Metabolomic Analysis and Identification of Sperm Freezability Biomarkers in Boar Seminal Plasma

2021 ◽  
Author(s):  
Yu-ting Zhang ◽  
Han-lin Liang ◽  
Yan Liu ◽  
Meng Zhao ◽  
Qian-qian Xu ◽  
...  
Keyword(s):  
Small Molecules ◽  
Aspartic Acid ◽  
Seminal Plasma ◽  
Freezing Process ◽  
Accessory Sex Glands ◽  
Plasma Metabolome ◽  
Boar Sperm ◽  
Boar Semen ◽  
The Difference ◽  
Sperm Functions

Abstract Background: In the freezing process of boar sperm, there are obvious differences of freezability between individuals. Studies suggest that specific freezability markers might be useful in good (GFE) and poor freezabitity ejaculates (PFE) selection prior to cryopreservation. Some potential markers of boar sperm freezability have been found from spermatozoa, little attention has been paid to seminal plasma. The seminal plasma is composed of secretions from testis, epididymis, and accessory sex glands, and the exposure of spermatozoa to small molecules such as metabolites can affect the sperm functions. However, details and significance of the seminal plasma metabolome related to boar sperm freezability are unknown. Therefore, the main aim of this study was to explore the difference in the metabolic level of seminal plasma between boars with differential freezability, and to explore the biomarkers of semen freezing tolerance. Results: A total of 953 metabolites were identified in boar semen plasma by UHPLC-qTOF-MS analysis, and 50 metabolites show significant change between GFE group and PFE group. Further, twelve metabolites were subjected to metabolic target analysis and three metabolites (D-Aspartic acid, N-Acetyl-L-glutamate (NAG), and Inosine) show differences.Conclusions: There is significant differece on metabolome of seminal plasma between GFE and PFE individuals. The D-Aspartic acid, NAG, and Inosine in seminal plasma may be potential markers for assessing sperm cryopreservation resistance in boars.

scholarly journals Metabolomic Analysis and Identification of Sperm Freezability-Related Metabolites in Boar Seminal Plasma

Animals ◽  
2021 ◽  
Vol 11 (7) ◽  
pp. 1939
Author(s):  
Yuting Zhang ◽  
Hanlin Liang ◽  
Yan Liu ◽  
Meng Zhao ◽  
Qianqian Xu ◽  
...  
Keyword(s):  
Small Molecules ◽  
Aspartic Acid ◽  
Seminal Plasma ◽  
Metabolomic Analysis ◽  
Sperm Cryopreservation ◽  
Accessory Sex Glands ◽  
Plasma Metabolome ◽  
Significant Difference ◽  
Boar Sperm ◽  
Boar Semen

Some potential markers of boar sperm freezability have been found in spermatozoa, but little attention has been paid to seminal plasma. The seminal plasma is composed of secretions from the testis, epididymis, and accessory sex glands. The exposure of spermatozoa to small molecules such as metabolites can affect sperm function. However, details and significance of the seminal plasma metabolome related to boar sperm freezability are unknown. Therefore, the main aim of this study was to explore the differences in the metabolic level of seminal plasma between boars with differential freezability and to explore the candidate biomarkers of semen freezability. A total of 953 metabolites were identified in boar semen plasma by UHPLC-qTOF-MS analysis, and 50 metabolites showed significant change between the GFE group and PFE group. Further, twelve metabolites were subjected to metabolic target analysis, and three metabolites (D-aspartic acid, N-acetyl-L-glutamate (NAG), and inosine) showed differences. In conclusion, there is significant difference in the metabolome of seminal plasma between GFE and PFE individuals. D-aspartic acid, NAG, and inosine in seminal plasma may be potential markers for assessing sperm cryopreservation resistance in boars.

