Subtle membrane changes in cryopreserved bull semen in relation with sperm viability, chromatin structure, and field fertility

Januškauskas, Aloyzas; Johannisson, Anders; Rodriguez‐Martinez, Heriberto

doi:10.1016/s0093-691x(03)00050-5

Cited by 228 publications

(173 citation statements)

References 46 publications

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“…There are numerous staining techniques for determination of membrane status (Gillian et al 2005), in particular, various fluorescently labeled lectins (Flesch et al 1998;Jankovičová et al 2008), SYBR-14 (Garner and Johnson 1995) or membrane destabilization marker -annexin V (Peňa et al 2003). It is hypothesized that the incidence of cells with PS translocation may indicate the cells with altered membrane function that will eventually undergo necrosis (Januskauskas et al 2003). Such PS translocation may be occurred in mature spermatozoa (Peňa et al 2005) and mostly is considered as an apoptosis harbinger.…”

Section: Discussionmentioning

confidence: 99%

Functional characteristics of ram cooling-stored spermatozoa under the influence of epidermal growth factor

Makarevich

Špaleková²,

Olexíková³

et al. 2011

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Abstract. The aim of the study was to examine the effects of epidermal growth factor (EGF) on ram sperm traits following hypothermic storage. Fresh ram ejaculates were diluted in Triladyl extender, pooled and divided into groups according to EGF doses added (0, 100, 200 or 400 ng/ml). Following 72-96 h storage at 4°C, the spermatozoa were stained for a plasma membrane integrity (PNA-FITC), membrane phosphatidylserine (PS) translocation (annexin V-Fluos) and apoptosis (Yo-Pro-1), and analyzed by fluorescent microscopy. Sperm motility was measured using computer-assisted sperm analysis (CASA) and sperm fertilizing ability was tested using in vitro penetration/fertilization test on bovine prematured oocytes. EGF increased sperm motility at all doses tested, decreased the proportion of spermatozoa with damaged plasma membrane (at 200 or 400 ng/ml), and decreased the proportion of apoptotic (Yo-Pro-1) spermatozoa when given at 200 or 400 (but not 100) ng/ml. The proportion of spermatozoa with PS translocations (8.5%) was affected by neither of the EGF concentrations tested. However, fertilizing ability of ram sperm in the in vitro test was not improved by EGF (200 ng/ml). In summary, EGF when given at higher concentrations improved sperm viability and motility after cooling storage, but these effects were not reflected in sperm fertilizing ability in vitro.

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Section: Discussionmentioning

confidence: 99%

Functional characteristics of ram cooling-stored spermatozoa under the influence of epidermal growth factor

Makarevich

Špaleková²,

Olexíková³

et al. 2011

gpb

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“…This combination can detect four categories of sperm populations: live, live early 'apoptotic', dead and late 'apoptotic', and late 'necrotic' cells. 125 The APO-BRDU kit to conduct TUNEL assay for determination of DNA fragmentation was found more effective than the Annexin/PI in bull spermatozoa, although detection of necrotic spermatozoa cannot be performed by the APO-BRDU kit. The TUNEL assay measures changes at the later stage, while Annexin V/PI measure early sperm apoptosis-like changes.…”

Section: Changes Induced During Capacitationmentioning

confidence: 91%

Flow cytometry for the assessment of animal sperm integrity and functionality: state of the art

Hossain¹,

Johannisson²,

Wallgren³

et al. 2011

Asian J Androl

141

120

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Flow cytometry is now a recognized methodology within animal spermatology, and has moved from being a research tool to become routine in the assessment of animal semen destined to breeding. The availability of 'bench-top' flow cytometers and of newer and versatile markers for cell structure and function had allowed the instrumentation to measure more sperm parameters, from viability to reactiveness when exposed to exogenous stimuli, and to increase our capabilities to sort spermatozoa for potential fertilizing capacity, or chromosomal sex. The present review summarizes the state of the art regarding flow cytometry applied to animal andrology, albeit keeping an open comparative intent. It critically evaluates the present and future capabilities of flow cytometry for the diagnostics of potential fertility and for the development of current reproductive technologies such as sperm freezing, sperm selection and sperm sorting. The flow cytometry methods will probably further revolutionize our understanding of the sperm physiology and their functionality, and will undoubtedly extend its application in isolating many uncharacterized features of spermatozoa. However, continuous follow-up of the methods is a necessity owing to technical developments and the complexity of mapping spermatozoa.

