Transmembrane protein 95 as a promising molecular marker of ram sperm functionality

Theriogenology, Journal Year: 2025, Volume and Issue: unknown, P. 117440 - 117440

Published: April 1, 2025

The optimization of preservation protocols (refrigeration and freezing) in ovine species is necessary for a wider diffusion artificial insemination this species. Besides the ram sperm quality assays, characterization novel proteins could be crucial improving these employing biomarkers. protein transmembrane 95 (TMEM95) membrane associated with oocyte-sperm fusion previously described bull or mouse. However, has not yet been characterized until now. In work, different experimental groups based on functionality: capacitated, refrigerated at times (5 °C 24 h, 5 48 72 h), frozen-thawed samples were analyzed compared to initial (15 3 h) characterize expression its relationship other markers (motility, kinetic parameters, viability, apoptosis-like events, mitochondrial function, acrosome-reacted, zinc content as marker capacitation). addition, capacitation status was tested by Fluozin-3, fluorescent probe measuring used first time sperm. After induction, expected, acrosome reactive spermatozoa signature 2 significantly increased, while linearity (P < 0.05) decreased non-capacitated samples. Concerning TMEM95, profile increased after process, confirming status. Attending processes, semen progressively during liquid storage, significant decrease observed h according fast progressive motility linearity. TMEM95 showed same tendency, showing reduction respect control Finally, cryopreservation thawed suffered detrimental effect sample, concerting all studied parameters accomplished When we correlation markers, highest positive correlations low capacitated samples, such changes acrosome-reaction. On hand, between process suitable functionality). According our results, considered predictor early damage (cooling freezing), considering integrity

Language: Английский

---

Comparison of Post-Thaw Motility and In Vitro Fertility Between Ejaculated and Epididymal Semen, and Seminal cfDNA Characterization in Pantaneiro Bulls

Biology, Journal Year: 2025, Volume and Issue: 14(5), P. 465 - 465

Published: April 25, 2025

This study evaluated the post-thaw motility and in vitro fertility of ejaculated epididymal semen from Pantaneiro bulls characterized cell-free DNA (cfDNA) fresh seminal plasma. Semen five was collected via electroejaculation or post-mortem extraction. Fresh parameters cfDNA concentrations were assessed before cryopreservation. Post-thaw sperm kinetics using CASA at 0 6 h incubation, embryo development analyzed following IVF. Data ANOVA logistic regression. Ejaculate samples exhibited more morphological defects than (15.8% vs. 1.8%, p ≤ 0.05). Post-thaw, showed higher total (87.2% 32.4%) progressive (67.1% 14.4%) (p 0.05), (38.9% 11.0%, In did not differ significantly between (n = 525 oocytes) 500 groups terms cleavage (49.6% 44.2%) blastocyst formation on D7 (26.1% 22.2%, > concentration ranged 11.4 to 50.9 ng/µL. These findings indicate that retain high fertility. Additionally, characterization plasma contributes indigenous cattle preservation advances male research.

Language: Английский

---

Freezing–Thawing Procedures Remodel the Proteome of Ram Sperm before and after In Vitro Capacitation

International Journal of Molecular Sciences, Journal Year: 2019, Volume and Issue: 20(18), P. 4596 - 4596

Published: Sept. 17, 2019

Mammalian sperm must undergo a set of structural and functional changes collectively termed as capacitation to ensure successful oocyte fertilization. However, can be compromised by cryopreservation procedures, which alter the proteome longevity sperm. To date, how protein induced could affect acquisition fertilizing potential remains unexplored. The present study investigated profile ram during in vitro before after elucidate impact on at molecular level. Fresh cryopreserved were incubated under capacitating (CAP) non-capacitating (NC) conditions for 240 min. these four treatments was analyzed compared different incubation times using reverse phase liquid chromatography coupled mass spectrometry (RP-LC-MS/MS). comparison between fresh suggested that facilitated an apoptosis-stress response redox process, while CAP NC showed increased those biological processes associated with signaling, metabolism, motility, reproductive processes. In addition, 14 proteins related mitochondrial activity, recognition, spermatogenesis, underwent significant abundance over time when compared. Our results indicate disturbances may quality its specific machinery sustain conditions.

