DigitalUM :: Browsing by Subject "Spermatozoa"

Browsing by Subject "Spermatozoa"

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Open Access
A fine structural review on the spermatozoa of Cyprinidae with attention to their phylogenetic implications
(Murcia : F. Hernández, 2009) Fürböck, Sonja; Lahnsteiner, Franz; Patzner, Robert A.
The fine structural organization and dimensions of spermatozoa from species of 4 subfamilies of the Cyprinidae (Barbus barbus, Carassius carassius, Cyprinus carpio carpio, Cyprinus carpio haematopterus, Abramis brama, Alburnoides bipunctatus, Alburnus alburnus, Chalcalburnus chalcoides mento, Chondrostoma nasus, Hypophthalmichthys molitrix, Leuciscus cephalus, Phoxinus phoxinus, Rutilus rutilus, Rutilus meidingerii, Scardinius erythrophthalmus, Vimba vimba and Ctenopharyngodon idella) are compared with each other as well as with results from other studies. Based on these descriptions it is investigated whether sperm structure reveals correlations with the existing systematics and if it could be a useful taxonomical parameter. The scatter plots based on the discriminate analysis and the neighbour-joining trees based on a Mahalanobis distance matrix reveal that sperm organization is related with systematics in many aspects. However, in some cases there are also clear differences between relations found on the basis of sperm morphology and between the systematic relations.
Open Access
Aquaporins are essential to maintain motility and membrane lipid architecture during mammalian sperm capacitation
(Frontiers Media, 2021-09-01) Delgado-Bermúdez, Ariadna; Recuero, Sandra; Llavanera, Marc; Mateo-Otero, Yentel; Sandu, Andra; Barranco Cascales, Isabel; Ribas-Maynou, Jordi; Yeste, Marc; Medicina y Cirugía Animal; Facultad de Veterinaria
Aquaporins are a family of ubiquitous transmembrane proteins that allow the transport of water and small molecules across the cell plasma membrane. The different members of this family present a characteristic distribution across different cell types, which is species-specific. In mammalian sperm, different AQPs, including AQP3, AQP7, and AQP11, have been identified; their main roles are related to osmoadaptation and sperm motility activation after ejaculation. Capacitation, which is a post-ejaculatory process that sperm must undergo to achieve fertilizing ability, is triggered by pH changes and different extracellular ions that are present in the female reproductive tract. Considering the function of AQPs and their influence on pH through the regulation of water flow, this study aimed to elucidate the potential role of different AQPs during in vitro sperm capacitation using three different transition metal compounds as AQP inhibitors. Cooper sulfate, a specific inhibitor of AQP3, caused a drastic increase in peroxide intracellular levels compared to the control. Mercury chloride, an unspecific inhibitor of all AQPs except AQP7 produced an increase in membrane lipid disorder and led to a decrease in sperm motility and kinetics parameters. Finally, the addition of silver sulfadiazine, an unspecific inhibitor of all AQPs, generated the same effects than mercury chloride, decreased the intracellular pH and altered tyrosine phosphorylation levels after the induction of the acrosome reaction. In the light of the aforementioned, (a) the permeability of AQP3 to peroxides does not seem to be crucial for sperm capacitation and acrosome reaction; (b) AQPs have a key role in preserving sperm motility during that process; and (c) AQPs as a whole seem to contribute to the maintenance of lipid membrane architecture during capacitation and may be related to the intracellular signaling pathways involved in the acrosome reaction. Hence, further research aimed to elucidate the mechanisms underlying the involvement of AQPs in mammalian sperm capacitation and acrosome reaction is warranted.
