A new method to analyze boar sperm DNA fragmentation under bright-field or fluorescence microscopy

Enciso, María; López‐Fernández, Carmen; Fernández, José Luis Fernández; Garcia, Pedro Castillo; Gosálbez, Altea; Gosálvez, Jaime

doi:10.1016/j.theriogenology.2005.05.044

Cited by 91 publications

(83 citation statements)

References 5 publications

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“…Similar results have been reported in previous research in boars (Boe-Hansen et al, 2005;De Ambrogi et al, 2006;Enciso et al, 2006) and in horses (López-Fernández et al, 2007).…”

Section: Discussionsupporting

confidence: 92%

“…Once in the laboratory, the following characteristics were evaluated: Sperm concentration (SC) (Neubauer chamber); Viability: sperm viability (eosinnigrosin); Sperm DNA fragmentation (DFI, %) (Halomax kit; Halotech, Madrid, Spain); and Sperm morphology, using primary abnormalities (PA, %; abnormalities of the head, intermediate pieces and insertion of the queue), secondary abnormalities (SA, %; other sperm abnormalities), and total abnormalities (TA, % = PA% + SA%). Sperm DNA fragmentation was evaluated using fluorescence microscopy (Olympus fluorescence microscope BX41TF, Olympus Corporation, Tokyo, Japan) according to the methodology, and Sperm Chromatin Dispersion test, described by Enciso et al (2006). The sperm DNA fragmentation was calculated as the percentage of damaged spermatozoa and the sperm abnormalities as the percentage of abnormal cells, both in the sample estimated by examining 200 spermatozoa.…”

Section: Methodsmentioning

confidence: 99%

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Seasonal variation in sperm characteristics of boars in southern Uruguay

2015

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-The objective of this study was to evaluate the effects of season, natural photoperiod, and room temperature at the housing facility on boar semen characteristics in Uruguay (34º66'S; 56º29'W). For this purpose, 117 ejaculates, obtained from eight adult males collected through 12 consecutive months, were assessed for sperm viability, DNA integrity, abnormalities (total, primary, and secondary), ejaculate volume, and sperm concentration. Viability, total and primary abnormalities, volume, and sperm concentration were affected by season. Sperm viability, volume, and sperm concentration were affected by natural photoperiod. In general, autumn and the decreasing photoperiod had a negative impact on most of the semen characteristics, except for volume. Housing temperature did not affect semen characteristics. In boars living in temperate climates, semen quality is negatively affected during autumn and is related to photoperiod changes; however, the effects of temperature changes in housingdo not affect these seminal characteristics. In this scenario, seasonal differences in semen quality may have a negative effect on sow fertilization. Consequently, semen quality control especially during autumn is imperative for the best boar selection to be used for insemination purposes. Seasonal differences in semen quality may have a negative effect on sow reproductive performance. This issue will be addressed in a future investigation.

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“…Similar results have been reported in previous research in boars (Boe-Hansen et al, 2005;De Ambrogi et al, 2006;Enciso et al, 2006) and in horses (López-Fernández et al, 2007).…”

Section: Discussionsupporting

confidence: 92%

Section: Methodsmentioning

confidence: 99%

Seasonal variation in sperm characteristics of boars in southern Uruguay

2015

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“…En nuestro estudio, esta quinta categoría no fue observada con ninguna de las concentraciones de ME ensayadas, posiblemente porque el protocolo utilizado es similar al método original de Fernández et al, (donde los autores también observaron pocos espermatozoides degradados) o porque se usó mercaptoetanol en lugar de ditiotreitol en la solución de lisis. Inicialmente, Enciso et al, (2006) ensayaron en espermatozoides de cerdo el kit de SCD desarrollado para humanos (Halosperm® kit), que incluye una incubación ácida previa a la lisis. Observaron imágenes similares a aquellas observadas en humanos (ausencia de halo o halos pequeños en los espermatozoides con fragmentación de su ADN y halos de dispersión alrededor de un "core" central en espermatozoides con su ADN intacto).…”

Section: Revisión Del Temaunclassified

“…Por consiguiente, propusieron una variante del kit de SCD para utilizar en espermatozoides de cerdo (Sperm-Sus-Halomax®), que incluye una breve incubación en una solución de lisis (sin una incubación ácida previa), obteniendo núcleos con extensos halos de dispersión de la cromatina en los espermatozoides con ADN fragmentado. Así, en la especie porcina, los núcleos espermáticos con ADN intacto (ausencia de fragmentación) no muestran o sólo presentan una mínima relajación de los "loops" de ADN cercanos al área de la cola espermática (Enciso et al, 2006). Patrones similares a los observados en cerdo fueron descritos en ratón (Rodríguez et al, 2005), toro (García-Macías et al, 2007;González-Marín et al, 2011) y carnero (López-Fernández et al, 2008a.…”

Section: Revisión Del Temaunclassified

Evaluación de la calidad del ADN espermático en camélidos sudamericanos y otras especies domésticas

