New assays for detection and localization of endogenous lipid peroxidation products in living boar sperm after BTS dilution or after freeze–thawing

Brouwers, Jos F.; Silva, Patrícia F.N.; Gadella, B.M.

doi:10.1016/j.theriogenology.2004.09.046

Cited by 106 publications

(61 citation statements)

References 56 publications

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“…This pattern is very similar to bovine, porcine, stallion and human spermatozoa [Brouwers and Gadella 2003;Brouwers et al 2005;Neild et al 2005;]. As pointed out by Brouwers and Gadella [2003], the absence of LPO in the sperm head could be due to the presence of a potent antioxidant system able to defend the paternal DNA.…”

Section: Discussionsupporting

confidence: 73%

Lipid Peroxidation in Human Spermatozoa from Men with Genitourinary Infections

Cosci

Moretti

Collodel

2008

Systems Biology in Reproductive Medicine

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A fluorescent assay was used to reveal the presence of lipid peroxidation (LPO) in spermatozoa from individuals with genitourinary infections (GI). The incidence of LPO was compared with sperm pathologies (apoptosis, immaturity, necrosis) evaluated by transmission electron microscopy (TEM) and motility. A C11-BODIPY 581/591 probe was used to detect and localize LPO in sperm from 34 individuals. The sperm morphological characteristics were studied by TEM and the results were mathematically assessed. Using the LPO percentage we divided the patients into four groups. Group 1 included ten patients with GI with almost normal progressive motility, a very low percentage of LPO and sperm pathology values that were not far from the standard ranges. Group 2 had eleven infected patients showing reduced motility, LPO of 10 to 20% and sperm pathologies that were just out of normal range. Group 3 included 6 infected patients with progressive motility r22%, low levels of LPO with a higher percentage of apoptosis and necrosis. Group 4 had 7 patients showing normal semen parameters, without GI, LPO ranged from 0% to 1% and a normal incidence of sperm pathologies. In all groups, LPO was inversely correlated with sperm motility and directly correlated with low semen quality. However, the LPO percentage was low in Group 3, in which sperm necrosis, concomitant with GI, was the predominant pathology. C11-BODIPY 581/591 is a useful labeling method for quantifying and localizing LPO in spermatozoa from patients with GI. The detection of LPO could be questionable in cases of sperm necrosis because in this pathology the plasma membrane is often disrupted and the lipidic probe is unable to intercalate within the phospholipidic bilayer.

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Section: Discussionsupporting

confidence: 73%

Lipid Peroxidation in Human Spermatozoa from Men with Genitourinary Infections

Cosci

Moretti

Collodel

2008

Systems Biology in Reproductive Medicine

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“…The plasma membrane defects and the loss of important intracellular ingredients (such as calcium ions, adenine nucleotides, antioxidants, and enzymes) induce metabolic disruption that may affect sperm motility and vitality. Coolinginduced reductions in sperm function may also be due to oxidative damage from the harmful formation of reactive oxygen species (ROS) and membrane lipid peroxidation (Brouwers et al, 2005;Aitken, 2006;Großfeld et al, 2008;Awda et al, 2009). In addition to the plasma membrane, mitochondria are the sperm structures most sensitive to the stress induced by cooling and freezing (Flores et al, 2009;Peña et al, 2009).…”

Section: Introductionmentioning

confidence: 99%

Storage temperature of boar semen and its relationship to changes in sperm plasma membrane integrity, mitochondrial membrane potential, and oxidoreductive capability

Gączarzewicz¹,

Udała²,

Piasecka³

et al. 2015

Turk J Biol

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The aim of this study was to analyze changes in sperm plasma membrane integrity, mitochondrial activity, and extracellular environment during storage of boar semen at 5 °C, 16 °C, and 25 °C for 10 days. Progressive sperm motility, plasma membrane integrity (SYBR-14/PI-test and HOS test), aspartate aminotransferase (AAT), and mitochondrial activity (JC-1-test and NADH-dependent NBT assay), as well as pH and osmolality were assessed in nine ejaculates of Polish Landrace boars. The plasma membrane integrity of the semen stored at 5 °C was similar to that of the semen stored at 16 °C and 25 °C; however, an increase in AAT activity of the semen stored at 5 °C revealed sperm membrane disorders as early as day 2. Significant differences in the progressive motility of the semen stored at 5 °C and 16 °C were observed at each of the evaluation times. This reduced motility was consistent with decreased sperm mitochondrial transmembrane potential and oxidoreductive capability. We inferred that the cooling of boar semen to 5 °C increases the permeability of the sperm plasma membrane, which may not be revealed by SYBR-14/PI staining or HOS testing. The loss of boar sperm motility that occurs during hypothermic liquid preservation may be connected with alterations in the plasma membrane stability and disorders of mitochondrial transmembrane potential and oxidoreductive capability.