Open Access
Artificial insemination of all ejaculated sperm fractions accelerates embryo development and increases the uterine vascularity in the pig
(Elsevier, 2024-04-15) Toledo Guardiola, Santa María; Luongo, Chiara; García Vázquez, Francisco A.; Soriano Úbeda, C.; Matás, C.; Párraga Ros, Ester; Seva Alcaraz, Juan; Anatomía y Anatomía Patológica Comparadas
The semen of boar is characterized by ejaculation in well-differentiated fractions with specific concentration, composition, and volume. The ‘sperm-rich fraction' (SRF), the most concentrated seminal fraction, is habitually collected in insemination centers to make artificial insemination (AI) doses. The absence of the other fractions in AI doses could alter the uterine reaction to AI and not trigger essential responses that could maximize fertility. Thus, there is an urge to ascertain the impact of different ejaculate fractions on the uterus after AI to optimize the semen doses. This work analyzed specific parameters related to fertility in pregnant artificially inseminated sows (n = 15) with ac-cumulative fractions of the semen of boars (n = 6): F1, composed of the sperm-rich fraction (SRF); F2, composed of F1 plus the intermediate fraction; F3, composed of F2 plus the post-SRF. Non-inseminated sows (n = 5) were included as control (C). The different types of seminal dose did not affect the number of ovulated follicles (CL; corpora lutea, p > 0.05) but did affect the embryo development (p < 0.05). The proportion of embryos in morula stages was significantly higher in AI-F1 sows (84.4%, p < 0.05). Morulas and blastocysts were balanced in AI-F2 or AI-F3 (p > 0.05). Independently of the type of seminal dose (F1, F2, or F3), we observed by immunohistochemistry that AI significantly increased uterine vascularization, although with some anatomical differences. The cranial region of the uterine horns was significantly more vascularized in AI-F1 or AI-F2 sows (26.7 ± 2.3 and 28.6 ± 2.0%, respectively), and AI-F3 showed significantly less vascularization at that point (17.8 ± 1.6%, p < 0.05). To summarize, the synergistic effect of all ejaculate fractions accelerates embryo development, at least during the preimplantation period, and increases the uterine reaction to AI in certain parts of the uterus.
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Boar semen proteomics and sperm preservation
(Elsevier, 2019-06-02) Parrilla, Inmaculada; Pérez Patiño, Cristina; Li, J.; Barranco Cascales, Isabel; Padilla, L.; Rodriguez-Martínez, Heriberto; Martínez, E. A.; Roca J.; Medicina y Cirugía Animal; Facultad de Veterinaria
Recently numerous proteomic approaches have been undertaken to identify sperm and seminal plasma (SP) proteins that can be used as potential biomarkers for sperm function, including fertilization ability. This review aims firstly to briefly introduce the proteomic technologies and workflows that can be successfully applied for sperm and SP proteomic analysis. Secondly, we summarize the current knowledge about boar SP and the sperm proteome, focusing mainly on its relevance to sperm preservation procedures (liquid storage or cryopreservation) and their outcomes in terms of sperm function and fertility.
Open Access
Cryopreservation differentially alters the proteome of epididymal and ejaculated pig spermatozoa
(2019-04-11) Perez-Patiño, Cristina; Barranco, Cascales; Li, Junwei; Padilla, Lorena; Martínez, Emilio A; Rodriguez-Martinez, Heriberto; Roca, Jordi; Parrilla, Inmaculada; Medicina y Cirugía Animal
Cryopreservation induces differential remodeling of the proteome in mammalian spermatozoa. How these proteome changes relate to the loss of sperm function during cryopreservation remains unsolved. The present study aimed to clarify this issue evaluating differential changes in the proteome of fresh and frozen-thawed pig spermatozoa retrieved from the cauda epididymis and the ejaculate of the same boars, with clear differences in cryotolerance. Spermatozoa were collected from 10 healthy, sexually mature, and fertile boars, and cryopreserved using a standard 0.5 mL-straw protocol. Total and progressive motility, viability, and mitochondria membrane potential were higher and membrane fluidity and reactive oxygen species generation lower in frozen-thawed (FT) epididymal than ejaculated spermatozoa. Quantitative proteomics of fresh and FT spermatozoa were analyzed using a LC-ESI-MS/MS-based Sequential Window Acquisition of All Theoretical Spectra approach. Cryopreservation quantitatively altered more proteins in ejaculated than cauda epididymal spermatozoa. Differential protein-protein networks highlighted a set of proteins quantitatively altered in ejaculated spermatozoa, directly involved in mitochondrial functionality which would explain why ejaculated spermatozoa deteriorate during cryopreservation.
