Dissecting the molecular damage to stallion spermatozoa: The way to improve current cryopreservation protocols?

Peña, Fernando J.; Macías‐García, Beatriz; Samper, J.C.; Aparicio, I.M.; Tapia, José A.; Ortega-Ferrusola, Cristina

doi:10.1016/j.theriogenology.2011.06.023

Cited by 121 publications

(97 citation statements)

References 118 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…Efforts have been made to improve the composition of extenders used for both refrigeration and cryopreservation, by adding antioxidants, cryoprotectants, and other similar reagents (Ball and Vo, 2001;Ball et al, 2001;Guthrie et al, 2002;Aboagla and Terada, 2003;Pena et al, 2011;Lagares et al, 2012;Morillo-Rodriguez et al, 2012;Karimfar et al, 2015). More recently, the study of molecular mechanisms controlling cell survival or cell death has emerged as a successful approach to improve semen conservation and to extend the lifespan of the sperm in vitro (Watson, 2000;Ortega Ferrusola et al, 2010;Cordova et al, 2014).…”

Section: Introductionmentioning

confidence: 99%

The autophagy-related protein LC3 is processed in stallion spermatozoa during short-and long-term storage and the related stressful conditions

Aparicio

Muñoz

Salido

et al. 2016

Animal

Self Cite

View full text Add to dashboard Cite

Use of cooled and frozen semen is becoming increasingly prevalent in the equine industry. However, these procedures cause harmful effects in the sperm cell resulting in reduced cell lifespan and fertility rates. Apoptosis and necrosis-related events are increased during semen cryopreservation. However, a third type of cell death, named autophagy, has not been studied during equine semen storage. Light chain (LC)3 protein is a key component of the autophagy pathway. Under autophagy activation, LC3-I is lipidated and converted to LC3-II. The ratio of LC3-II/LC3-I is widely used as a marker of autophagy activation. The main objective of this study was to investigate whether LC3 is processed during cooling, freezing and the stressful conditions associated with these technologies. A secondary objective was to determine if LC3 processing can be modulated and if that may improve the quality of cryopreserved semen. LC3 processing was studied by Western blot with a specific antibody that recognized both LC3-I and LC3-II. Viability was assessed by flow cytometry. Modulation of LC3-I to LC3-II was studied with known autophagy activators (STF-62247 and rapamycin) or inhibitors (chloroquine and 3-MA) used in somatic cells. The results showed that conversion of LC3-I to LC3-II increased significantly during cooling at 4°C, freezing/thawing and each of the stressful conditions tested (UV radiation, oxidative stress, osmotic stress and changes in temperature). STF-62247 and rapamycin increased the LC3-II/LC3-I ratio and decreased the viability of equine sperm, whereas chloroquine and 3-MA inhibited LC3 processing and maintained the percentage of viable cells after 2 h of incubation at 37°C. Finally, refrigeration at 4°C for 96 h and freezing at −196°C in the presence of chloroquine and 3-MA resulted in higher percentages of viable cells. In conclusion, results showed that an 'autophagy-like' mechanism may be involved in the regulation of sperm viability during equine semen cryopreservation. Modulation of autophagy during these reproductive technologies may result in an improvement of semen quality and therefore in higher fertility rates.Keywords: cryopreservation, refrigeration, viability, autophagy, sperm ImplicationsReproductive technologies with cooled and frozen semen have unquestionable advantages in animal breeding; however, semen preservation results in important injuries in the sperm cells, compromising their viability and hence the pregnancy rates. In the last decade, the scientific community has increased its efforts to improve current cryopreservation protocols. The results of the current study demonstrate that LC3, a protein involved in autophagy, is activated under stressful conditions during cryopreservation, and plays a role in regulating viability. Therefore, this study is of interest not only from a scientific point of view, but also for the improvement of semen quality, which could result in high fertility and finally increased economic gain.

show abstract

Section: Introductionmentioning

confidence: 99%

The autophagy-related protein LC3 is processed in stallion spermatozoa during short-and long-term storage and the related stressful conditions

Aparicio

Muñoz

Salido

et al. 2016

Animal

Self Cite

View full text Add to dashboard Cite

show abstract

“…It is documented that the semen cooling process causes several changes in mammal spermatozoa known as a whole as 'cold shock' (Watson, 2000). It is particularly focused on changes in sperm membranes (Peña et al, 2011); however, this negative effect alters cellular metabolism and organelles, decreases motility and induces irregularities in the sperm motility pattern (Sieme et al, 2008). As a consequence, sperm quality decreases during the cooling process and so do pregnancy rates (Varner et al, 1989).…”

Section: Introductionmentioning

confidence: 99%

Effect of single layer centrifugation using Androcoll-E-Large on the sperm quality parameters of cooled-stored donkey semen doses

Ortiz

Dorado

Ramirez

et al. 2014

Animal

View full text Add to dashboard Cite

The aim of this study was to determine the effect of single layer centrifugation (SLC) using Androcoll-E-Large on donkey sperm quality parameters after 24 h of cool-storage. Ejaculates were collected from Andalusian donkeys and then cooled at 5°C. SLC was carried out after 24 h of cool-storage using Androcoll-E-Large. In the first experiment, all sperm parameters assessed (total and progressive sperm motility, viability, sperm morphology and sperm kinematics VCL, VSL, VAP, LIN, STR, WOB, ALH and BCF) were statistically compared between semen samples processed or not with Androcoll-E-Large. Significant differences ( P < 0.05) were found between SLC-selected and unselected semen samples for all parameters assessed, obtaining better results after SLC. In the second experiment, semen samples were classified in two groups according to their sperm progressive motility (PM) before SLC. Then, the increments obtained in semen quality parameters after SLC were compared between groups. No significant differences were found between groups, indicating that SLC improved the sperm quality parameters of entire set of semen samples processed with independence to their original PM. In conclusion, SLC with Androcoll-E-Large can be used in donkeys, increasing the sperm quality of cooled-stored donkey semen doses after 24 h of cool storage.

show abstract

“…Amides have proven to be a mostly beneficial CPA, having been shown to decrease damaging results compared to those obtained when glycerol is solely used as the CPO [28]. With stallions being sorted into different freezing classes, poor freezers have required substantial work.…”

Section: Amidesmentioning

confidence: 99%

Cryoprotectants & Cryopreservation of Equine Semen: A Review of Industry Cryoprotectants and the Effects of Cryopreservation on Equine Semen Membranes

Prien¹

2016

JDVAR

View full text Add to dashboard Cite

show abstract

Dissecting the molecular damage to stallion spermatozoa: The way to improve current cryopreservation protocols?

Cited by 121 publications

References 118 publications

The autophagy-related protein LC3 is processed in stallion spermatozoa during short-and long-term storage and the related stressful conditions

The autophagy-related protein LC3 is processed in stallion spermatozoa during short-and long-term storage and the related stressful conditions

Effect of single layer centrifugation using Androcoll-E-Large on the sperm quality parameters of cooled-stored donkey semen doses

Cryoprotectants & Cryopreservation of Equine Semen: A Review of Industry Cryoprotectants and the Effects of Cryopreservation on Equine Semen Membranes

Contact Info

Product

Resources

About