Language: Английский

---

In Stallion Spermatozoa, Superoxide Dismutase (Cu–Zn) (SOD1) and the Aldo-Keto-Reductase Family 1 Member b (AKR1B1) Are the Proteins Most Significantly Reduced by Cryopreservation

Journal of Proteome Research, Journal Year: 2021, Volume and Issue: 20(5), P. 2435 - 2446

Published: March 3, 2021

Although cryopreservation is widely used in animal breeding, the technique still suboptimal. The population of spermatozoa surviving procedure experiences changes attributed to alteration their redox regulation. In order expand our knowledge regarding this particular aspect, proteome fresh and frozen thawed aliquots equine was studied identify proteins most severely affected by procedure. If regulation a major factor explaining cryodamage, participating should be principally affected. Using split sample design, 30 ejaculates from 10 different stallions were analyzed as spermatozoa, another aliquot same ejaculate sample. under both conditions using UHPLC-MS/MS bioinformatic analysis conducted discriminant variables between conditions. Data are available through ProteomeXchange Consortium with identifier PXD022236. significantly reduced Aldo-keto reductase family 1 member B (p = 2.2 × 10-17) Superoxide dismutase (Cu-Zn) 4.7 10-14). This first time that SOD1 has been identified discriminating variable analysis, where it one highly seen semen. finding strongly supports theory oxidative stress involved cryodamage suggests control target improve current procedures.

Language: Английский

---

Differences in the proteome of stallion spermatozoa explain stallion-to-stallion variability in sperm quality post-thaw†

Biology of Reproduction, Journal Year: 2021, Volume and Issue: 104(5), P. 1097 - 1113

Published: Jan. 12, 2021

The identification of stallions and or ejaculates that will provide commercially acceptable quality post-thaw before cryopreservation is great interest, avoiding wasting time resources freezing not achieve sufficient to be marketed. Our hypothesis was after bioinformatic analysis, the study stallion sperm proteome can discriminant variables able predict ejaculate. At least three from 10 different were frozen following a split sample design. Half ejaculate analyzed as fresh aliquot other half then frozen-thawed aliquot. Computer-assisted analysis flow cytometry used analyze quality. Detailed proteomic performed on thawed aliquots, identify in samples outcome cryopreservation. Those with fold change > 3, P = 8.2e-04, q 0.074 (equivalent False discovery rate (FDR)) selected, proteins identified good motility post-thaw: F6YTG8, K9K273, A0A3Q2I7V9, F7CE45, F6YU15, F6SKR3. Other also predictors mitochondrial membrane potential viability post-thaw. We concluded approaches are powerful tool improve current biotechnologies.

Language: Английский

---

Seminal Plasma Cytokines Are Predictive of the Outcome of Boar Sperm Preservation

Frontiers in Veterinary Science, Journal Year: 2019, Volume and Issue: 6

Published: Dec. 4, 2019

Background: Boar seminal plasma is rich in cytokines, which could influence the capability of spermatozoa to tolerate preservation. Objectives: To evaluate involvement boar cytokines changes experienced by during their storage, either liquid or frozen state. Materials and Methods: In two separated experiments, semen samples from healthy fertile boars were split aliquots, one centrifuged twice (1,500 ×g for 10 min) harvest plasma, whereas other was commercially extended (3 × 107 sperm/mL) liquid-stored at 17°C 144 h (n = 28, Experiment 1) frozen-thawed using a standard 0.5 mL protocol 27, 2). Sixteen quantified Luminex xMAP®. Sperm attributes (CASA-evaluated total progressive motility; flow cytometry-evaluated sperm viability, production intracellular H2O2 O2•- levels lipid peroxidation viable spermatozoa) evaluated 0, 72, storage (Experiment before freezing 30- 150-min post-thawing Results: Multiple linear regression models, with Bayesian approach variable selection, revealed that anti-inflammatory TGF-β2, TGF-β3, IL-1Ra, IL-4 pro-inflammatory IL-8 IL-18, predicted motility while IFN-γ included models predicting all cryopreserved semen. Conclusion: Specific would contribute modulate structural metabolic shown preservation,