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“…Sperm cells with damaged plasma membrane become unable to accomplish oocyte fertilization Gadella, 2006). Januskauskas et al (2003) detected significant correlations between field fertility and PMI assessed by PI. Correa et al (1997) demonstrated a positive correlation with PMI and high fertility bulls.…”

Section: Resultsmentioning

confidence: 92%

“…Hence, multiple independent variables are often necessary in order to make a reasonably accurate prediction (UTT, 2016). For this reason, it is necessary to associate different semen quality tests so to improve the accuracy of a bull's reproductive potential (JANUSKAUSKAS et al, 2003;TARTAGLIONE;RITTA, 2004;THUNDATHIL, 2008;ARRUDA et al, 2011;OLIVEIRA et al, 2013;SELLEM et al, 2015;CELEGHINI et al, 2017).…”

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confidence: 99%

Assessment of different in vitro sperm challenges and in vivo fertility of bovine semen batches

Oliveira

Ribeiro

Silva

et al. 2017

Braz. J. Vet. Res. Anim. Sci.

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The aim of this work was to submit sperm cells to different laboratory challenges and to compare in vitro results with in vivo semen fertility. Four different batches from the same Brangus bull were used in a timed-AI program of 332 Brangus cows. Each batch (B) was submitted to the following procedure: semen sample was thawed at 36°C for 30 seconds (control). Sperm motility parameters, plasma membrane integrity, sperm morphology, and concentration were assessed. Then, an aliquot of thawed sample was incubated in a water bath at 45°C for 40 min (thermal challenge group; TCG) and another aliquot was centrifuged at 500 xg (Percoll gradient 45%/90%) for 15 min (centrifugation challenge group; CCG). Centrifuged semen was also submitted to another thermal challenge, being incubated (water bath) at 45°C for 40 min (centrifugation + thermal challenge group; CTCG). At the end of each challenge (CCG, TCG, and CTCG), the same laboratory tests used for control group were repeated. The following conception rates (CR) were observed for each batch: B1 = 48.9% (44/90); B2 = 44.2% (23/52); B3 = 55.5% (40/72); B4 = 43.2% (51/118); (p < 0.10). In the lab, B3 presented higher (p ≤ 0.05) progressive motility (PM) than B4 after thawing (control group) and after all sperm challenges (TCG, CCG, and CTCG). However, despite B3 and B4 having demonstrated a similar percentage of plasma membrane integrity (PMI) to the control group (B3 = 66.7 ± 1.3 and B4 = 65.2 ± 3.3), B3 demonstrated higher (P ≤ 0.05) percentage of PMI (37.2 ± 2.5) than B4 (26.7 ± 3.3) after passing through the most stressing in vitro challenge (CTCG). The semen batch presenting the highest resistance to in vitro challenges was the one that presented a trend for higher in vivo fertility, suggesting that submitting semen samples to laboratory challenges may be an interesting alternative for selecting batches with greater field fertility. Keywords: Conception rate. In vitro sperm resistance. Timed-AI. ResumoO objetivo deste estudo foi estressar células espermáticas em diferentes desafios laboratoriais e comparar os resultados in vitro com a fertilidade in vivo do sêmen. Quatro partidas de um mesmo touro Brangus foram utilizadas em um programa de IATF de 332 vacas Brangus. Cada partida foi submetida ao seguinte procedimento: a amostra de sêmen foi descongelada a 36°C por 30 segundos (grupo controle). Foram avaliados parâmetros de motilidade espermática (CASA), integridade da membrana plasmática (PMI), morfologia e concentração espermática. Em seguida, uma alíquota da amostra descongelada foi incubada em banho-maria a 45°C durante 40 minutos (grupo de desafio térmico, TCG) e outra alíquota foi centrifugada a 500 xg (gradiente de Percoll 45%/90%) durante 15 min (grupo desafio de centrifugação, CCG). Uma aliquota do sêmen centrifugado foi ainda submetida ao desafio térmico, sendo incubado a 45°C durante 40 min (grupo de desafio térmico + centrifugação, CTCG). No final de cada desafio (CCG, TCG e CTCG), os mesmos testes laboratoriais utilizados para o grupo de controle f...

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Subtle membrane changes in cryopreserved bull semen in relation with sperm viability, chromatin structure, and field fertility

Cited by 228 publications

References 46 publications

Functional characteristics of ram cooling-stored spermatozoa under the influence of epidermal growth factor

Functional characteristics of ram cooling-stored spermatozoa under the influence of epidermal growth factor

Flow cytometry for the assessment of animal sperm integrity and functionality: state of the art

Assessment of different in vitro sperm challenges and in vivo fertility of bovine semen batches

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Subtle membrane changes in cryopreserved bull semen in relation with sperm viability, chromatin structure, and field fertility

Cited by 228 publications

References 46 publications

Functional characteristics of ram cooling-stored spermatozoa under the influence of epidermal growth factor

Functional characteristics of ram cooling-stored spermatozoa under the influence of epidermal growth factor

Flow cytometry for the assessment of animal sperm integrity and functionality: state of the art

Assessment of different in vitro sperm challenges and in vivo fertility of bovine semen batches

Contact Info

Product

Resources

About

Functional characteristics of ram cooling-stored spermatozoa under the influence of epidermal growth factor

Functional characteristics of ram cooling-stored spermatozoa under the influence of epidermal growth factor