%0 Journal Article
%T Aging of stallion spermatozoa stored in vitro is delayed at 22°C using a 67 mm glucose-10 mm pyruvate-based media.
%A Becerro-Rey L
%A Martín-Cano FE
%A Ferrusola CO
%A Rodríguez-Martínez H
%A Gaitskell-Phillips G
%A da Silva-Álvarez E
%A Silva-Rodríguez A
%A Gil MC
%A Peña FJ
%J Andrology
%V 12
%N 5
%D 2024 Jul 1
%M 38041502
%F 4.456
%R 10.1111/andr.13565
%X <B>BACKGROUND: </B>Most commerce of equine seminal doses is carried out using commercial extenders under refrigeration at 5°C.<BR><B>OBJECTIVE: </B>To 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.<BR><B>METHODS: </B>Stallion 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.<BR><B>RESULTS: </B>After 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.<BR><B>CONCLUSIONS: </B>Aliquots 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+.