Optimizing conditions for treating goat semen with cholesterol-loaded cyclodextrins prior to freezing to improve cryosurvival

Konyali, C.; Tomás, C.; Blanch, E.; Gómez, Ernesto A.; Graham, J. K.; Mocé, E.

doi:10.1016/j.cryobiol.2013.06.001

Cited by 45 publications

(30 citation statements)

References 53 publications

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“…The enrichment of CLC in sperm membrane increased the membrane fluidity during cooling and cryopreservation and, decreased upon thawing which enhance the cryosurvival rate [34]. Through this method, the sperm cryosurvival rate of large variety of species including bull [33], ram [24], buck [15], buffalo [35], bison [13], mouse [26], elephant [14], rabbit [28], donkey [30], fish [46] and camel [9] has dramatically been increased. However, some species' sperm like swine [45], equine [27], rooster [44] and feline [32] did not show any improvement in motility, viability and acrosome integrity except membrane integrity.…”

Section: Introductionmentioning

confidence: 99%

Protective effect of cholesterol-loaded cyclodextrin pretreatment against hydrogen peroxide induced oxidative damage in ram sperm

et al. 2015

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Section: Introductionmentioning

confidence: 99%

Protective effect of cholesterol-loaded cyclodextrin pretreatment against hydrogen peroxide induced oxidative damage in ram sperm

et al. 2015

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“…Ejaculates were diluted with Tris extender and centrifuged, as previously described, to eliminate seminal plasma. After washing, each ejaculate was split into two treatment aliquots, one served as a control (non-CLC treated) and the other one was treated with 1 mg of CLC/120 × 10 6 sperm in accordance with the protocol described by Konyali et al (2013). Samples were incubated for 15 min at 22°C, after which each sample was diluted to 200 × 10 6 cells/ml with EYT.…”

Section: Direct Freezing Of Bull and Goat Spermmentioning

confidence: 99%

“…Ejaculates were processed according to the protocol described by Konyali et al (2013). After collection, semen was washed twice by centrifugation to remove the seminal plasma.…”

Section: Clc Preparationmentioning

confidence: 99%

Effect of cholesterol-loaded cyclodextrins on bull and goat sperm processed with fast or slow cryopreservation protocols

Mocé

Tomás

Blanch

et al. 2014

Animal

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Cholesterol-loaded cyclodextrins (CLC) added to the sperm before cryopreservation enhance sperm quality after freeze-thawing in several cold shock-sensitive species, including cattle and goats. However, all studies conducted to date have used conventional protocols, in which sperm are cooled slowly to 5°C before freezing. As cholesterol plays a significant role in sperm cold shock resistance, it is possible that CLC-treated sperm can withstand cooling damage when the sperm are not cooled slowly to 5°C before freezing. In this study, we determined whether CLC-treated goat (1 mg CLC/120 × 10 6 sperm) and bull (2 mg CLC/120 × 10 6 sperm) sperm quality, after thawing, was different for sperm frozen using conventional protocols (including a slow cooling phase to 5ºC) and protocols in which the sperm were frozen from room temperature, without cooling the sperm slowly to 5°C before freezing. CLC-treated sperm exhibited higher percentages of plasma membrane-intact sperm than control sperm when cryopreserved using conventional protocols. In addition, CLC treatment enhanced both sperm motility and plasma membrane integrity when sperm were frozen directly from room temperature. However, this treatment did not fully prevent the damage of the sperm after cooling rapidly and subsequent freezing, as the sperm quality was lower than that presented by the samples frozen using the conventional protocol. The results are promising, but studies to optimize the protocols for freezing sperm directly from room temperature need to be conducted, as well as studies to determine how cryopreserving sperm in this manner affects other sperm functions.Keywords: cooling rate, semen, cattle, buck ImplicationsFreezing protocols for the semen of most of the species include a slow cooling phase to 5ºC. This phase requires from refrigeration units and comprises at least 85% of the total time cryopreserving sperm. In this study, we investigate whether treating bull and goat sperm with cholesterolloaded cyclodextrins allows eliminating this cooling phase. Our results show that this treatment partially prevents the damage to the sperm when directly frozen from room temperature and with this technique the sperm cryopreservation protocols are simplified, which would allow the processing of more ejaculates per day or freezing ejaculates collected in the field.