Open Access
Effect of AQP inhibition on boar sperm cryotolerance depends on the intrinsic freezability of the ejaculate
(MDPI, 2019-12-11) Delgado Bermúdez, Ariadna; LLavanera, Marc; Recuero, Sandra; Mateo Otero, Yentel; Bonet, Sergi; Barranco Cascales, Isabel; Fernández-Fuertes, Beatriz; Yeste, Marc; Medicina y Cirugía Animal; Facultad de Veterinaria
Aquaporins (AQPs) are transmembrane channels with permeability to water and small solutes that can be classified according to their structure and permeability into orthodox AQPs, aquaglyceroporins (GLPs), and superAQPs. In boar spermatozoa, AQPs are related to osmoregulation and play a critical role in maturation and motility activation. In addition, their levels differ between ejaculates with good and poor cryotolerance (GFE and PFE, respectively). The aim of this work was to elucidate whether the involvement of AQPs in the sperm response to cryopreservation relies on the intrinsic freezability of the ejaculate. With this purpose, two different molecules: phloretin (PHL) and 1,3-propanediol (PDO), were used to inhibit sperm AQPs in GFE and PFE. Boar sperm samples were treated with three different concentrations of each inhibitor prior to cryopreservation, and sperm quality and functionality parameters were evaluated in fresh samples and after 30 and 240 min of thawing. Ejaculates were classified as GFE or PFE, according to their post-thaw sperm viability and motility. While the presence of PHL caused a decrease in sperm quality and function compared to the control, samples treated with PDO exhibited better quality and function parameters than the control. In addition, the effects of both inhibitors were more apparent in GFE than in PFE. In conclusion, AQP inhibition has more notable consequences in GFE than in PFE, which can be related to the difference in relative levels of AQPs between these two groups of samples.
Open Access
Effect of cumulus cell removal and sperm pre‐incubation with progesterone on in vitro fertilization of equine gametes in the presence of oviductal fluid or cells
(2019-05-30) Douet, C; Reigner, F; Goudet, G; Moros Nicolás, Carla; Biología Celular e Histología
In spite of many attempts to establish an in vitro fertilization (IVF) technique in the equine, no efficient conventional IVF technique is available. The presence of oviductal fluid or oviductal cells during IVF help to improve embryo production in vitro but is not sufficient to reach high fertilization rates. Thus, our aim was to perform equine IVF either after sperm preincubation with oviductal fluid or in the presence of oviductal cells, and to evaluate the effect of cumulus removal from the oocyte or sperm preincubation with progesterone. In experiment 1 and 2, IVF was performed in the presence of porcine oviduct epithelial cells. The removal of cumulus cells from equine oocytes after in vitro maturation tended to increase the percentage of fertilization when fresh sperm was used (1/33 vs 4/31, p > 0.05) but had no effect when frozen sperm was used (1/32 vs 1/32). Equine sperm preincubation with progesterone did not significantly influence the fertilization rate when fresh or frozen sperm was used (2/14 vs 2/18 for fresh, 1/29 vs 1/25 for frozen). In experiment 3 and 4, IVF was performed after preincubation of sperm with porcine oviductal fluid. The removal of cumulus cells tented to increase the percentage of fertilization when fresh sperm was used (1/24 vs 3/26, p>0.05). Sperm preincubation with progesterone did not significantly influence the fertilization rate when fresh or frozen sperm was used (2/39 vs 2/36 for fresh, 2/37 vs 1/46 for frozen), but two 3-4 cells stage zygotes were obtained with fresh sperm preincubated with progesterone. This is an encouraging result for the setting up of an efficient IVF procedure in equine.
