Apoptosis in fresh and cryopreserved buffalo sperm

Khan, D.R.; Ahmad, Nasim; Anzar, M.; Channa, Aijaz Ali

doi:10.1016/j.theriogenology.2008.09.056

Cited by 35 publications

(28 citation statements)

References 14 publications

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“…Translocation of PS from the inner to the outer leaflet takes place in early apoptosis [15] as well as during sperm capacitation [16] and after incubation with bicarbonate [17]. Damaged membranes owing to temperature effects have been shown to bring about impaired sperm motility [18], but whether in vivo heat exposure induces changes in the membrane phospholipid organization that results in subsequent reduced sperm viability appears to be unknown.…”

Section: Introductionmentioning

confidence: 99%

Whole-body heat exposure induces membrane changes in spermatozoa from the cauda epididymidis of laboratory mice

Wechalekar¹,

Setchell²,

Peirce³

et al. 2010

Asian J Androl

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This study was carried out to determine if exposure to hot environmental temperatures had a direct, detrimental effect on sperm quality. For this the effect of whole-body heat exposure on epididymal spermatozoa of laboratory mice was investigated. C57BL/6 mice (n = 7) were housed in a microclimate chamber at 37ºC-38ºC for 8 h per day for three consecutive days, while control mice (n = 7) were kept at 23ºC-24ºC. Cauda epididymal spermatozoa were obtained 16 h after the last heat treatment. The results showed that sperm numbers were similar in the two groups (P = 0.23), but after heat treatment, a significant reduction in the percentage of motile sperm was present (P < 0.0001). Membrane changes of the spermatozoa were investigated by staining with phycoerythrin (PE)-conjugated Annexin V, which detects exteriorization of phosphotidylserine from the inner to the outer leaflet of the sperm plasma membrane, and 7-aminoactinomycin D (7-AAD), which binds to the sperm nucleus when the plasma membrane is damaged. The percentage of spermatozoa showing positive staining with Annexin V-PE or 7-AAD or both, was significantly higher (P < 0.05) in heat-exposed mice compared with controls. These results show that whole-body heat exposure to 37ºC-38ºC induces membrane changes in the epididymal spermatozoa of mice, which may lead to apoptosis.

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Section: Introductionmentioning

confidence: 99%

Whole-body heat exposure induces membrane changes in spermatozoa from the cauda epididymidis of laboratory mice

Wechalekar¹,

Setchell²,

Peirce³

et al. 2010

Asian J Androl

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“…La motilidad es el indicador más sensible del daño celular debido a la congelación, incluso más que la integridad de la membrana plasmática (Yu et al), efecto que también ha sido observado en espermatozoides de carnero (Del Valle et al, 2010) y búfalo (Khan et al, 2009). Los resultados obtenidos en el presente estudio, concuerdan con lo sugerido por Yu et al, quien obtuvo porcentajes de MOP más bajos que los porcentajes con membrana plasmática íntegra, atribuyendo esta diferencia a cambios en el transporte activo y a la permeabilidad en la membrana plasmática de la región de la cola.…”

