261 results on '"Álvarez Rodríguez, Manuel"'
Search Results
2. Effect of the addition of exogenous progesterone and the progesterone receptor inhibitor (RU 486) on boar cryopreservation semen extenders
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Martín-San Juan, Adrián, Gala, Nerea, Nieto-Cristóbal, Helena, Álvarez-Rodríguez, Manuel, and de Mercado, Eduardo
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- 2024
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3. Cooling rate modifies the location of aquaporin 3 in spermatozoa of sheep and goat
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Pequeño, Belén, Millán de la Blanca, María Gemma, Castaño, Cristina, Toledano-Díaz, Adolfo, Esteso, Milagros Cristina, Alba, Esther, Arrebola, Francisco A., Ungerfeld, Rodolfo, Martínez-Madrid, Belén, Alvarez-Rodriguez, Manuel, Rodriguez-Martinez, Heriberto, and Santiago-Moreno, Julián
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- 2024
- Full Text
- View/download PDF
4. MicroRNA expression in specific segments of the pig periovulatory internal genital tract is differentially regulated by semen or by seminal plasma
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Álvarez-Rodríguez, Manuel, Martinez-Serrano, Cristina A., Gardela, Jaume, Nieto, Helena, de Mercado, Eduardo, and Rodríguez-Martínez, Heriberto
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- 2024
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5. Pre-freezing selection of Holstein bull semen with the BoviPure colloid as double- or single-layer centrifugation improves the post-thawing quality
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Salman, Amer, Fernández-Alegre, Estela, Francisco-Vázquez, Rubén, Domínguez, Juan C., Álvarez-Rodríguez, Manuel, Caamaño, J.Néstor, Martínez-Pastor, Felipe, Gómez-Martín, Rubén, Fernández-Fernández, Alejandro, and Areán-Dablanca, Héctor
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- 2023
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6. Cooling rate modifies the location of aquaporin 3 in spermatozoa of sheep and goat
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Agencia Estatal de Investigación (España), Pequeño, Belén [0000-0003-3962-1982], Millán de la Blanca, María Gemma [0000-0002-2810-8510], Castaño, Cristina [0000-0003-1134-1436], Toledano-Díaz, A. [0000-0001-6679-485X], Esteso, Milagros C. [0000-0002-8963-5736], Arrebola, Francisco A. [0000-0002-5571-7258], Ungerfeld, Rodolfo [0000-0003-4685-2105], Martínez-Madrid, Belén [0000-0001-6852-4597], Álvarez-Rodríguez, Manuel [0000-0003-0120-354X], Rodriguez-Martinez, Heriberto [0000-0002-5194-2124], Santiago Moreno, Julián [0000-0001-5551-8120], Pequeño, Belén, Millán de la Blanca, María Gemma, Castaño, Cristina, Toledano-Díaz, A., Esteso, Milagros C., Alba, Esther, Arrebola, Francisco A., Ungerfeld, Rodolfo, Martínez-Madrid, Belén, Álvarez-Rodríguez, Manuel, Rodriguez-Martinez, Heriberto, Santiago Moreno, Julián, Agencia Estatal de Investigación (España), Pequeño, Belén [0000-0003-3962-1982], Millán de la Blanca, María Gemma [0000-0002-2810-8510], Castaño, Cristina [0000-0003-1134-1436], Toledano-Díaz, A. [0000-0001-6679-485X], Esteso, Milagros C. [0000-0002-8963-5736], Arrebola, Francisco A. [0000-0002-5571-7258], Ungerfeld, Rodolfo [0000-0003-4685-2105], Martínez-Madrid, Belén [0000-0001-6852-4597], Álvarez-Rodríguez, Manuel [0000-0003-0120-354X], Rodriguez-Martinez, Heriberto [0000-0002-5194-2124], Santiago Moreno, Julián [0000-0001-5551-8120], Pequeño, Belén, Millán de la Blanca, María Gemma, Castaño, Cristina, Toledano-Díaz, A., Esteso, Milagros C., Alba, Esther, Arrebola, Francisco A., Ungerfeld, Rodolfo, Martínez-Madrid, Belén, Álvarez-Rodríguez, Manuel, Rodriguez-Martinez, Heriberto, and Santiago Moreno, Julián
- Abstract
The freeze-thawing process induces osmotic changes that may affect the membrane domain location of aquaporins’ (AQP) in spermatozoa. Recent studies suggest that changes in AQP3 localization allows better sperm osmo-adaptation, improving the cryoresistance. Ultra-rapid freezing is an alternative cryopreservation technique that requires less equipment than conventional freezing, and it is faster, simpler and can be used in the field. This study aimed to determine the influence of freezing-thawing rates (slow (control) vs. ultra-rapid) on AQP3 expression and location in the spermatozoa from small ruminants (sheep and goats) and its relationship with sperm cryo-damage. Spermatozoa were collected from 10 Merino rams and 10 Murciano-Granadina bucks. The presence and distribution of AQP3 were assessed by Western blotting and immunocytochemistry (ICC), employing a commercial rabbit polyclonal antibody. Sperm motility was CASA system-analyzed, and membrane and acrosome integrity assessed by fluorescence (PI/PNA-FITC). Western blotting did not detect a significant effect of freezing-thawing rate on the amount of AQP3 while ICC found freezing-thawing rate affecting AQP3 location (P<0.05). In both species, the percentages of spermatozoa showing AQP3 in the post-acrosome region, mid-piece, and principal piece of the tail were greater in samples cryopreserved by slow freezing-thawing (control) than ultra-rapid freezing-thawing rates (P<0.05). Spermatozoa cryopreserved using ultra-rapid freezing-thawing showed decrease motility, plasma membrane, and acrosome integrity (P<0.05), which might be related, at least in part, to a lower expression of AQP3. In conclusion, the cooling rate modifies the location of AQP3 in spermatozoa of sheep and goat, which might be associated with sperm cryosurvival.