Open Access
Seminal extracellular vesicles influence porcine spermatozoa physiology by modulating key functional parameters
(Elsevier, 2025-09-27) Parra, Ana; Martín-Cano, Francisco E.; Martínez Díaz, Pablo; Panales, Patricia; Lucas Arjona, Xiomara; Roca, Jordi; Barranco Cascales, Isabel; Peña, Fernando J.; Medicina y Cirugía Animal; Facultad de Veterinaria
Seminal plasma (SP) contains a heterogeneous population of extracellular vesicles (EVs) recognized as key modulators of sperm function. However, the specific functional roles of each seminal EV (sEV) subset remain poorly understood. This study aimed to evaluate the interaction of two sized sEV subsets (small [S-sEVs] and large [L-sEVs]) with pig liquid-stored spermatozoa under different pH conditions and their effect on specific sperm functional parameters. Seminal EV subsets were isolated from SP samples using size exclusion chromatography and characterized following the MISEV2023 guidelines. Semen samples were incubated with each sEV subset or without sEVs (control) for 6 h at 37 ºC, 100 % humidity and 5 % CO₂ under different pH conditions (6.5, 7.0, or 7.5). Sperm functional parameters were assessed by flow cytometry (Cytoflex®S and LX, Beckman Coulter), under capacitating and non-capacitating conditions. Confocal microscopy revealed that both sEV subsets bound to and were internalized by spermatozoa as early as 30 min after incubation, regardless of pH. Flow cytometry revealed that both sEVs decreased reactive oxygen species production (P ≤ 0.0001), mitochondrial membrane potential (P ≤ 0.0001) and mitochondrial O₂•⁻ levels (P ≤ 0.01) and increased apoptosis (active caspase-3) in viable spermatozoa (P ≤ 0.0001). However, the influence of sEV on acrosome integrity in viable sperm was time- and condition-dependent (P ≤ 0.05). This study showed that both S- and L-sEVs interact with porcine spermatozoa across a range of physiological pH conditions. This interaction is reflected by decreased oxidative stress and mitochondrial activity, as well as increased apoptosis in spermatozoa.
Open Access
Seminal plasma cytokines are predictive of the outcome of boar sperm preservation
(2019-12-04) Barranco, Isabel; Padilla, Lorena; Perez-Patiño, Cristina; Vazquez, Juan M; Martinez, Emilio A; Rodriguez-Martinez, Heriberto; Roca, Jordi; Parrilla, Inmaculada; Medicina y Cirugía Animal
Background: Boar seminal plasma is rich in cytokines, which could influence the capability of spermatozoa to tolerate preservation. Objectives: To evaluate the involvement of boar seminal plasma cytokines in the changes experienced by boar spermatozoa during their storage, either in liquid or frozen state. Materials and Methods: In two separated experiments, semen samples from healthy and fertile boars were split in two aliquots, one centrifuged twice (1,500 ×g for 10 min) to harvest seminal plasma, whereas the other was either commercially extended (3 × 107 sperm/mL) and liquid-stored at 17°C during 144 h (n = 28, Experiment 1) or frozen-thawed using a standard 0.5 mL protocol (n = 27, Experiment 2). Sixteen cytokines were quantified using Luminex xMAP®. Sperm attributes (CASA-evaluated total and progressive motility; flow cytometry-evaluated sperm viability, production of intracellular H2O2 and O∙−2 and levels of lipid peroxidation in viable spermatozoa) were evaluated either at 0, 72, or 144 h of liquid storage (Experiment 1) or before freezing and at 30- and 150-min post-thawing (Experiment 2). Results: Multiple linear regression models, with Bayesian approach for variable selection, revealed that the anti-inflammatory TGF-β2, TGF-β3, IL-1Ra, and IL-4 and the pro-inflammatory IL-8 and IL-18, predicted changes in sperm motility for liquid-stored semen while the anti-inflammatory IFN-γ was included in the models predicting changes in all sperm attributes for cryopreserved semen. Conclusion: Specific boar seminal plasma cytokines would contribute to modulate the structural and metabolic changes shown by spermatozoa during preservation, either in liquid or frozen state.
