Lucas Arjona, Xiomara

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Lucas Arjona, Xiomara

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Lucas Arjona, Xiomara

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Universidad de Murcia. Departamento de Medicina y Cirugía Animal

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Colour and pulsed Doppler Ultrasonographic Study of the canine testis
(Wiley , 2012-07-10) Carrillo Sánchez, J. D.; Soler Laguía, Marta; Lucas Arjona, Xiomara; Agut Giménez, Amalia; Medicina y Cirugía Animal; Facultades de la UMU::Facultad de Veterinaria
Este estudio se llevó a cabo para caracterizar el flujo sanguíneo normal del testículo canino y medir la velocidad sistólica máxima (PSV), la velocidad telediastólica (EDV), el índice de resistencia (RI) y el índice de pulsatilidad (PI) de las arterias testiculares semanalmente durante un período de 6 meses en cinco perros Beagle sanos, así como para evaluar si se producían cambios a lo largo de este tiempo. Los exámenes ecográficos se realizaron con un transductor lineal de 11 MHz. Los vasos testiculares se subdividieron en tres categorías: arterias supratesticulares, arteria marginal y vasos intratesticulares. En las arterias supratesticulares se registraron dos mediciones, en la porción craneal y en la porción en asa (looping). No se observaron diferencias significativas en ninguno de los parámetros estudiados durante los 6 meses que duró el estudio. La porción craneal de la arteria supratesticular mostró un patrón de flujo característico de vaso de alta resistencia, mientras que en la porción en asa de la arteria supratesticular, así como en las arterias marginales e intratesticulares, el flujo presentó un patrón de baja resistencia. Los valores de PSV, RI y PI fueron más elevados en la porción craneal de la arteria supratesticular, seguidos por la porción en asa de la arteria supratesticular, la arteria marginal y los vasos intratesticulares. Las mediciones de EDV fueron mayores en la porción en asa de la arteria supratesticular.
Open Access
Proteomic profiling of porcine seminal extracellular vesicles reveals potential in vivo fertility biomarkers
(Wiley, 2025-07-04) Barranco Cascales, Isabel; Martínez Díaz, Pablo; Parra, Ana; Martínez-Alborcia, María José; Lucas Arjona, Xiomara; Rodríguez-Martínez, Heriberto; Roca, Jordi; Medicina y Cirugía Animal; Facultad de Veterinaria
Background: Predicting male fertility in farm animals remains a challenge. Seminal plasma (SP) contains a high amount of heterogeneous seminal extracellular vesicles (sEVs), believed involved in reproductive processes and maybe key to understanding male fertility. Aims: To identify the sEV proteins that are differentially expressed between more and less fertile boars and that could be candidates for fertility biomarkers in boars used in artificial insemination (AI) programs. Materials and methods: Small (S) and large (L) sEV subsets from SP samples of AI boars with differences in fertility: high (H) or low (L) farrowing rate (FR) and large (L) or small (S) litter size (LS). The S- and L-sEV subsets were isolated by size exclusion chromatography and characterized according to the Minimal Information for Studies of Extracellular Vesicles (MISEV2023) guidelines. Proteomic analyses (three biological replicates per fertility group and sEV subset) were performed using a Bruker timsTOF fleX™ instrument with data-independent acquisition parallel accumulation-serial fragmentation (diaPASEF) technology. Results: A total of 470 and 726 proteins were quantified in S-sEVs and 1801 and 1834 proteins in L-sEVs from FR and LS boars, respectively. Differentially expressed sEV proteins (log2fold change ≥±1, p ≤ 0.05 and effect size d of Cohen >2.0) were found between the fertility groups: seven in S-sEVs and 52 in L-sEVs between H-FR and L-FR boars, and 47 in S-sEVs and 52 in L-sEVs between L-LS and S-LS boars. Many of these differentially expressed sEV proteins are involved in reproductive processes, particularly in sperm function and sperm-zona pellucida binding, but also in embryo development and implantation. Conclusions: The sEV proteome differs between more and less fertile boars, with many of the differentially expressed proteins known as involved in reproductive processes. This would suggest that sEVs may be involved in male fertility and that some of the differentially expressed sEV proteins could be potential fertility markers for AI boars.
Open Access
