Lactate-induced spontaneous acrosomal exocytosis as a method to study acrosome function in stallion sperm

Hernández‐Avilés, Camilo; Ramírez‐Agámez, Luisa; Varner, D.D.; Love, Charles C.

doi:10.1016/j.theriogenology.2023.07.024

Cited by 3 publications

(1 citation statement)

References 86 publications

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“…The results warrant further follow-up research on the metabolism of the spermatozoa impacting current sperm technologies, as is also demonstrated in recent reports on the mechanism of sperm capacitation in horses. 59,60…”

Section: Discussionmentioning

confidence: 99%

Aging of stallion spermatozoa stored in vitro is delayed at 22°C using a 67 mm glucose–10 mm pyruvate‐based media

Becerro‐Rey,

Martín‐Cano,

Ferrusola

et al. 2023

Andrology

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BackgroundMost commerce of equine seminal doses is carried out using commercial extenders under refrigeration at 5°C.ObjectivesTo determine if 10 mm pyruvate in a 67 mm glucose extender and storage at 22°C could be the basis of an alternative storage method to cooling to 5°C.Material and methodsStallion ejaculates were extendedin: INRA96 (67 mm glucose, non‐pyruvate control), modified Tyrode's (67 mm glucose–10 mm pyruvate), supplemented with 0, 10, 50, and 100 μM itaconate. As itaconate was vehiculated in DMSO, a control vehicle was also included. Sperm motility, viability, mitochondrial membrane potential, and production of reactive oxygen species were measured after collection and again after 48 and 96 h of storage at 22°C. To disclose molecular metabolic changes, spermatozoa were incubated up to 3 h in modified Tyrode's 67 mm glucose–10 mm pyruvate and modified Tyrode's 67 mm glucose, and metabolic analysis conducted.ResultsAfter 96 h of storage aliquots stored in the control, INRA96 had a very poor total motility of 5.6% ± 2.3%, while in the 67 mm glucose–10 mm pyruvate/10 μm itaconate extender, total motility was 34.7% ± 3.8% (p = 0.0066). After 96 h, viability was better in most pyruvate‐based media, and the mitochondrial membrane potential in spermatozoa extended in INRA96 was relatively lower (p < 0.0001). Metabolomics revealed that in the spermatozoa incubated in the high pyruvate media, there was an increase in the relative amounts of NAD+, pyruvate, lactate, and ATP.Discussion and conclusionsAliquots stored in a 67 mm glucose–10 mm pyruvate‐based medium supplemented with 10 μM itaconate, maintained a 35% total motility after 96 h of storage at 22°C, which is considered the minimum acceptable motility for commercialization. Improvements may be related to the conversion of pyruvate to lactate and regeneration of NAD+.

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Section: Discussionmentioning

confidence: 99%

Aging of stallion spermatozoa stored in vitro is delayed at 22°C using a 67 mm glucose–10 mm pyruvate‐based media

Becerro‐Rey,

Martín‐Cano,

Ferrusola

et al. 2023

Andrology

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Proteomic analysis of sperm from fertile stallions and subfertile stallions due to impaired acrosomal exocytosis

Hernández-Avilés,

Ramírez-Agámez,

Weintraub

et al. 2024

Sci Rep

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Thoroughbred stallions that carry a double-homozygous genotype A/A-A/A for SNPs rs397316122 and rs69101140 in exon 5 of the FKBP6 gene (chr13; EquCab3.0) are uniquely subfertile due to impaired acrosomal exocytosis (IAE). In this study, the sperm proteome in frozen/thawed semen from subfertile Thoroughbred stallions was studied and compared to that of frozen/thawed sperm from fertile Thoroughbred stallions. A total of 2,220 proteins was identified, of which 140 proteins were found to be differentially abundant in sperm from the subfertile stallions compared to that of fertile stallions (83 less and 57 more abundant). Proteins of differential abundance in sperm from the subfertile stallions were mainly overrepresented in the “metabolism” and the “metabolism of lipids” pathways. One of these proteins, arylsulfatase F (ARSF), was studied by immunofluorescence. A lower proportion of sperm displaying ARSF signal at the acrosome region was observed in sperm from subfertile Thoroughbred stallions. In addition, heterologous zona pellucida binding assays revealed that sperm from subfertile Thoroughbred stallions bound at a lower proportion to zonae pellucidae than sperm from fertile Thoroughbred stallions. In conclusion, a group of differential abundance proteins, including some of acrosome origin, were identified in sperm from subfertile stallions with acrosome dysfunction.

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Proteomic analysis of sperm from fertile stallions and subfertile stallions due to impaired acrosomal exocytosis

Hernández-Avilés,

Ramírez-Agámez,

Weintraub

et al. 2024

Preprint

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Thoroughbred stallions that carry a double-homozygous genotype A/A-A/A for SNPs rs397316122 and rs69101140 in exon 5 of the FKBP6 gene (chr13; EquCab3.0) are uniquely subfertile due to impaired acrosomal exocytosis (IAE). In this study, the sperm proteome in frozen/thawed semen from subfertile Thoroughbred stallions was studied and compared to that of frozen/thawed sperm from fertile Thoroughbred stallions. A total of 2,220 proteins was identified, of which 140 proteins were found to be differentially abundant in sperm from the subfertile stallions when compared to that of fertile stallions (83 less and 57 more abundant). Proteins of differential abundance in sperm from the subfertile stallions were mostly overrepresented in the “metabolism” and the “metabolism of lipids” pathways. One of these proteins, arylsulfatase F (ARSF), was studied by immunofluorescence. A lower proportion of sperm displaying ARSF signal at the acrosome region was observed in sperm from subfertile Thoroughbred stallions. In addition, heterologous zona pellucida binding assays were performed and revealed sperm from subfertile Thoroughbred stallions bound at a lower proportion to zonae pellucidae than sperm from fertile Thoroughbred stallions. In conclusion, a group of proteins of differential abundance, including some of acrosome origin, were identified in sperm from subfertile stallions with acrosome dysfunction.

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Lactate-induced spontaneous acrosomal exocytosis as a method to study acrosome function in stallion sperm

Cited by 3 publications

References 86 publications

Aging of stallion spermatozoa stored in vitro is delayed at 22°C using a 67 mm glucose–10 mm pyruvate‐based media

Aging of stallion spermatozoa stored in vitro is delayed at 22°C using a 67 mm glucose–10 mm pyruvate‐based media

Proteomic analysis of sperm from fertile stallions and subfertile stallions due to impaired acrosomal exocytosis

Proteomic analysis of sperm from fertile stallions and subfertile stallions due to impaired acrosomal exocytosis

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