Open Access
Sperm methylome profiling can discern fertility levels in the porcine biomedical model
(MDPI, 2021-03-06) Pértille, Fabio; Álvarez Rodríguez, Manuel; Da Silva, Arthur Nery; Barranco, Isabel; Roca, Jordi; Guerrero Bosagna, Carlos; Rodríguez Martínez, Heriberto; Medicina y Cirugía Animal
A combined Genotyping By Sequencing (GBS) and methylated DNA immunoprecipitation (MeDIP) protocol was used to identify-in parallel-genetic variation (Genomic-Wide Association Studies (GWAS) and epigenetic differences of Differentially Methylated Regions (DMR) in the genome of spermatozoa from the porcine animal model. Breeding boars with good semen quality (n = 11) and specific and well-documented differences in fertility (farrowing rate, FR) and prolificacy (litter size, LS) (n = 7) in artificial insemination programs, using combined FR and LS, were categorized as High Fertile (HF, n = 4) or Low Fertile (LF, n = 3), and boars with Unknown Fertility (UF, n = 4) were tested for eventual epigenetical similarity with those fertility-proven. We identified 165,944 Single Nucleotide Polymorphisms (SNPs) that explained 14-15% of variance among selection lines. Between HF and LF individuals (n = 7, 4 HF and 3 LF), we identified 169 SNPs with p ≤ 0.00015, which explained 58% of the variance. For the epigenetic analyses, we considered fertility and period of ejaculate collection (late-summer and mid-autumn). Approximately three times more DMRs were observed in HF than in LF boars across these periods. Interestingly, UF boars were clearly clustered with one of the other HF or LF groups. The highest differences in DMRs between HF and LF experimental groups across the pig genome were located in the chr 3, 9, 13, and 16, with most DMRs being hypermethylated in LF boars. In both HF and LF boars, DMRs were mostly hypermethylated in late-summer compared to mid-autumn. Three overlaps were detected between SNPs (p ≤ 0.0005, n = 1318) and CpG sites within DMRs. In conclusion, fertility levels in breeding males including FR and LS can be discerned using methylome analyses. The findings in this biomedical animal model ought to be applied besides sire selection for andrological diagnosis of idiopathic sub/infertility.
Open Access
Sperm proteome after interaction with reproductive fluids in porcine: from the ejaculation to the fertilization site
(MDPI, 2020-08-22) Luongo, Chiara; Gónzalez-Brusi, Leopoldo; García-Vázquez, Francisco Alberto; Cots Rodríguez, Paula; Izquierdo Rico, María José; Avilés Sánchez, Manuel; Biología Celular e Histología
Ejaculated sperm are exposed to different environments before encountering the oocyte. However, how the sperm proteome changes during this transit remains unsolved. This study aimed to identify proteomic changes in boar sperm after incubation with male (seminal plasma, SP) and/or female (uterine fluid, UF; and oviductal fluid, OF) reproductive fluids. The following experimental groups were analyzed: (1) SP: sperm + 20% SP; 2) UF: sperm + 20% UF; 3) OF: sperm + 20% OF; 4) SP + UF: sperm + 20% SP + 20% UF; and (5) SP+OF: sperm + 20% SP + 20% OF. The proteome analysis, performed by HPLC-MS/MS, allowed the identification of 265 proteins. A total of 69 proteins were detected in the UF, SP, and SP + UF groups, and 102 proteins in the OF, SP, and SP + OF groups. Our results showed a higher number of proteins when sperm were incubated with only one fluid than when they were co-incubated with two fluids. Additionally, the number of sperm-interacting proteins from the UF group was lower than the OF group. In conclusion, the interaction of sperm with reproductive fluids alters its proteome. The description of sperm-interacting proteins in porcine species after co-incubation with male and/or female reproductive fluids may be useful to understand sperm transport, selection, capacitation, or fertilization phenomena.