Carretero¹,

Giuliano²,

Neild³

2017

SPERMOVA

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RESUMENLas anormalidades en el material genético pueden expresarse por defectos en la maduración o condensación nuclear, rupturas del ADN y anormalidades cromosómicas. Los parámetros de rutina utilizados para evaluar el semen (movilidad espermática, concentración, viabilidad, funcionalidad de membranas y morfología) no reflejan la calidad del ADN espermático. Esto cobra importancia para las técnicas de reproducción asistida, particularmente en la inyección intracitoplasmática de un espermatozoide en la que espermatozoides con genomas anormales pueden alcanzar el material genético del ovocito. Se postula que tres factores pueden estar involucrados en la etiología del daño del ADN: estrés oxidativo, aberraciones en el empaquetamiento de la cromatina y apoptosis. Varias técnicas han sido utilizadas para estudiar los defectos del ADN. A modo general, las técnicas de evaluación de ADN pueden ser divididas en 2 grupos: aquellas que evalúan el grado de condensación o compactación de la cromatina y aquellas que miden el grado de fragmentación del ADN. Muchas de estas técnicas son laboriosas, costosas y algunas dependen de enzimas cuya actividad y accesibilidad a las roturas de ADN pueden ser irregulares. En este artículo se comenta sobre el desarrollo y puesta a punto de varias de las técnicas de evaluación de la cromatina espermática en camélidos sudamericanos (CSA) y otras especies domésticas. Palabras clave: calidad de ADN, camélidos sudamericanos, espermatozoides ABSTRACTAbnormalities in sperm genetic material can be expressed as maturation or nuclear condensation defects, DNA damage or breakdown and chromosomal abnormalities. Routine semen evaluation (sperm motility, concentration, viability, membrane function and morphology) does not reflect sperm DNA quality. This becomes important when applying assisted reproductive techniques, especially intracytoplasmic sperm injection, in which sperm with abnormal genomes can reach the oocyte's genetic material. Three factors have been suggested to be involved in the etiology of DNA damage: oxidative stress, chromatin packaging anomalies and apoptosis. Various techniques have been used to study DNA defects. In general, DNA evaluation techniques can be divided into two groups: those that evaluate the degree of chromatin condensation or compactation and those that measure the degree of DNA fragmentation. Many of these techniques are laborious, costly and some depend on enzymes whose activity and accessibility to DNA fragments can be irregular. In this article, the development and setting up of various techniques to evaluate sperm chromatin in South American Camelids (SACs) and other domestic species is commented on.

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“…A simpler technique for monitoring DNA fragmentation in spermatozoa, the Sperm Chromatin Dispersion Assay, has been recently established (74). The major advantage of this method over others is that each test requires very small numbers of sperm; moreover, the test is very cost efficient.…”

Section: Chromatin Defectsmentioning

confidence: 99%

Cytometric solutions in veterinary andrology: Developments, advantages, and limitations

Petrunkina

Harrison²

2011

Cytometry Pt A

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Cytometric methodologies are becoming increasingly important in veterinary andrology as means of assessing sperm function. However, as yet, flow cytometric techniques in veterinary andrology have not kept up in sophistication with those in other areas of biology and medicine. In this brief review, we consider the present state of cytometry in andrological procedures for evaluating the fertility of domestic animal sires. We outline the aspects of sperm physiology, paying particular attention to the changes that take place during the process known as capacitation, which prepares the sperm for interaction with the egg. We then examine briefly but critically the cytometric techniques that are currently in commercial use or are being established in research laboratories for testing sperm characteristics. Current limitations and potential developments in semen assessment are discussed. Recent research knowledge offers possibilities for applying more subtle flow cytometric approaches to distinguish different levels of fertilizing potential in semen samples. For example, linking field fertility data to multiparametric kinetic studies of sperm capacitational changes rather than ''single parameter-single time point'' estimations may reveal that slower rather than rapid changes indicate high fertility. Moreover, the development of multicolor flow cytometric procedures as a means of evaluating multiple functional parameters in individual cells would reduce the uncertainties always inherent in predicting fertility from in vitro sperm evaluation tests. ' 2011 International Society for Advancement of Cytometry Key terms fertility; sperm evaluation; heterogeneity; subpopulations; multicolour cytometry THE main focus of an andrological assessment in veterinary medicine is to ascertain the fertility of a potential sire. Although the most obvious and indeed frequently used general veterinary test is to quantify the number of offspring born to the sire, this is a lengthy and very expensive process. For ''accuracy,'' many inseminations must be carried out, and results must await gestation and birth. Obviously, therefore, development of rapid tests that can be carried out in the laboratory has for long been important. However, as research into the reproductive process has advanced, it has become clear that the life of the fertilizing sperm between its release from the testis and its fusion with the egg is far from simple. Moreover, the population of sperm ejaculated into the female has been revealed as being heterogeneous in functional ability, even if morphologically normal [c.f. reviews (1,2)]. As a result of these findings, tests are being developed to examine functional ability within a heterogeneously responding population (3-5).Given that a normal male ejaculate contains many millions of individual sperm cells, it is natural that these tests are based on cytometry. In this short review, we will outline and comment upon a range of modern and still-developing cytometric methodologies in use or potentially useful for sperm...

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A new method to analyze boar sperm DNA fragmentation under bright-field or fluorescence microscopy

Cited by 91 publications

References 5 publications

Seasonal variation in sperm characteristics of boars in southern Uruguay

Seasonal variation in sperm characteristics of boars in southern Uruguay

Evaluación de la calidad del ADN espermático en camélidos sudamericanos y otras especies domésticas

Cytometric solutions in veterinary andrology: Developments, advantages, and limitations

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