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“…Durante años se han desarrollado diferentes técnicas para la obtención de muestras, así como metodologías para su almacenamiento en condiciones no fisiológicas para asegurar la disponibilidad futura y diversidad del material genético procedente de diferentes especies domésticas (Blackburn, 2012). En caprino se han probado diferentes protocolos de criopreservación de semen, sin embargo, tras el proceso de dilución y estrés osmótico, así como los procesos de enfriamiento y congelación-descongelación, la fertilidad del semen se ve fuertemente afectada (Konyali et al, 2013). La disminución de la fertilidad durante el proceso de criopreservación es especialmente relevante en caprino debido a los efectos negativos que tiene la reacción enzimática entre la fosfolipasa del plasma seminal (producida en las glándulas bulbouretrales) y la lecitina que contiene la yema de huevo del diluyente de congelación, que resulta perjudicial debido a la formación de lisolecitina que es tóxica para los espermatozoides (Leboeuf et al, 2000).…”

Section: Introduccionunclassified

Dinámicas espermáticas subpoblacionales durante el proceso de criopreservación en eyaculados de caprino (Capra aegagrus hircus)

Barbas¹,

Horta²,

García-Herreros³

2017

SPERMOVA

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RESUMENEl objetivo del presente estudio fue determinar la dinámica espermática subpoblacional mediante la evaluación objetiva de parámetros cinéticos funcionales durante la criopreservación del semen de caprino. Se utilizaron 40 eyaculados procedentes de 8 machos en los que se evaluaron los parámetros de volumen, concentración, viabilidad, integridad acrosómica y motilidad espermática objetiva mediante sistema computarizado de análisis seminal (CASA). Se evaluaron un total de 8 parámetros cinéticos (VCL, VSL, VAP, LIN, STR, ALH, BCF y WOB) en cada etapa de manipulación de la muestra espermática durante el periodo de criopreservación [1º: semen fresco (F); 2º: 1er lavado/centrifugación (1LC); 3º: 2º lavado/centrifugación (2LC); 4º: enfriamiento o "cooling" a 4ºC (CL) y 5º: pos-descongelación a 37ºC (PD)]. Los resultados derivados de los parámetros cinéticos espermáticos se analizaron mediante análisis de componentes principales (PCA) y análisis multivariante de conglomerados para identificar la relación entre las subpoblaciones espermáticas obtenidas y los parámetros funcionales de motilidad en cada etapa. Se detectaron cuatro subpoblaciones espermáticas en la 2ª, 3ª y 5ª etapa (Sbp1LC1-Sbp1LC4, Sbp2LC1-Sbp2LC4 y SbpPD1-SbpPD4) y tres en la 1ª (SbpF1-SbpF3) y 4ª etapa (SbpCL1-SbpCL3). Según los parámetros de velocidad obtenidos en cada subpoblación diferenciada se observaron células rápidas, lentas, activas, pasivas, progresivas y no progresivas. Por otro lado, tomando en cuenta los parámetros de linealidad se observaron trayectorias espermáticas curvilíneas, rectilíneas, parabólicas y erráticas. Se observó variabilidad en las subpoblaciónes detectadas a lo largo de las etapas de manipulación evaluadas habiendo múltiples diferencias significativas entre los parámetros cinéticos obtenidos intra e inter-subpoblacionales (p<0,001) así como en la frecuencia de distribución de las subpoblaciones obtenidas entre etapas (p<0,05). En conclusión, el presente estudio confirmó la existencia de variabilidad y heterogeneidad en los parámetros cinéticos espermáticos en el eyaculado caprino a lo largo del proceso de criopreservación. La relación entre la estructura subpoblacional espermática y las diferentes etapas de manipulación fue estadísticamente significativa, especialmente en las etapas de enfriamiento o "cooling" y posdescongelación debido a que éstas alteraron de forma drástica los parámetros cinéticos espermáticos.Palabras clave: Subpoblaciones espermáticas, criopreservación, semen, caprino. ABSTRACTThe objective of the present research was to determine specific sperm subpopulational dynamics in different processing steps during cryopreservation process by using objective functional sperm kinematic descriptors in goat ejaculates. Fresh ejaculates (n=40) collected from eight bucks were analised for volume, concentration, sperm viability, acrosome integrity, and sperm motility using computer-assisted sperm analysis (CASA) system. Eight sperm kinematic descriptors (VCL, VSL, VAP, LIN, STR, BCF, ALH, and WOB) were assesse...

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Optimizing conditions for treating goat semen with cholesterol-loaded cyclodextrins prior to freezing to improve cryosurvival

Cited by 45 publications

References 53 publications

Protective effect of cholesterol-loaded cyclodextrin pretreatment against hydrogen peroxide induced oxidative damage in ram sperm

Protective effect of cholesterol-loaded cyclodextrin pretreatment against hydrogen peroxide induced oxidative damage in ram sperm

Effect of cholesterol-loaded cyclodextrins on bull and goat sperm processed with fast or slow cryopreservation protocols

Dinámicas espermáticas subpoblacionales durante el proceso de criopreservación en eyaculados de caprino (Capra aegagrus hircus)

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