Language: Английский

---

Proteomics in fresh and preserved pig semen: Recent achievements and future challenges

Theriogenology, Journal Year: 2020, Volume and Issue: 150, P. 41 - 47

Published: Jan. 30, 2020

Language: Английский

---

Epididymal and ejaculated sperm differ on their response to the cryopreservation and capacitation processes in mouflon (Ovis musimon)

Scientific Reports, Journal Year: 2019, Volume and Issue: 9(1)

Published: Oct. 30, 2019

Abstract Spermatozoa must undergo the process of capacitation to fertilize egg which involves a cell destabilizing process. Capacitation-like changes such as protein tyrosine phosphorylation (PTP) are associated with cryopreservation. The aim this study was compare cryoresistance and response epididymal ejaculated sperm European mouflon ( Ovis musimon ). Post-thaw parameters were analysed from samples cryopreserved by slow-freezing or ultrarapid-freezing for comparison. Sperm status assessed semiquantification PTP levels, localization kinematic clustering. Epididymal had higher than in both freezing techniques, rendered better results samples. Ejaculated levels and, additionally, showed capacitating (CA) non-capacitating (NCA) conditions while there no effect medium sperm. There tail CA NCA types Kinematic analysis revealed that cluster hyperactivated movement increased incubated whereas observed clusters. In conclusion, freezability lower compared

Language: Английский

---

The proteome of frozen-thawed pig spermatozoa is dependent on the ejaculate fraction source

Scientific Reports, Journal Year: 2019, Volume and Issue: 9(1)

Published: Jan. 24, 2019

Abstract The preservation of sperm functional parameters and fertility post-cryopreservation largely varies in the porcine, a species with fractionated ejaculate. Although intrinsic individual differences have primarily been linked to this variation, protein abundance among frozen-thawed (FT)-spermatozoa are far more relevant. This study, performed two experiments, looked for proteomic quantitative between FT-sperm samples differing post-thaw viability, motility, apoptosis, membrane lipid peroxidation nuclear DNA fragmentation. spermatozoa were either derived from sperm-rich ejaculate fraction (SRF) or entire (Experiment 1) first 10 mL SRF, remaining SRF post-SRF 2). Quantitative analysed using LC-ESI-MS/MS-based SWATH approach. In Experiment 1, FT-spermatozoa showed better than those ejaculate, 26 Sus scrofa proteins relevance showing relative (FC ≥ 1.5) sources. 2, qualitatively post-SRF, 187 three results indicate that proteome cryosurvival.

Language: Английский

---

How does the boar epididymis regulate the emission of fertile spermatozoa?

Animal Reproduction Science, Journal Year: 2021, Volume and Issue: 246, P. 106829 - 106829

Published: Aug. 23, 2021

The epididymis is responsible for peripheral immune tolerance of maturing spermatozoa even though these have xeno-antigens foreign to the male and female system. also produces factors required fertilization serves as a sperm repository until time ejaculation. These reproduction-relevant epididymal functions occur in mesonephros-derived duct-system that composed absorptive secretory epithelial cells with capacity merocrine apocrine secretion proteins, antioxidative- electrolyte/pH-regulating enzymes small, non-coding RNAs (sncRNAs), many stored epididymosomes adhesion long-lasting modifications functions. This paper provides review summary current new knowledge how boar affects quality ejaculate breeding boars. There particular focus on maturation, survival, function role signaling system fertility modulation. Furthermore, aspects related ductus contributions regarding electrolyte control, protein production, release contain sncRNAs are emphasized novel associations male, quiescence during storage cauda epididymis, changes occurring subsequent

Language: Английский

---