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“…However, inside the uterus, the anaerobic condition reduces the potential damage of reactive species (Foote et al 2002). ROS production is induced when semen is processed after ejaculation, especially due to contamination with leucocytes and the presence of spermatozoa with excess residual cytoplasm (Brouwers et al 2005). The spermatic cell also produces intracellular ROS due to flagellar activity (Gavella and Lipovac 1992).…”

Section: Discussionmentioning

confidence: 99%

Cooling of equine semen at 16ºC for 36h with the addition of cysteine in different concentrations

Oliveira¹,

Wolf²,

Viu³

et al. 2015

PHK

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Summary: Equine semen manipulation during the cooling process reduces sperm viability and fertility in consequence to, among others, membrane lipid peroxidation, because of the high content of polyunsaturated fatty acids, which makes cells highly susceptible to free radicals and reactive oxygen species. The objective ofthe present study was to evaluate the effect of in vitro addition of cysteine in four concentrations (0, 1, 1.5, and 2.5 mM) for cooling spermatozoa of 12 stallions at 16 ºC for 36 h. Evaluated variables were total motility, strength, viability and plasmatic and acrosomal membrane integrity at four different time points (0, 12, 24 and 36 h). With the exception of acrosomal integrity, it was verified a reduction in total motility, strength and plasmatic membrane integrity in all samples, during cooling. In the evaluations at 36 h of cold storage, total motility (mot) and viability (viab) were higher in groups treated with 1mM (mot:46,5 ± 6,1/viab:76,5 ± 6,9) and 1.5 mM (mot:46,0 ± 4,6/viab:76,9 ± 3,7) cysteine, respectively, compared to control (mot:35,5 ±18,4/viab:68,1±13,4) and 2.5mM (mot:39,7 ±12,4/viab:66 ±17,2) (P < 0.05). As for strength (str) and plasmatic membrane integrity (plasm), 1 mM cysteine (str:3.6 ± 0.5/plasm:57.2 ± 9.5) showed better results compared to control (str:3.2 ±1.1/plasm:54.1±11.8), 1.5 mM (str:3.5 ± 0.6/plasm:52.2 ±13.3) and 2.5 mM (str:3.2 ±1.1/plasm:55.8 ±12.5) (P < 0.05). Regarding acrosomal membrane integrity, in general, there was no loss of integrity (70.5 ±10.4; 69.4 ± 4.4; 68.0 ± 7.2 and 70.3 ± 0.5), control, 1 mM, 1.5 mM and 2.5 mM respectively. The concentration of 1mM cysteine was more efficient for the protection of sperm cells in the commercial system of passive cooling at 16 ºC for 36 h, with higher values for total motility, strength, viability and plasmatic membrane integrity.Keywords: antioxidant / cooled semen / genetic resources / spermatozoon / stallion Die Kühlung von Sperma über 36 Std. auf 16°C mit Zusatz von Cystein in verschiedenen KonzentrationenDie Manipulation von Pferdesamen während des Kühlprozesses reduziert deren Lebensfähigkeit und Fruchtbarkeit. Dies geschieht unter Anderem wegen der membranen Lipidperoxidation sowie des hohen Gehalts an mehrfach ungesättigten Fettsäuren, der die Zellen sehr anfällig für freie Radikale und reaktive Sauerstoff-Spezies macht. Das Ziel der vorliegenden Studie ist es, die Wirkung von künstlicher Zugabe von Cystein in vier Konzentrationen (0, 1, 1,5 und 2,5 mM) zu Kühlsperma von 12 Hengsten bei 16°C für 36 Stunden zu beurteilen. Ausgewertete Variablen waren Motilität, Lebensfähigkeit sowie plasmatische und akrosomale Membran-Integrität zu vier verschiedenen Zeitpunkten. Mit Ausnahme der akrosomalen Integrität wurde eine Reduktion der Motilität, der plasmatischen Membran-Integrität während der Abkühlung in allen Proben festgestellt. Die Bewertung nach 36 Stunden Kühlung ergab, dass die gesamte Motilität (Mot) und Lebensfähigkeit (Lbf) jeweils höher in der mit 1 mM Cystein behandelten Gruppe (Mot:46,5 ± 6,1/Lbf...

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New assays for detection and localization of endogenous lipid peroxidation products in living boar sperm after BTS dilution or after freeze–thawing

Cited by 106 publications

References 56 publications

Lipid Peroxidation in Human Spermatozoa from Men with Genitourinary Infections

Lipid Peroxidation in Human Spermatozoa from Men with Genitourinary Infections

Storage temperature of boar semen and its relationship to changes in sperm plasma membrane integrity, mitochondrial membrane potential, and oxidoreductive capability

Cooling of equine semen at 16ºC for 36h with the addition of cysteine in different concentrations

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