Section: Y Benson Et Alunclassified

Criopreservación de Espermatozoides Caninos a - 80C

2013

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RESUMEN. La congelación y almacenamiento en nitrógeno líquido (N 2 L) es la técnica más utilizada para preservar el semen de canino y de otras especies domésticas durante largos períodos de tiempo. Este estudio fue diseñado para validar el uso del ultracongelador de -80ºC para congelar y almacenar semen canino en pajuelas. Tres protocolos fueron ensayados (n=10): Control: semen congelado y almacenado en N 2 L; Exp. 1: semen congelado y almacenado -80ºC, y Exp.2: semen congelado a -80ºC y almacenado en N 2 L. Postdescongelación se evaluó por citometría de flujo la viabilidad e integridad de membrana plasmática (SYBR-14/PI), potencial de membrana mitocondrial (∆Ψm; JC-1), integridad de la membrana del acrosoma (PSA/FITC-PI), translocación de fosfatidilserina (Annexin-V-FITC/PI) y fragmentación del ADN (TUNEL). No se observaron diferencias significativas (P<0,05) entre los grupos Control, Exp.1 y Exp.2 respecto a: integridad de membrana plasmática intacta (42,2 ± 5,3; 35,57 ± 10,3 y 40,76 ± 12,1; respectivamente), ∆Ψm normal (54,7 ± 20,6; 44,4 ± 15,8 y 43,4 ± 15,3 respectivamente), membrana acrosomal intacta (42,9 ± 11,1; 53,2 ± 15,8 y 48,7 ± 20,1 respectivamente) y ADN fragmentado (0,87 ± 0,41; 0,75 ± 0,16; 0,79 ± 0,27 respectivamente). Sin embargo, el promedio de la motilidad progresiva postdescongelación de los grupos E1 y E2 (37,0 ± 4,4%; 36,8 ± 4,1% respectivamente) demostraron diferencias (P<0,05) respecto al grupo Control (55,9 ± 8,7%). Los resultados obtenidos en el presente estudio demostraron que la utilización del ultracongelador de -80ºC para congelar y almacenar espermatozoides caninos tiene un uso potencial en medicina veterinaria como alternativa a la utilización del N 2 L. PALABRAS CLAVE: Canino; Espermatozoide; Congelación; Ultracongelador de -80ºC. INTRODUCCIÓNEn caninos, la congelación y conservación de espermatozoides por periodos prolongados de tiempo se realiza en nitrógeno líquido (N 2 L) (Alamo et al., 2005). Además, han sido utilizadas técnicas alternativas para el procedimiento de congelación como el hielo seco y ultra-congeladores programables para conservar muestras de semen, sin embargo el almacenamiento es en N 2 L (Olar et al., 1989;Hay et al., 1997;Thomas et al., 1993).Ha sido determinada la función espermática en ultracongeladores de -152ºC utilizando parámetros como la motilidad, reacción del acrosoma y morfoanomalías (Ála-mo et al.;Batista et al., 2006). Sin embargo, no existen antecedentes de la utilización del ultracongelador de -80ºC, para congelar semen canino en pajuelas convencionales por periodos cortos como alternativa a la técnica convencional de congelación en N 2 L. En este estudio se evalúa el efecto de la temperatura de congelación y almacenamiento a -80ºC, en comparación al método convencional de congelación y almacenamiento en N 2 L, sobre parámetros esenciales de la función del espermatozoide de canino, tales como motilidad progresiva, funcionalidad de membranas (plasmática, acrosomal y mitocondrial) y fragmentación del ADN. MATERIAL Y MÉTODOAnimales. Se utilizaron...

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“…This assertion is supported when it is observed that in the same treatment the D-Me rises to almost 60%, accompanied by a significant correlation with F-DNA, suggesting a common origin. In fact, authors such as Khan et al (2009), found that the apoptosis increases significantly after cryopreservation, and that this variable is positively correlated with the membrane damages, both at a significant rate after cryopreservation.…”

Section: Dna Fragmentation During Cryopreservation and Its Relationshmentioning

confidence: 99%

“…Many of the factors that contend the integrity of the membrane during cryopreservation can simultaneously affect the DNA, such as the external and internal formation of ice crystals in the cell, ultimately causing mechanical cellular damage (Miskolczi et al, 2005); or events like apoptosis (Khan et al, 2009), which can trigger DNA fragmentation with the help of the caspases and, by means of secondary effects, disintegrate the membranes (Enari, 1998;Goulet et al, 1998), events induced by chemical, physical, or pharmalogical factors (Thompson,1995), as those present during the cryopreservation process, such as, the external and internal cryoprotectants such as glucose and Dimethylsulfoxide (DMSO).…”

Section: Introductionmentioning

confidence: 99%

DNA fragmentation and membrane damage of bocachico Prochilodus magdalenae (Ostariophysi: Prochilodontidae) sperm following cryopreservation with dimethylsulfoxide and glucose