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- 2024
7. Bicarbonate and BSA increase the capacitation pattern and acrosomal exocytosis in boar sperm after 120 min of incubation
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Agencia Estatal de Investigación (España), Ministerio de Ciencia e Innovación (España), European Commission, Consejo Superior de Investigaciones Científicas (España), Peris-Frau, Patricia [0000-0001-6266-0635], Nieto-Cristóbal, Helena [0000-0003-1192-9467], Santiago Moreno, Julián [0000-0001-5551-8120], de Mercado, Eduardo [0000-0003-0321-2431], Álvarez-Rodríguez, Manuel [000-0003-0120-354X], Dudkiewicz, Sylwia, Peris-Frau, Patricia, Nieto-Cristóbal, Helena, Santiago Moreno, Julián, de Mercado, Eduardo, Álvarez-Rodríguez, Manuel, Agencia Estatal de Investigación (España), Ministerio de Ciencia e Innovación (España), European Commission, Consejo Superior de Investigaciones Científicas (España), Peris-Frau, Patricia [0000-0001-6266-0635], Nieto-Cristóbal, Helena [0000-0003-1192-9467], Santiago Moreno, Julián [0000-0001-5551-8120], de Mercado, Eduardo [0000-0003-0321-2431], Álvarez-Rodríguez, Manuel [000-0003-0120-354X], Dudkiewicz, Sylwia, Peris-Frau, Patricia, Nieto-Cristóbal, Helena, Santiago Moreno, Julián, de Mercado, Eduardo, and Álvarez-Rodríguez, Manuel
- Abstract
Sperm capacitation is a crucial step towards the acquisition of fertilizing capacity. Despite the attempts to mimic the in vivo situation, there is still a lack of standardization in vitro techniques. Bicarbonate and serum albumin (BSA) are routinely used, although controversial results are reported regarding the optimal concentration of each compound. In addition, whether caffeine is needed on in vitro capacitation media in boar sperm remains to be elucidated. Here, 18 boar commercial artificial insemination doses were used to test different concentrations of bicarbonate (19, 37 or 56 mM) in experiment 1, BSA (1.5, 3, 4.5 mg/mL) in experiment 2 and the presence or absence of caffeine (5.15 mM) experiment 3. We analysed at 0, 30 and 120 min of incubation at 38.5°C, 5% CO2 : Total motility (TMOT), membrane integrity (VIAB), acrosomal exocytosis (rAcro; H33342/PI/PNA), capacitation status (chlortetracycline staining CTC) and mitochondrial membrane potential (JC-1). The higher concentrations of bicarbonate (37 and 56 mM) decreased TM and VIAB (p < .01) but increased rAcro (p < .01) after 120 min of incubation compared to the fresh control. In contrast, only the BSA concentration of 3 mg/mL reduced the VIAB at 120 min, but all the concentrations tested increased the average of JC-1 and decreased TM (p < .01) throughout incubation compared to the fresh control. Finally, in experiment 3, when boar sperm were incubated in the capacitating media with bicarbonate, BSA and with or without caffeine, the capacitated pattern measured by the CTC technique and rAcro increased after 120 min of incubation (p < .01) compared to fresh control, either in the presence or in the absence of caffeine. In summary, our results suggested that the combination of capacitating components, like bicarbonate and BSA, contributed to increasing the proportion of capacitated boar spermatozoa, mitochondrial membrane potential as well as acrosomal exocytosis. However, caffeine did not significantly infl
