Cluster analysis reveals a binary effect of storage on boar sperm motility function

2014 ◽  
Vol 26 (5) ◽  
pp. 623 ◽  
Author(s):  
Heiko Henning ◽  
Anna M. Petrunkina ◽  
Robin A. P. Harrison ◽  
Dagmar Waberski
Keyword(s):  
Cluster Analysis ◽  
Sperm Motility ◽  
Semen Parameters ◽  
Specific Response ◽  
Imaging Data ◽  
Sperm Analysis ◽  
Duration Of Storage ◽  
Boar Sperm ◽  
Storage Effects

Storage of liquid-preserved boar spermatozoa is associated with a loss of fertilising ability of the preserved spermatozoa, which standard semen parameters barely reflect. Monitoring responses to molecular effectors of sperm function (e.g. bicarbonate) has proven to be a more sensitive approach to investigating storage effects. Bicarbonate not only initiates capacitation in spermatozoa, but also induces motility activation. This occurs at ejaculation, but also happens throughout passage through the oviduct. In the present study we tested whether the specific response of boar sperm subpopulations to bicarbonate, as assessed by motility activation, is altered with the duration of storage in vitro. Three ejaculates from each of seven boars were diluted in Beltsville thawing solution and stored at 17°C. Only minor changes in the parameters of diluted semen were revealed over a period of 72 h storage. For assessment of bicarbonate responses, subsamples of diluted spermatozoa were centrifuged through a discontinuous Percoll gradient after 12, 24 and 72 h storage. Subsequently, spermatozoa were incubated in two Ca2+-free variants of Tyrode’s medium either without (TyrControl) or with (TyrBic) 15 mM bicarbonate, and computer-aided sperm analysis motility measurements were made. Cluster analysis of imaging data from motile spermatozoa revealed the presence of five major sperm subpopulations with distinct motility characteristics, differing between TyrBic and TyrControl at any given time (P < 0.001). Although there was an increasing loss of motility function in both media, bicarbonate induced an increase in a ‘fast linear’ cohort of spermatozoa in TyrBic regardless of storage (66.4% at 12 h and 63.9% at 72 h). These results imply a binary pattern in response of sperm motility function descriptors to storage: although the quantitative descriptor (percentage of motile spermatozoa) declines in washed semen samples, the qualitative descriptor (percentage of spermatozoa stimulated into fast linear motion by bicarbonate) is sustained independent of the duration of storage.

scholarly journals Effects of Astaxanthin on Miniature Pig Sperm Cryopreservation

2018 ◽  
Vol 2018 ◽  
pp. 1-9 ◽  
Author(s):  
Eunjoo Lee ◽  
Daeyoung Kim
Keyword(s):  
Sperm Motility ◽  
Semen Parameters ◽  
Control Group ◽  
Computer Assisted ◽  
Miniature Pig ◽  
Oxidation Stress ◽  
Sperm Analysis ◽  
Semen Parameter ◽  
Boar Sperm ◽  
Sperm Plasma Membrane

The purpose of this study is to evaluate the effects of astaxanthin added to freezing buffer on semen parameters, total sperm oxidation stress after postthawing of boar sperm, and lipid peroxidation (LPO) which is caused by reactive oxygen species (ROS) in sperm membrane. Varying concentrations of astaxanthin (0, 10, 50, 100, and 500 μM) were used in the freezing buffer during cryopreservation to protect the DNA of thawed miniature pig sperm. Semen parameter was measured using computer-assisted sperm analysis (CASA) for sperm motility, and then ROS rate and oxidative stress of boar sperm were determined using fluorescence-activated cell sorting (FACS). Sperm motility was higher (p<0.05) in the astaxanthin group than in the control group. Sperm motility and the number of progressive motile sperm were higher (p<0.05) in the astaxanthin 500 μM group than in the control group. In ROS evaluation, the astaxanthin group had lower intracellular O2 and H2O2 in viable sperm. Yo-Pro-I/HE and PI/H2DCFDA staining as revealed using flow cytometry was lower in astaxanthin groups than in the other groups. As a result, we found that astaxanthin could protect the sperm plasma membrane from free radicals and LPO during boar sperm postthawing.

