows inseminated with fresh-extended semen (3 × 10 6 sperm/straw) yielded comparable pregnancy rates to conventional frozenthawed semen in a progesterone supplemented, CO-Synch fixed-time AI synchronization protocol and may provide an alternative to frozen semen for more efficient utilization of superior genetics.

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“…Therefore, they are less likely either to survive long enough in the female reproductive tract or to maintain competency in fertilization, as compared to fresh-extended semen [14].…”

Section: Discussionmentioning

confidence: 99%

“…The addition of catalase at temperatures over 5 °C and the reduction of egg-yolk to 5% as a substrate of the enzyme significantly reduced H2O2 concentrations [14]. Caprogen ® extender is commonly used in NZ to inseminate cows, using fresh semen at lower concentrations.…”

Section: Introductionmentioning

confidence: 99%

Fixed-time AI pregnancy rate following insemination with frozen-thawed or fresh-extended semen in progesterone supplemented CO-Synch protocol in beef cows

et al. 2009

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“…The plasma membrane is the primary site of injury to cryopreserved sperm and the principal damage occurs during freezing and thawing (Hammerstedt et al 1990;Parks and Graham 1992). As sperm are frozen, ice crystals form in the extracellular medium, thereby increasing the osmolality of the unfrozen water.…”

Section: The Cryopreservation Processmentioning

confidence: 99%

“…Certainly the phospholipids in egg yolk are considered to support the sperm plasma membrane during the stress of processing [reviewed in Parks and Graham (1992)], . Ejaculated sperm were extended with egg yolk-Trisglycerol and half were cryoperserved then incubated for 5 h in capacitating medium with 0 (lanes 1, 3 and 5) or 15 µg mL -1 heparin (lanes 2, 4 and 6).…”

Section: How Can Sperm Be Protected?mentioning

confidence: 99%

Semen cryopreservation: Successes and persistent problems in farm species

Bailey

Morrier²,

Cormier³

2003

Can. J. Anim. Sci.

121

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Bailey, J., Morrier, A. and Cormier. N. 2003. Semen cryopreservation: Successes and persistent problems in farm species. Can. J. Anim. Sci. 83: 393-401. Artificial insemination has arguably been the most important practice contributing to the advancement of animal production. The numerous advantages of artificial insemination are augmented when the semen is cryopreserved and conserved for long periods. Unfortunately, the utility of cryopreserved semen is limited because for most mammals, even cattle, a considerable proportion of sperm loose their fertility during freezing-thawing. In many species, this loss of fertility is substantial, rendering cryopreserved semen impractical for routine use. Our goals are to characterize the nature of the sublethal sperm damage caused by cryopreservation, and to develop a method to improve the functional fertility of thawed semen. This review describes our theory that sperm are prematurely activated by cryopreservation ("cryo-capacitation") and discusses strategies for preventing this damage.Key words: Sperm, artificial insemination, capacitation, fertility Bailey, J., Morrier, A. et Cormier, N. 2003. La cryoconservation de la semence : les succès et les problèmes persistants chez les espèces de la ferme. Can. J. Anim. Sci. 83: 393-401. L'insémination artificielle a grandement contribué à l'avancement des productions animales. Les nombreux avantages qu'offre cette technique sont augmentés lorsque la semence est cryoconservée et entreposée pour de longues périodes. Malheureusement, l'utilisation de la semence cryoconservée est limitée puisque chez la majorité des mammifères, incluant la vache, une large proportion des spermatozoïdes perdra leur motilité pendant le processus de congéla-tion-décongélation. Chez plusieurs espèces, cette perte de fertilité est suffisamment substantielle pour rendre impraticable l'utilisation routinière de la semence cryoconservée. Les objectifs de notre laboratoire portent sur la caractérisation des dommages sous létaux subis par les spermatozoïdes lors de la cryoconservation et le développement de méthodes permettant d'améliorer la fertilité des spermatozoïdes décongelés. Cette revue décrit notre théorie selon laquelle les spermatozoïdes seraient prématurément activés par la cryoconservation ("cryo-capacitation"). Différentes stratégies permettant de prévenir ce dommage y sont également abordées.