52 Difference of Seminal Plasma Proteins in Good- and Poor-Freezability Boar Ejaculates

2018 ◽  
Vol 30 (1) ◽  
pp. 165 ◽  
Author(s):  
J. Rungruangsak ◽  
J. Suwimonteerabutr ◽  
S. Asawakarn ◽  
K. Buranaamnuay ◽  
N. Chantaravisoot ◽  
...  
Keyword(s):  
Seminal Plasma ◽  
Plasma Proteins ◽  
Protein Extraction ◽  
Normal Morphology ◽  
Progressive Motility ◽  
Semen Collection ◽  
Seminal Plasma Proteins ◽  
Boar Semen ◽  
One Way Anova ◽  
Sperm Functions

Seminal plasma is the semen components that maintain sperm metabolism, pH and osmolality. Fibronectin (FN1) and glutathione peroxidase (GPX5) are the seminal plasma proteins that play an important role on the boar sperm functions. The purpose of the present study was to determine the differences in GPX5 and FN1 contents in the boar semen having good, moderate, and poor freezability. A total of 38 ejaculates from 25 boars with proven fertility were included in the experiment. All the ejaculates included in the study had >70% subjective motility, >75% normal morphology, >75% sperm viability, and volume >100 mL per ejaculate. The semen was collected through semen collection bag with filter and split into 2 portions. The first portion was prepared for the evaluation of seminal plasma proteins (i.e. GPX5 and FN1) and the second portion was cryopreserved and evaluated for post-thaw semen qualities. The seminal plasma samples were collected in cryotube and plug into liquid nitrogen. The samples was stored at –80°C before protein extraction. After thawing, the ejaculates were classified into 3 groups according to their post-thawed sperm motility: poor (14.6 ± 3.9%), modersate (28.5 ± 4.1%), and good (64.0 ± 8.7%) freezability. The amounts of GPX5 and FN1 proteins were evaluated through Western blot analysis. The normalized quantity of proteins was compared among groups by one-way ANOVA. The normalized level of FN1 in seminal plasma was higher in good- than in poor-freezability groups (8.0 ± 0.8% v. 5.7 ± 0.7%, respectively; P < 0.05), but did not differ significantly compared with that of the moderate-freezability group (7.5 ± 0.8%; P > 0.05). The levels of GPX5 in good-, moderate-, and poor-freezability groups were 14.7 ± 3.0%, 16.8 ± 3.2%, and 10.9 ± 3.0%, respectively (P > 0.05). The level of FN1 in seminal plasma was significantly correlated with the post-thaw sperm progressive motility (r = 0.38, P = 0.01), total motility (r = 0.37, P = 0.02), and the proportion of bent tail sperm (r = –0.33, P = 0.04). The level of GPX5 was not correlated with any of the post-thaw sperm qualities (P > 0.05). However, the levels of GPX5 was positively correlated with FN1 (r = 0.40, P = 0.01, n = 38). It can be concluded that FN1 in seminal plasma can be used as a marker of sperm freezability in boar.

Metabolomic signature of spermatozoa established during holding time is responsible for differences in boar sperm freezability

2021 ◽  
Author(s):  
Mariana A Torres ◽  
Ana Carolina Pedrosa ◽  
Francisco José Novais ◽  
Diego V Alkmin ◽  
Bruce R Cooper ◽  
...  
Keyword(s):  
Seminal Plasma ◽  
Sperm Quality ◽  
Membrane Integrity ◽  
Membrane Lipid ◽  
Holding Time ◽  
Glutathione Metabolism ◽  
Mitochondrial Potential ◽  
Metabolic Markers ◽  
Boar Sperm ◽  
The Difference