scholarly journals Antioxidant Effect of a Polyphenol-Rich Murtilla (Ugni molinae Turcz.) Extract and Its Effect on the Regulation of Metabolism in Refrigerated Boar Sperm

2019 ◽  
Vol 2019 ◽  
pp. 1-15 ◽  
Author(s):  
Ignacio Jofré ◽  
Magdalena Cuevas ◽  
Leticia Signori de Castro ◽  
João Diego de Agostini Losano ◽  
Mariana Andrade Torres ◽  
...  
Keyword(s):  
Lipid Peroxidation ◽  
Sperm Motility ◽  
Sperm Quality ◽  
Membrane Damage ◽  
Antioxidant Effect ◽  
Boar Sperm ◽  
Fertilization Capacity ◽  
Ugni Molinae

The production of reactive oxygen species (ROS) in boar spermatozoa increases in refrigeration; this can have an impact on sperm quality and fertilization capacity. We evaluated the effect of polyphenol-rich aqueous extract of murtilla (Ugni molinae Turcz) on boar sperm stored at 17°C in order to reduce oxidative stress and improve sperm quality in the long term. Five experiments were performed: first, characterization of the polyphenol content from five genotypes of murtilla; second, determination of the genotype with the best antioxidant effect (MT-Ex); third, the antioxidant capacity on O2- and lipid peroxidation; fourth, the influence of MT-Ex on motility, calcium movement, cAMP, and metabolic parameters; and fifth, analysis of long-term refrigeration. The average phenolic content was 344 ppm; gallic acid, catechin, quercetin, myricetin, and kaempferol were detected. All extracts evaluated presented a concentration-dependent antioxidant effect. MT-Ex reduces intracellular O2-/peroxides but low lipid peroxidation. MT-Ex in nonstimulated ROS conditions reduces sperm motility, mitochondrial membrane potential, cAMP, and ATP, but the succinate dehydrogenase activity remained normal; also, we observed a reduction in calcium movement in in vitro sperm capacitation. The long-term analyses showed that MT-Ex improved sperm motility decay and reduced membrane damage and ROS at 168 h. Based on this study, we propose MT-Ex as a supplement in semen extenders.

scholarly journals Exosomes derived from HEK293T cells interact in an efficient and noninvasive manner with mammalian sperm in vitro

Nanomedicine ◽  
2020 ◽  
Vol 15 (20) ◽  
pp. 1965-1980
Author(s):  
Teresa Vilanova-Perez ◽  
Celine Jones ◽  
Stefan Balint ◽  
Rebecca Dragovic ◽  
Michael L Dustin ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Mitochondrial Membrane ◽  
Membrane Integrity ◽  
Computer Assisted ◽  
Sperm Function ◽  
Sperm Analysis ◽  
Mammalian Sperm ◽  
Boar Sperm ◽  
Computer Assisted Sperm Analysis

Aim: To investigate exosomes as a noninvasive delivery tool for mammalian sperm. Materials & Methods: Exosomes were isolated from HEK293T cells and co-incubated with boar sperm in vitro. Results: Internalized exosomes were detected within 10 min of co-incubation. Computer-assisted sperm analysis and flow cytometry demonstrated that even after 5-h of exposure to exosomes, there were no significant deleterious effects with regard to sperm motility, viability, membrane integrity and mitochondrial membrane potential (p > 0.05), thus indicating that exosomes did not interfere with basic sperm function. Conclusion: HEK293T-derived exosomes interacted with boar sperm without affecting sperm function. Exosomes represent a versatile and promising research tool for studying sperm biology and provide new options for the diagnosis and treatment of male infertility.