Protective role of extracellular vesicles against oxidative DNA damage
(BioMed Central, 2025-03-13) Ribas Maynou, Jordi; Parra, Ana; Martínez Díaz, Pablo; Peres Rubio, Camila; Lucas Arjona, Xiomara; Yeste, Marc; Roca, Jordi; Barranco Cascales, Isabel; Medicina y Cirugía Animal; Facultad de Veterinaria
Background: Oxidative stress, a source of genotoxic damage, is often the underlying mechanism in many functional cell disorders. Extracellular vesicles (EVs) have been shown to be key regulators of cellular processes and may be involved in maintaining cellular redox balance. Herein, we aimed to develop a method to assess the effects of EVs on DNA oxidation using porcine seminal plasma extracellular vesicles (sEVs). Results: The technique was set using a cell-free plasmid DNA to avoid the bias generated by the uptake of sEVs by sperm cells, employing increasing concentrations of hydrogen peroxide (H2O2) that generate DNA single-strand breaks (SSBs). Because SSBs contain a free 3'-OH end that allow the extension through quantitative PCR, such extension -and therefore the SYBR intensity- showed to be proportional to the amount of SSB. In the next stage, H2O2 was co-incubated with two size-differentiated subpopulations (small and large) of permeabilized and non-permeabilized sEVs to assess whether the intravesicular content (IC) or the surface of sEVs protects the DNA from oxidative damage. Results obtained showed that the surface of small sEVs reduced the incidence of DNA SSBs (P < 0.05), whereas that of large sEVs had no impact on the generation of SSBs (P > 0.05). The IC showed no protective effect against DNA oxidation (P > 0.05). Conclusions: Our results suggest that the surface of small sEVs, including the peripheral corona layer, may exert a protective function against alterations that are originated by oxidative mechanisms. In addition, our in vitro study opens path to detect, localize and quantify the protective effects against oxidation of extracellular vesicles derived from different fluids, elucidating their function in physiopathological states.
Open Access
RNA profiles differ between small and large extracellular vesicle subsets isolated from porcine seminal plasma
(BioMed Central, 2024-12-27) Barranco Cascales, Isabel; Almiñana, Carmen; Parra, Ana; Martínez Díaz, Pablo; Lucas Arjona, Xiomara; Bauersachs, Stefan; Roca Aleu, Jorge; Medicina y Cirugía Animal
Background: Extracellular vesicles (EVs) are essential for cell-to-cell communication because they transport functionally active molecules, including proteins, RNA, and lipids, from secretory cells to nearby or distant target cells. Seminal plasma contains a large number of EVs (sEVs) that are phenotypically heterogeneous. The aim of the present study was to identify the RNA species contained in two subsets of porcine sEVs of different sizes, namely small sEVs (S-sEVs) and large sEVs (L-sEVs). The two subsets of sEVs were isolated from 54 seminal plasma samples by a method combining serial centrifugations, size exclusion chromatography, and ultrafiltration. The sEVs were characterized using an orthogonal approach. Analysis of RNA content and quantification were performed using RNA-seq analysis. Results: The two subsets of sEVs had different size distributions (P < 0.001). They also showed differences in concentration, morphology, and specific protein markers (P < 0.05). A total of 735 RNAs were identified and quantified, which included: (1) mRNAs, rRNAs, snoRNAs, snRNAs, tRNAs, other ncRNAs (termed as "all RNAs"), (2) miRNAs and (3) piRNAs. The distribution pattern of these RNA classes differed between S-sEVs and L-sEVs (P < 0.05). More than half of "all RNAs", miRNAs and piRNAs were found to be differentially abundant between S- and L-sEVs (FDR < 0.1%). Among the differentially abundant RNAs, "all RNAs" were more abundant in L- than in S-sEVs, whereas the most of the miRNAs were more abundant in S- than in L-sEVs. Differentially abundant piRNAs were equally distributed between S- and L-sEVs. Some of the all RNAs and miRNAs found to be differentially abundant between S- and L-sEVs were associated with sperm quality and functionality and male fertility success. Conclusions: Small and large sEVs isolated from porcine seminal plasma show quantitative differences in RNA content. These differences would suggest that each sEV subtype exerts different functional activities in the targeted cells, namely spermatozoa and functional cells of the female reproductive tract.
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.