2012

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The endangered bocachico Prochilodus magdalenae is a native freshwater fish of Colombia, the most captured species locally and one of the most important species for ex-situ conservation (germplasm banks). The aim of this study was to examine the effect of three concentrations of Dimethylsulfoxide (DMSO) (5%, 10%, 15%) and three of glucose (305, 333, 361 mM) in the extender on spermatic DNA fragmentation (F-DNA) (by Halomax®, Chromatin dispersion) and membrane damage (D-Me) (by eosin-nigrosin staining). After assessment of sperm quality by computer analysis of motility, one part of semen from males was diluted separately with three parts of extender and filled into 0.5 ml straws. Freezing was carried out in liquid nitrogen vapor dry shipper for 30 minutes and thawed at 60°C for 8 seconds in a water bath and evaluated for the percentage of cells found with F-DNA and D-Me. The results demonstrated that cryopreservation causes greater F-DNA (13.62 ± 1.6% to 28.91 ± 3.25) and D-Me (24.27 ± 1.1% to 58.33 ± 2.81%) when compared with pre-freezing semen (PFS) (6.71 ± 1.54% and 2.34 ± 0.5%, respectively for F-DNA and D-Me). A significant interaction was found between DMSO and glucose concentration in this experiment. Use of extender: 10% DMSO + 305 mM glucose + 12% chicken egg yolk and, 10% DMSO + 333 mM glucose + 12% chicken egg yolk, allow for lower F-DNA and D-Me during cryopreservation of bocachico semen. A high correlation between F-DNA and D-Me was found (r = 0.771).O curimba Prochilodus magdalenae, é uma espécie nativa de água doce da Colômbia ameaçada de extinção, sendo a mais capturada localmente e uma das mais importantes para a conservação ex-situ (bancos de germoplasma). O objetivo deste estudo foi avaliar o efeito de três concentrações de dimetilsulfóxido (DMSO) (5%, 10%, 15%) e três de glicose (305, 333, 361 mM) no diluente sobre a fragmentação do ADN espermático (F-DNA) (através de Halomax®, dispersão da cromatina) e danos em membrana (D-Me) (através da coloração eosina-nigrosina). Depois de avaliar a qualidade espermática por análise computacional da mobilidade, uma parte do sêmen dos machos foi diluída separadamente com três partes do diluente e colocado dentro de canudos de 0.5 ml. O congelamento foi feito em tanque de vapores secos de nitrogênio líquido por 30 minutos e descongelado a 60°C por 8 segundos em banho de água e avaliada a percentagem de células com F-DNA e D-Me. Os resultados demonstraram que a criopreservação causa grande F-DNA (13.62 ± 1.6% até 28.91 ± 3.25) e D-Me (24.27 ± 1.1% até 58.33 ± 2.81%) quando comparada com o sêmen pré-congelamento (PFS) (6.71 ± 1.54% e 2.34 ± 0.5%, respectivamente para F-DNA e D-Me). Uma interação significativa foi encontrada entre a concentração de DMSO e glicose neste experimento. O uso dos diluentes 10% DMSO + 305 mM glicose + 12% gema de ovo de galinha e 10% DMSO + 333 mM glicose + 12% gema de ovo de galinha permitem obter a menor F-DNA e D-Me durante a criopreservação do sêmen de curimba. Uma alta correlação entre F-DNA e D-Me foi encontrada (r = 0.771).

show abstract

Apoptosis in fresh and cryopreserved buffalo sperm

Cited by 35 publications

References 14 publications

Whole-body heat exposure induces membrane changes in spermatozoa from the cauda epididymidis of laboratory mice

Whole-body heat exposure induces membrane changes in spermatozoa from the cauda epididymidis of laboratory mice

Criopreservación de Espermatozoides Caninos a - 80C

DNA fragmentation and membrane damage of bocachico Prochilodus magdalenae (Ostariophysi: Prochilodontidae) sperm following cryopreservation with dimethylsulfoxide and glucose

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