Epididymal constituents and related substances in the storage of spermatozoa: a review

1993 ◽  
Vol 5 (6) ◽  
pp. 601 ◽  
Author(s):  
BP Setchell ◽  
LG Sanchez-Partida ◽  
A Chairussyuhur
Keyword(s):  
Body Temperature ◽  
Sperm Motility ◽  
Related Substances ◽  
Duration Of Storage ◽  
Available Information ◽  
Mammalian Spermatozoa

In vertebrate animals, the duration of storage of viable spermatozoa in the epididymis varies from a few hours in birds to many months in reptiles and bats. The available information on the unusual composition of the epididymal luminal fluid is summarized, and the effect of the various constituents on sperm motility is described. Spermatozoa would probably be best stored in an immotile state and some constituents of epididymal luminal fluid may be able to inhibit the motility of mammalian spermatozoa during storage in vitro. If this effect can then be removed at the time of insemination, by changing the spermatozoa to a different medium, such a procedure may allow storage of spermatozoa at room or even body temperature for extended periods.

59 EFFECT OF STORAGE DURATION ON POST-THAW PARAMETERS OF BULL SEMEN

2013 ◽  
Vol 25 (1) ◽  
pp. 177
Author(s):  
D. B. Carwell ◽  
B. R. Scott ◽  
J. Len ◽  
H. Blackburn ◽  
K. R. Bondioli ◽  
...  
Keyword(s):  
Sperm Motility ◽  
Sperm Concentration ◽  
Time Frame ◽  
Water Bath ◽  
Semen Parameters ◽  
Storage Duration ◽  
Bovine Semen ◽  
Microcentrifuge Tube ◽  
Duration Of Storage ◽  
Time Frames

It has been proposed that once good-quality sperm are collected, extended, and cryopreserved that the post-thaw quality will remain high regardless of the duration of storage. A previous study has suggested that there is no effect on post-thaw sperm motility of bovine semen stored in liquid nitrogen up to 13 years. However, no studies have reported the effect of extended storage of bovine semen in LN2. In this study, cryopreserved semen from Angus bulls (n = 25) was utilized. Time frames were assigned to each bull based on the year of collection and cryopreservation: time frame 1 (1960–1975), time frame 2 (1976–1991), and time frame 3 (1992–2006). Due to differences in packaging methods across time periods (glass ampules and plastic straws), thawing methods utilized were different and based on the packaging type. Semen packaged in glass ampules (n = 7) were thawed in a 37°C water bath for 80 s, while those in plastic straws (n = 18) were thawed in a 37°C water bath for 30 s. Once thawed all packages were wiped dry and either scribed (glass ampule) or the end cut (plastic straw) and the semen expelled into a 1.5-mL microcentrifuge tube and maintained at 37°C for evaluation. For sperm motility, a 20-µL sample was placed onto a prewarmed microscope slide for analysis. Sperm motility parameters (total motility and progressive motility) were evaluated by 2 experienced technicians and averaged. A hemocytometer was used to determine sperm concentration in a final dilution of 1 : 100. An eosin/nigrosin stain was used to determine sperm morphology. Parameters were analyzed using one-way ANOVA. There was no difference in sperm concentration between time frames 1 and 2 (53 × 106 mL–1 and 59 × 106 mL–1, respectively) or time frames 1 and 3 (53 × 106 mL–1 and 37 × 106 mL–1, respectively). However, time frame 2 exhibited a higher (P < 0.05) sperm concentration compared with time frame 3 (59 × 106 mL–1 v. 37 × 106 mL–1, respectively). There were no differences among time frames 1, 2, or 3 for total (42, 51, and 55%, respectively) and progressive sperm motility (29, 38, and 41%, respectively). Similarly, there were no differences among time frames 1, 2, and 3 for percent normal sperm (80 ± 3.6, 76 ± 5.0, and 71 ± 4.0%, respectively) and abnormal sperm (19 ± 3.6, 23 ± 5.0, and 28 ± 4.0%, respectively). Furthermore, time frames 1, 2, and 3 did not exhibit a difference in primary (9 ± 2.5, 7 ± 2.6, and 6 ± 1.0%, respectively), secondary (2 ± 0.8, 5 ± 1.4, 6 ± 2.0%, respectively), or tertiary abnormalities (7 ± 2.4, 10 ± 2.7, and 16 ± 2.7%, respectively). These data suggest that once good-quality bovine sperm is cryopreserved, the duration of storage (up to 43 years) has no effect on Angus bull post-thaw semen parameters.

