RT info:eu-repo/semantics/article
T1 Washing increases the susceptibility to exogenous oxidative stress in red deer spermatozoa
A1 Domínguez Rebolledo, Álvaro Efrén
A1 Fernández Santos, María Rocío
A1 García Álvarez, Olga
A1 Maroto Morales, Alejandro
A1 Garde López-Brea, Julián
A1 Martínez Pastor, Felipe
A2 Biologia Celular
K1 Veterinaria
K1 Antioxidants
K1 Centrifugation
K1 Oxidative stress
K1 Red deer
K1 Washing
AB The effects of routine sperm work are often overlooked. We assessed the effect of washing cryopreserved epididymal spermatozoa from red deer (Cervus elaphus hispanicus, Helzheimer 1909). After thawing, epididymal samples (four stags) were diluted in TALP-HEPES. A split was left untouched, another was centrifuged (300 × g, 5 min) and resuspended, and a third was centrifuged and the supernatant substituted by fresh TALP-HEPES (washing). Each split was supplemented either with nothing, 1 mM of the antioxidant Trolox, 100 μM of the oxidant Fe (with ascorbate), or both. The 3 × 4 treatments were incubated at 37°C and assessed each hour up to 3 h for motility (computer-aided sperm assessment) and viability/apoptosis plus mitochondrial status (YO-PRO-1, propidium iodide, Mitotracker Deep Red; flow cytometry). DNA damage at 4 h was assessed using the terminal deoxynucleotidyl transferase–mediated dUTP nick end-labeling assay. Centrifugation alone affected neither sperm quality nor DNA, and the oxidant had no effect in control or centrifuged samples. Washed samples were not different than control, but oxidant decreased motility, mitochondrial status and viability, and altered the motility subpopulation pattern, being partially suppressed by Trolox. Spermatozoa with damaged DNA dramatically increased in the washed-oxidized sample (from 22.30 ± 3.52% to 67.94 ± 5.07%), but not when antioxidant was present. Although samples from different males behaved similarly, male-to-male variability was detected regarding susceptibility to oxidative damage after washing. We concluded that, although red deer thawed spermatozoa seemed resilient to centrifugation, the vulnerability to oxidative stress after washing makes it advisable to supplement manipulation media with antioxidants, especially taking into account male-to-male variability.
PB Elsevier
YR 2019
FD 2019-04-22
LK http://hdl.handle.net/10612/10529
UL http://hdl.handle.net/10612/10529
NO Theriogenology, 2009, vol. 72, n. 8
NO P. 1073-1084
DS BULERIA. Repositorio Institucional de la Universidad de León
RD 27-abr-2024