Open Access
The oxytocin receptor in spermatozoa may originate from both spermatogenesis and epididymal maturation, and regulates capacitation
(Wiley, 2025-09-27) Garriga, Ferran; Ahmad, Adeel; Padilla, Lorena; Maside, Carolina; Bonet, Sergi; Barranco Cascales, Isabel; Roca, Jordi; Pastor García, Luis Miguel; Yeste, Marc; Martínez Hernández, Jesús; Medicina y Cirugía Animal; Facultad de Veterinaria
Background: The oxytocin receptor (OR) is a G-protein-coupled receptor recently identified in human spermatozoa, whose origin and role in sperm physiology remain unknown. Objectives: In this study, using the pig as a model, we examine the presence of the OR in ejaculated spermatozoa through immunofluorescence and immunoblotting, and investigate the receptor's origin in the male gamete via immunohistochemistry in testicular and epididymal tissues. Additionally, we assess the involvement of the OR in in vitro capacitation and the acrosome reaction by utilizing physiological concentrations of agonists (oxytocin and carbetocin) and an antagonist (L-371,257). Results: The results indicate that, in addition to the expected presence in ejaculated spermatozoa, the OR is expressed during spermatogenesis. Besides, this receptor is found in Leydig and Sertoli cells, as well as in the principal, basal, and apical cells of the epididymis. Furthermore, our data suggest that, during epididymal maturation, the OR could be incorporated in spermatozoa via extracellular vesicles within the apical blebs. The OR is involved in sperm capacitation, as the combination of the antagonist (L-371,257) and the agonist (carbetocin) increases intracellular calcium levels and membrane lipid disorders, which are known as capacitation markers. Conclusions: The presence of the OR in mammalian spermatozoa could originate from both spermatogenesis and epididymal maturation. Moreover, in the male gamete, this receptor regulates sperm capacitation by interacting with its ligand in the female reproductive tract.
Open Access
The transcriptome of pig spermatozoa, and its role in fertility
(MDPI, 2020-02-25) Álvarez Rodríguez, Manuel; Martínez, Cristina; Wright, Dominic; Barranco, Isabel; Roca, Jordi; Rodríguez Martínez, Heriberto; Medicina y Cirugía Animal
In the study presented here we identified transcriptomic markers for fertility in the cargo of pig ejaculated spermatozoa using porcine-specific micro-arrays (GeneChip® miRNA 4.0 and GeneChip® Porcine Gene 1.0 ST). We report (i) the relative abundance of the ssc-miR-1285, miR-16, miR-4332, miR-92a, miR-671-5p, miR-4334-5p, miR-425-5p, miR-191, miR-92b-5p and miR-15b miRNAs, and (ii) the presence of 347 up-regulated and 174 down-regulated RNA transcripts in high-fertility breeding boars, based on differences of farrowing rate (FS) and litter size (LS), relative to low-fertility boars in the (Artificial Insemination) AI program. An overrepresentation analysis of the protein class (PANTHER) identified significant fold-increases for C-C chemokine binding (GO:0019957): CCR7, which activates B- and T-lymphocytes, 8-fold increase), XCR1 and CXCR4 (with ubiquitin as a natural ligand, 1.24-fold increase), cytokine receptor activity (GO:0005126): IL23R receptor of the IL23 protein, associated to JAK2 and STAT3, 3.4-fold increase), the TGF-receptor (PC00035) genes ACVR1C and ACVR2B (12-fold increase). Moreover, two micro-RNAs (miR-221 and mir-621) were down- and up-regulated, respectively, in high-fertility males. In conclusion, boars with different fertility performance possess a wide variety of differentially expressed RNA present in spermatozoa that would be attractive targets as non-invasive molecular markers for predicting fertility.
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Total urokinase-type plasminogen activator (uPA) levels in seminal plasma are associated with positive assisted reproductive technology outcomes
(2018-03-23) Martínez Soto, Juan Carlos; Landeras, José; Mollá, Marta; Mondéjar, Irene; Nicolás, María; Fernández-Olmedilla, Laura; Trabalón Martina; Coy Fuster, Pilar; Gadea Mateos, Joaquín; Gadea Mateos, Joaquín; Fisiología; Facultades de la UMU

Browsing by Subject "Spermatozoa"

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