Assessing equine sperm-membrane integrity

Lagares, M. A.; Petzoldt, R; Sieme, Harald; Klug, E

doi:10.1046/j.1439-0272.2000.00351.x

Cited by 20 publications

(11 citation statements)

References 11 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Within these temperature ranges the spermatozoa continue to metabolise in aerobic phase, keeping their vital processes (Mann, 1975). However, damage caused by cooling at 5 °C was observed in a reduction of total motility and percentage of sperm cells with functional plasma membrane (Lagares et al. , 2000).…”

Section: Introductionmentioning

confidence: 99%

Metabolic evaluation of cooled equine spermatozoa

Vasconcelos¹,

Santana

Santos

et al. 2010

Andrologia

Self Cite

View full text Add to dashboard Cite

Microscopy has been used in the routine evaluation of sperm metabolism. Nevertheless, it has limited capacity to preview male fertility. As calorimetry may be used to evaluate directly the metabolic activity of a biological system, the aim of this study was to use microcalorimetry as an additive method for sperm metabolism evaluation of cooled equine semen. Two ejaculates of four stallions were collected and motility, viability (eosin 3%) and membrane functional integrity (hyposmotic swelling test) of spermatozoa were evaluated. Sperm samples were processed following different protocols and the metabolism of these samples was accessed by calorimetry. Centrifugation is part of some of these processing protocols and although this procedure has been deleterious for sperm viability and plasma membrane integrity, no decrease in sperm motility was observed. Microcalorimetry was capable of detecting the positive effect of re-suspending the sperm pellet with Kenney extender. Thus, the use of microcalorimetry offered additional information for equine sperm metabolism evaluation and was efficient in detecting important information from sperm cell metabolism.

show abstract

Section: Introductionmentioning

confidence: 99%

Metabolic evaluation of cooled equine spermatozoa

Vasconcelos¹,

Santana

Santos

et al. 2010

Andrologia

Self Cite

View full text Add to dashboard Cite

show abstract

“…The HOST was performed using a 1:2 dilution (semen:distilled water), and osmolarity of solution was 100 mOsm/kg H 2 O‐1. Samples were incubated at 37°C for 8 min according to protocol adjusted for horses (Lagares, Petzoldt, Sieme, & Klug, ). Subsequently, the sample was placed on a microscope slide and covered with a cover slip and assessed by phase contrast microscopy (400×) and counting 100 sperm cells.…”

Section: Methodsmentioning

confidence: 99%

Response to cooling of pony stallion semen selected by glass wool filtration

et al. 2017

View full text Add to dashboard Cite

The aim of this study was to compare the sperm separation technique using filtration through glass wool compared with just diluted cooled semen. Eighteen ejaculates were collected from 6 pony stallions of the Brazilian pony breed. Evaluations were done on pH, osmolarity, total motility, membrane functionality (HOST), membrane integrity (CFDA/PI), morphology and mitochondrial viability (MTT) in fresh, 24 and 48 h of cooled semen at 5°C. After dilution, the half of the extended semen was cooled (control group). The other half was cooled after filtration trough glass wool (filtered group). Retained semen was considered the portion of cells that did not transpose glass wool barrier. Total motility from the control, filtered and retained groups after 24 h of cooling was 35.5%, 43.3% and 10% (p < .0001) respectively. Sperm membrane integrity percentage at the CFDA/PI test was 37.9%, 44.8% and 14.8% (p < .0001), on the control, filtered and retained groups respectively. The results confirmed that the passage of spermatozoa through glass wool increased the selection of spermatozoa from pony stallions with higher motility, mitochondrial viability and membrane integrity for cooling in milk extender up to 24 h. Moreover, it was not obtained higher sperm parameters to control after cooling 48 h under the conditions that the study was conducted.

show abstract

“…Spermatocrit (haematocrit) tubes filled with the pellets in swelling medium was then measured, using a haematocrit centrifuge set (micro‐centrifuge K80h; Wagtech, Birmingham, UK) (15 000 × g , 3 min). The standardization of percent spermatocrit values for statistical analysis was also carried out (Lagares et al. 1999).…”

Section: Methodsmentioning

confidence: 99%

Bull Spermatozoa under Mercury Stress

Arabi

2005

Reprod Domestic Animals

View full text Add to dashboard Cite

Defective sperm function is the most prevalent cause of male infertility and is difficult to treat. This study was designed to evaluate the effect of mercuric chloride (HgCl2) at 50-300 micromol/l concentration range, in vitro, on the sperm membrane and DNA integrity, viability, reduced glutathione (GSH) content and acrosomal status of the bull spermatozoa. The samples were processed for sperm analyses using semen-diluting fluid [phosphate-buffered saline (PBS), pH 7.2]. I recorded a meaningful increase in the lipid peroxidation (LPO) rate and a drastic fall in the spermatocrit values under mercury additions, dominantly at 300 microM mercury concentration, indicating a deleterious effect of mercury on the sperm membrane intactness. There was also a strong negative correlation between LPO rate and percentage of viable spermatozoa (r = -0.9, p < 0.001). GSH content was significantly impaired. Data obtained from Comet assay [single-cell gel electrophoresis (SCGE)] technique revealed that mercury is capable of inducing DNA breaks in the sperm nuclei. Interestingly, 90% of DNA breaks were double-stranded. The correlation between LPO rate and percentage of DNA breaks was found to be 0.9 (p < 0.001). Results of the gelatin test indicate that mercury is capable of altering the integrity of acrosomal membranes, showing an abnormal acrosome reaction. In this regard, a strong correlation was found between LPO rate and percentage of halos (r = -0.9, p < 0.001). In conclusion, mercury proved to be a potential oxidant in the category of 'environmental factors' to bull spermatozoa. Hence, considering the widespread use of mercury and its compounds, these metals should be regarded with more concern.

show abstract

Assessing equine sperm-membrane integrity

Cited by 20 publications

References 11 publications

Metabolic evaluation of cooled equine spermatozoa

Metabolic evaluation of cooled equine spermatozoa

Response to cooling of pony stallion semen selected by glass wool filtration

Bull Spermatozoa under Mercury Stress

Contact Info

Product

Resources

About