Effects of permethrin, cypermethrin and 3-phenoxybenzoic acid on rat sperm motility in vitro evaluated with computer-assisted sperm analysis

2010 ◽  
Vol 24 (2) ◽  
pp. 382-386 ◽  
Author(s):  
Chen Yuan ◽  
Cheng Wang ◽  
Shu-Qin Gao ◽  
Tian-Tian Kong ◽  
Lei Chen ◽  
...  
Keyword(s):  
Sperm Motility ◽  
Computer Assisted ◽  
Sperm Analysis ◽  
Computer Assisted Sperm Analysis

scholarly journals The effect of extender, method of thawing, and duration of storage on in vitro fertility measures of frozen–thawed boar sperm

2015 ◽  
Vol 84 (3) ◽  
pp. 407-412
Author(s):  
R.V. Knox ◽  
J.M. Ringwelski ◽  
K.A. McNamara ◽  
M. Aardsma ◽  
M. Bojko
Keyword(s):  
Duration Of Storage ◽  
Boar Sperm

scholarly journals In Vitro Effects of Enterococcus Faecalis and Selected Biomolecules on the Motility of Rabbit Spermatozoa

2017 ◽  
Vol 66 (3-4) ◽  
pp. 22-31
Author(s):  
Eva Tvrdá ◽  
Michal Ďuračka ◽  
Marek Halenár ◽  
Attila Kántor
Keyword(s):  
Sperm Motility ◽  
Negative Impact ◽  
Selective Advantage ◽  
Positive Control ◽  
Negative Control ◽  
Biologically Active ◽  
Rapid Decline ◽  
Sperm Analysis ◽  
Rabbit Spermatozoa

SummaryThis study assessed the potential efficiency of selected biologically active substances on the motility behavior of rabbit spermatozoa subjected to in vitro induced E. faecalis contamination. Semen samples were collected from 10 male rabbits and the presence of E. faecalis was confirmed using MALDI-TOF Mass Spectrometry. For the in vitro experiments rabbit spermatozoa were resuspended in the presence of 0,3 McF E. faecalis and different concentrations of selected biomolecules (resveratrol - RES, quercetin - QUE, curcumin - CUR, epicatechin - EPI, isoquercitrin - IZO). Sperm motility was assessed using the computer-aided sperm analysis at 0h, 2h, 4h, 6h and 8h. The presence of E. faecalis significantly decreased the motility (P<0.001) when compared to the untreated Control starting at 2h and maintaining this negative impact throughout the entire in vitro culture. Meanwhile, the motility was significantly higher in the experimental samples subjected to E. faecalis together 5 μmol/L RES (P<0.05), 10 μmol/L QUE (P<0.05) as well as 1 μmol/L (P<0.01) and 10 μmol/L CUR (P<0.05) when compared to the Positive Control (4h). No biomolecule was able to maintain the motion comparable to the Negative Control, and none was effective against the rapid decline of sperm motility caused by the presence of E. faecalis during later stages of the in vitro experiment (6h and 8h). We may conclude that RES, QUE and CUR may provide a selective advantage to spermatozoa in the presence of E. faecalis, particularly during short-term rabbit semen handling.

scholarly journals IN VITRO EFFECTS OF ACINETOBACTER BAUMANNII AND SELECTED NATURAL BIOMOLECULES ON RABBIT SPERMATOZOA MOTILITY

