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. 2022 Nov 21;19(4):e20220056.
doi: 10.1590/1984-3143-AR2022-0056. eCollection 2022.

Action of swim-up and caffeine on equine frozen sperm

Natália de Castro Alves  1 , Soraia de Araújo Diniz  1 , Rodrigo Novaes Viegas  1 , Ana Luiza Arigoni  1 , Marina Morra Freitas  1 , Ângela Quintão Lana  1 , Monique de Albuquerque Lagares  1
Affiliations

Action of swim-up and caffeine on equine frozen sperm

Natália de Castro Alves et al. Anim Reprod. .

Abstract

Cryopreservation of equine semen is crucial to semen commercialization. However, it reduces sperm motility and longevity. Thus, sperm selection methods and addition of motility-activating substances to sperm, such as caffeine, may improve sperm quality of equine frozen semen. The objective of the current work was to evaluate the effects of caffeine on recovery and quality parameters of frozen-thawed sperm subjected to swim-up selection to be used in intracytoplasmic sperm injection (ICSI) in assisted reproductive techniques. Stallion semen were frozen and after thawing different caffeine concentrations were added to the samples performing four treatments control (no caffeine), 3, 5, and 7.5 mM caffeine. Sperm kinematic and motility were assessed by computer-assisted sperm analysis (CASA). Then, the four treated samples were submitted to the swim-up sperm selection, and the number of recovered sperm and morphology were evaluated at four times 20, 40, 60, and 80 min. The swim-up increased the recovery proportion of normal morphology sperm without (80.1±1%) or with caffeine addition (3mM: 81.2±1%, 5mM: 79.9±1% and 7.5 mM 78.9±1%) compared to the thawed semen (70±2%). However, the addition of 5 mM caffeine induced an increase in sperm motility (38.9±2.8 vs. 32.6±3.4%, P<0.05), and sperm recovery after swim-up (7.9x106 vs. 3.4x106 sperm/ml, P<0.05) compared to the control. The addition of 5 mM caffeine to frozen-thawed equine semen before swim-up selection improved sperm motility and increased the sperm recovery rate while not decreasing the percentage of morphologically normal sperm. Thus, caffeine addition to frozen-thawed equine semen before swim-up selection has potential clinical application in improving sperm quality for use in ICSI.

Keywords: cryopreservation; freezing; sperm selection; spermatozoa; stallion.

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Conflict of interest statement

Conflicts of interest: The authors have no conflict of interest to declare.

Figures

Figure 1
Figure 1. Scheme of the pathways of caffeine action on sperm energy production inhibiting phosphodiesterase enzyme activity on cAMP degradation (a) and binding to the adenosine receptor stimulating adenylyl cyclase converting ATP into cAMP (b). Both pathways cause an increase of intracellular concentration of cAMP activating protein kinase (PKA), increasing cytochrome c oxidase (COX) activity and oxidative phosphorylation.
Figure 2
Figure 2. Scheme of swim-up.
See this image and copyright information in PMC

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