Abstract Holding at room temperature is the first step in most boar semen cryopreservation protocols. It is well accepted that a holding time (HT) of 24 h increases sperm cryotolerance. However, the effect of HT on ejaculates with different freezability is not entirely clear. The aim of this study was to understand how HT influences spermatic and seminal plasma metabolite profiles of boar ejaculates and how these possible changes affect freezability. Twenty-seven ejaculates were collected and extended to 1:1 (v: v) with BTS and split into two aliquots. The first aliquot was cryopreserved without holding time (0 h), and the second was held at 17°C for 24 h before cryopreservation. Spermatozoa and seminal plasma were collected by centrifugation at two times, before HT (0 h) and after HT (24 h), and subsequently frozen until metabolite extraction and UPLC–MS analysis. After thawing, the semen samples were evaluated for kinetics, membrane integrity, mitochondrial potential, membrane lipid peroxidation, and fluidity. The ejaculates were then allocated into two phenotypes (good ejaculate freezers [GEF] and poor ejaculate freezers [PEF]) based on the percent reduction in sperm quality (%RSQ) as determined by the difference in total motility and membrane integrity between raw and post-thaw samples cryopreserved after 24 h of HT. The metabolic profile of the seminal plasma did not seem to influence ejaculate freezability, but that of the spermatozoa were markedly different between GEF and PEF. We identified a number of metabolic markers in the sperm cells (including inosine, hypoxanthine, creatine, ADP, niacinamide, spermine, and 2-methylbutyrylcarnitine) that were directly related to the improvement of ejaculate freezability during HT; these were components of metabolic pathways associated with energy production. Furthermore, PEF showed an up-regulation in the arginine and proline as well as the glutathione metabolism pathways. These findings help to better understand the effect of holding time on boar sperm freezability and propose prospective metabolic markers that may predict freezability; this has implications in both basic and applied sciences.

scholarly journals Thawing boar semen in the presence of seminal plasma improves motility, modifies subpopulation patterns and reduces chromatin alterations

2017 ◽  
Vol 29 (8) ◽  
pp. 1576 ◽  
Author(s):  
Rocío Fernández-Gago ◽  
Manuel Álvarez-Rodríguez ◽  
Marta E. Alonso ◽  
J. Ramiro González ◽  
Beatriz Alegre ◽  
...  
Keyword(s):  
Seminal Plasma ◽  
Chromatin Structure ◽  
Sperm Chromatin ◽  
Chromatin Compaction ◽  
Sperm Analysis ◽  
Positive Effects ◽  
Fragmentation Index ◽  
Non Linear ◽  
Boar Semen ◽  
Dna Fragmentation Index

Seminal plasma could have positive effects on boar semen after thawing. In the present study we investigated changes in the motility and chromatin structure in spermatozoa over 4 h incubation (37°C) of boar semen thawed in the presence of 0%, 10% or 50% seminal plasma from good-fertility boars. Cryopreserved doses were used from seven males, three of which were identified as susceptible to post-thawing chromatin alterations. Motility was analysed by computer-aided sperm analysis every hour, and data were used in a two-step clustering, yielding three subpopulations of spermatozoa (slow non-linear, fast non-linear, fast linear). Chromatin structure was analysed using a sperm chromatin structure assay and flow cytometry to determine the DNA fragmentation index (%DFI) as a percentage, the standard deviation of the DFI (SD-DFI) and the percentage of high DNA stainability (%HDS), indicating chromatin compaction. Thawing without seminal plasma resulted in a rapid loss of motility, whereas seminal plasma helped maintain motility throughout the incubation period and preserved the subpopulation comprising fast and linear spermatozoa. The incidence of chromatin alterations was very low in samples from non-susceptible males, whereas samples from males susceptible to post-thawing chromatin alterations exhibited marked alterations in %DFI and %HDS. Seminal plasma partly prevented these alterations in samples from susceptible males. Overall, 50% seminal plasma was the most efficient concentration to protect motility and chromatin. Some changes were concomitant with physiological events reported previously (e.g., semen thawed with 50% seminal plasma increased the production of reactive oxygen species and yielded higher fertility after AI). Thawing in the presence of seminal plasma could be particularly useful in the case of samples susceptible to post-thawing chromatin damage.

Spermatozoa and seminal plasma small extracellular vesicles miRNAs as biomarkers of boar semen cryotolerance

2021 ◽  
Author(s):  
Ana Carolina Pedrosa ◽  
Mariana A. Torres ◽  
Diego V. Alkmin ◽  
Jorge E.P. Pinzon ◽  
Simone Maria M.K. Martins ◽  
...  
Keyword(s):  
Extracellular Vesicles ◽  
Seminal Plasma ◽  
Boar Semen

Seminal plasma proteins protect flow-sorted ram spermatozoa from freeze - thaw damage