AGROFOR ◽  
2018 ◽  
Vol 3 (1) ◽  
Author(s):  
Eva TVRDÁ ◽  
Michal ĎURAČKA ◽  
Attila KÁNTOR ◽  
Marek HALENÁR ◽  
Lukáš HLEBA
Keyword(s):  
Sperm Motility ◽  
Negative Impact ◽  
White Male ◽  
Selective Advantage ◽  
Rapid Decline ◽  
Sperm Analysis ◽  
Spermatozoa Motility ◽  
In Vitro Effects ◽  
Rabbit Spermatozoa

The aim of this study was to assess the potential efficiency of selected biologicallyactive substances on the motility behaviour of rabbit spermatozoa subjected to invitro induced A. baumannii contamination. The semen samples used for A.baumannii detection were collected from 10 New Zealand white male rabbits andthe presence of the bacterium was confirmed using MALDI-TOF MassSpectrometry. For the in vitro experiments rabbit spermatozoa were re-suspendedin PBS, containing mineral supplements, BSA and glucose in the presence of 3x105CFU A. baumannii and diverse concentrations of selected biomolecules (resveratrol- RES, quercetin - QUE, curcumin - CUR, epicatechin - EPI, isoquercitrin - ISO).The sperm motility was assessed using the computer-aided sperm analysis at 0h,2h, 4h and 6h. A. baumannii significantly decreased the sperm motility (P<0.001)at Time 2h and maintained this negative impact throughout the in vitro culture.Meanwhile, the motility at Time 2h was significantly higher in the samples subjectedto A. baumannii together with 10 μmol/L RES (P<0.01); 5, 10 and 50 μmol/L QUE(P<0.001); 1 μmol/L CUR (P<0.05); 10, 50 and 100 μmol/L EPI (P<0.01) as well as 50μmol/L (P<0.05) and 100 μmol/L ISO (P<0.001) in comparison to the control exposedto the bacterium exclusively. After 4h, the motility remained significantly higher in thegroups co-treated with the inoculum and 10 μmol/L RES (P<0.05), 50 μmol/L QUE(P<0.05) as well as 50 μmol/L EPI (P<0.05) when compared to the positive control.Nevertheless, none of the biomolecules was effective against the rapid decline ofsperm motility caused by A. baumannii during later stages of the experiment (Time6h). Based on these results, one can conclude that RES, QUE and EPI exhibitantibacterial properties providing a selective advantage to spermatozoa in thepresence of A. baumannii, particularly during short-term rabbit semen handling.

scholarly journals Effects of zearalenone, α-zearalenol, and genistein on boar sperm motility in vitro

2017 ◽  
Vol 62 (No. 10) ◽  
pp. 435-445 ◽  
Author(s):  
A. Krejcárková ◽  
P. Folková ◽  
O. Šimoník ◽  
M. Šašková ◽  
R. Krejčířová ◽  
...  
Keyword(s):  
Sperm Motility ◽  
Computer Assisted ◽  
Sperm Analysis ◽  
Toxicological Research ◽  
Computer Assisted Sperm Analysis ◽  
In Vitro Effects ◽  
Dose Dependent Increase ◽  
Dose Dependent

Genistein (GEN) and zearalenone (ZEA), environmental oestrogens commonly present in feedstuff for pigs, are known for their effects on reproductive functions. The aim was to verify the in vitro effects of 0.5–20 µM concentrations of GEN, ZEA and its metabolite α-zearalenol (α-ZOL) on pig sperm motility. A dose-dependent increase of the immotile sperm amount against fast and medium-fast sperm clusters was observed with all three oestrogens from the lowest concentrations tested. Individual CASA (computer-assisted sperm analysis) parameters of motile sperms seemed to be less sensitive indicators. This should be considered especially in toxicological research on a sperm model. Background of inconsistencies in to date-published papers is discussed. The results shift the effective concentrations of ZEA, α-ZOL, and GEN to values achievable in vivo and raises the questions of risk assessment of these compounds in pig reproduction.

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