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“…The plasma membrane is the primary site of injury in cryopreserved sperm (Parks & Graham 1992); however, the increase in the proteins concentration of in the media can augment the stability of the sperm membrane, reducing the chances of alterations induced by refrigeration (DeLeeuw et al 1990). Concerning this, the addition of egg yolk to the extender may have improved the resistance of the plasma membrane in some armadillos' sperm during the freezing--thawing step.…”

Section: Discussionmentioning

confidence: 99%

Identification of ultrastructural and functional damages in sperm from six-banded armadillos (Euphractus sexcinctus) due to cryopreservation

et al. 2016

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RESUMO.-[Identificação de danos ultraestruturais e funcionais em espermatozoides de tatus-peba (Euphractus sexcinctus) oriundos da criopreservação.] O objetivo deste estudo foi criopreservar o sêmen de tatus-peba (Euphractus sexcinctus) em diluente Tris-gema e glicerol, e determinar os danos causados pelo processo de congelação-descongelação, utilizando marcadores fluorescentes e análise ultraestrutural. As amostras de sêmen (n=11) coletadas de 4 tatus-peba adultos por eletroejaculação foram criopreservadas em diluente Tris acrescido de 20% de gema de ovo e 3% de glicerol, em curva rápida de congelação. A análise clássica das amostras foi realizada após a diluição, refrigeração e descongelação, seguida por análise de fluorescência, utilizando uma combinação de sondas flu- The aim of the study was to cryopreserve the semen of six-banded armadillos (Euphractus sexcinctus) in Tris-yolk and glycerol diluent, and to determine the damage caused by the freezing-thawing process, using fluorescent markers and ultrastructural analysis. Semen samples (n=11) collected from 4 adult six-banded armadillos by electroejaculation were cryopreserved in Tris diluent plus 20% egg yolk and 3% glycerol, in a fast freezing curve. Classical analysis of samples was performed after dilution, refrigeration and thawing, followed by fluorescence analysis, using a combination of fluorescent probes to assess membrane integrity (propidium iodide -PI and Hoechst -H342), and mitochondrial activity (CMXRos -Mito Tracker Red ® ). We also used the ultrastructural analysis to verify possible morphological alterations caused by cryoinjuries. When compared with fresh samples, we verified a significant decline in all the armadillos' semen parameters after thawing, in which only 6.1% motile sperm were found. However, the percentage of sperm which remained with viable (13%) and functional (24.7%) membranes after thawing suggests that some cells could be live but immotile. Analysis using fluorescent markers revealed that the mitochondria of armadillos' sperm is highly sensible to the freezing protocol and the findings through ultrastructure analysis proved this statement. Additionally, the images obtained by transmission electron microscopy revealed that frozen-thawed sperm presented damaged plasma membrane, nuclear modifications as changes in chromatin and acrossomal changes relative to sperm capacitation. In conclusion, this study is the first attempt to cryopreserve the semen of an armadillo species, and to help us to identify critical points on the freezing-thawing procedure in order to improve the protocol. Além disso, as imagens obtidas por microscopia eletrônica de transmissão revelaram que espermatozoides congelados-descongelados apresentaram membranas plasmáticas danificadas, modificações nucleares como alterações na cromatina, e alterações acrossomais relativas à capacitação espermática. Em conclusão, este estudo é a primeira tentativa de criopreservação de sêmen em uma espécie de tatu, e nos auxiliou a identificar pontos críticos no processo de ...

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Effects of cryopreservation procedures on sperm membranes

Cited by 323 publications

References 45 publications

Fixed-time AI pregnancy rate following insemination with frozen-thawed or fresh-extended semen in progesterone supplemented CO-Synch protocol in beef cows

Fixed-time AI pregnancy rate following insemination with frozen-thawed or fresh-extended semen in progesterone supplemented CO-Synch protocol in beef cows

Semen cryopreservation: Successes and persistent problems in farm species

Identification of ultrastructural and functional damages in sperm from six-banded armadillos (Euphractus sexcinctus) due to cryopreservation

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