2009 ◽  
Vol 21 (4) ◽  
pp. 571 ◽  
Author(s):  
T. Leahy ◽  
J. I. Marti ◽  
G. Evans ◽  
W. M. C. Maxwell
Keyword(s):  
Seminal Plasma ◽  
High Speed ◽  
Sperm Quality ◽  
Membrane Integrity ◽  
Weight Fraction ◽  
Protein Supplementation ◽  
Freezing Process ◽  
Spermatozoa Motility ◽  
Functional Integrity

Seminal plasma improves the functional integrity of compromised ram spermatozoa but has been reported to be toxic to sorted spermatozoa. The present study attempted to clarify this paradoxical effect and improve the functional integrity of spermatozoa following sorting and cryopreservation. The in vitro function of sorted spermatozoa (motility characteristics and membrane integrity) was examined after supplementation with differing concentrations and protein fractions of seminal plasma at various stages of the sorting and freezing process. For all experiments, spermatozoa (two males, n = four ejaculates per male) were processed through a high-speed flow cytometer before cryopreservation, thawing and incubation for 6 h (37°C). Supplementation of crude seminal plasma (CP), its low molecular weight fraction (LP; <10 kDa) or protein-rich fraction (SPP; >10 kDa), immediately before freezing improved the functional integrity of sorted spermatozoa compared with no supplementation (control), whereas supplementation after thawing had no effect for CP and LP. The protective effect of seminal plasma was not altered by increasing the amount of protein supplementation. No toxic effect of CP, SPP or LP was evident even when supplemented at high protein concentrations. It is concluded that seminal plasma protein, if added to ram spermatozoa after sorting and before freezing, can improve post-thaw sperm quality and consequently the efficiency of sorting. This effect is most likely related to protection of the spermatozoa during freeze–thawing.

scholarly journals Seminal plasma arising from the whole boar sperm-rich fraction increases the stability of sperm membrane after thawing1,2

2016 ◽  
Vol 94 (5) ◽  
pp. 1906-1912 ◽  
Author(s):  
M. A. Torres ◽  
G. M. Ravagnani ◽  
D. F. Leal ◽  
S. M. M. K. Martins ◽  
B. B. D. Muro ◽  
...  
Keyword(s):  
Seminal Plasma ◽  
Boar Sperm ◽  
Sperm Membrane ◽  
The Stability

Seasonal changes of steroid concentrations in seminal plasma of a European wild boar

1984 ◽  
Vol 107 (3) ◽  
pp. 425-427 ◽  
Author(s):  
D. Schopper ◽  
J. Gaus ◽  
R. Claus ◽  
H. Bader
Keyword(s):  
Wild Boar ◽  
Seminal Plasma ◽  
Seasonal Pattern ◽  
Plasma Concentrations ◽  
Boar Taint ◽  
Steroid Production ◽  
European Wild Boar ◽  
Clear Seasonal Pattern ◽  
Boar Semen ◽  
Low Levels

Abstract. The influence of season on testicular steroid production as a parameter of testicular function has been studied in a wild boar. Semen was collected once weekly while it served the dummy. In seminal plasma concentrations of the following steroids were determined by radioimmunoassay: unconjugated testosterone, conjugated testosterone, unconjugated total oestrogens, conjugated total oestrogens and 5α-androst-16-en-3-one ('boar-taint steroid'). All steroids showed a clear seasonal pattern with highest concentrations in autumn and early winter and low levels from January to July. Maxima during the rutting season were 10–25 times greater than average values out of season. During a 2-month-period (mid-July until mid-September) libido was abolished and the wild boar refused to mount the dummy. These results indicate that the seasonal variation in testicular steroid production by the wild boar, regulated by photoperiod, are similar to those of the domestic boar.

Absence of seminal plasma from sperm-rich fraction decreases boar sperm quality characteristics during the course of liquid storage

2018 ◽  
Vol 198 ◽  
pp. 20-26 ◽  
Author(s):  
D.F. Leal ◽  
M.A. Torres ◽  
G.M. Ravagnani ◽  
S.M.M.K. Martins ◽  
F.V. Meirelles ◽  
...  
Keyword(s):  
Seminal Plasma ◽  
Sperm Quality ◽  
Quality Characteristics ◽  
Liquid Storage ◽  
Boar Sperm
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