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- 2024
8. Reproductive physiology of the boar: What defines the potential fertility of an ejaculate?
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Ministerio de Ciencia e Innovación (España), Agencia Estatal de Investigación (España), European Commission, Fundación Séneca, Rodriguez-Martinez, Heriberto [0000-0002-5194-2124], Martinez, Cristina A. [0000-0001-6811-0191], Álvarez-Rodríguez, Manuel [0000-0003-0120-354X], Martinez, Emilio A. [0000-0003-1260-9721], Roca, Jordi [0000-0002-4213-3093], Rodriguez-Martinez, Heriberto, Martinez, Cristina A., Álvarez-Rodríguez, Manuel, Martinez, Emilio A., Roca, Jordi, Ministerio de Ciencia e Innovación (España), Agencia Estatal de Investigación (España), European Commission, Fundación Séneca, Rodriguez-Martinez, Heriberto [0000-0002-5194-2124], Martinez, Cristina A. [0000-0001-6811-0191], Álvarez-Rodríguez, Manuel [0000-0003-0120-354X], Martinez, Emilio A. [0000-0003-1260-9721], Roca, Jordi [0000-0002-4213-3093], Rodriguez-Martinez, Heriberto, Martinez, Cristina A., Álvarez-Rodríguez, Manuel, Martinez, Emilio A., and Roca, Jordi
- Abstract
Despite decades of research and handling of semen for use in artificial insemination (AI) and other assisted reproductive technologies, 5-10% of selected boar sires are still considered sub-fertile, escaping current assessment methods for sperm quality and resilience to preservation. As end-product, the ejaculate (emitted spermatozoa sequentially exposed to the composite seminal plasma, the SP) ought to define the homeostasis of the testes, the epididymis, and the accessory sexual glands. Yet, linking findings in the ejaculate to sperm production biology and fertility is suboptimal. The present essay critically reviews how the ejaculate of a fertile boar can help us to diagnose both reproductive health and resilience to semen handling, focusing on methods -available and under development- to identify suitable biomarkers for cryotolerance and fertility. Bulk SP, semen proteins and microRNAs (miRNAs) have, albeit linked to sperm function and fertility after AI, failed to enhance reproductive outcomes at commercial level, perhaps for just being components of a complex functional pathway. Hence, focus is now on the interaction sperm-SP, comparing in vivo with ex vivo, and regarding nano-sized lipid bilayer seminal extracellular vesicles (sEVs) as priority. sEVs transport fragile molecules (lipids, proteins, nucleic acids) which, shielded from degradation, mediate cell-to-cell communication with spermatozoa and the female internal genital tract. Such interaction modulates essential reproductive processes, from sperm homeostasis to immunological female tolerance. sEVs can be harvested, characterized, stored, and manipulated, e.g. can be used for andrological diagnosis, selection of breeders, and alternatively be used as additives to improve cryosurvival and fertility.
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- 2024
9. MicroRNA expression in specific segments of the pig periovulatory internal genital tract is differentially regulated by semen or by seminal plasma
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Agencia Estatal de Investigación (España), European Commission, Álvarez-Rodríguez, Manuel [0000-0003-0120-354X], Martinez, Cristina A. [0000-0001-6811-0191], Gardela, Jaume [0000-0001-7524-2088], Nieto-Cristóbal, Helena [0000-0003-1192-9467], de Mercado, Eduardo [0000-0003-0321-2431], Rodriguez-Martinez, Heriberto [0000-0002-5194-2124], Álvarez-Rodríguez, Manuel, Martinez, Cristina A., Gardela, Jaume, Nieto-Cristóbal, Helena, de Mercado, Eduardo, Rodriguez-Martinez, Heriberto, Agencia Estatal de Investigación (España), European Commission, Álvarez-Rodríguez, Manuel [0000-0003-0120-354X], Martinez, Cristina A. [0000-0001-6811-0191], Gardela, Jaume [0000-0001-7524-2088], Nieto-Cristóbal, Helena [0000-0003-1192-9467], de Mercado, Eduardo [0000-0003-0321-2431], Rodriguez-Martinez, Heriberto [0000-0002-5194-2124], Álvarez-Rodríguez, Manuel, Martinez, Cristina A., Gardela, Jaume, Nieto-Cristóbal, Helena, de Mercado, Eduardo, and Rodriguez-Martinez, Heriberto
- Abstract
microRNAs play pivotal roles during mammalian reproduction, including the cross-talk between gametes, embryos and the maternal genital tract. Mating induces changes in the expression of mRNA transcripts in the female, but whether miRNAs are involved remains to be elucidated. In the current study, we mapped 181 miRNAs in the porcine peri-ovulatory female reproductive tract: Cervix (Cvx), distal and proximal uterus (Dist-Ut, Prox-Ut), Utero-tubal-junction (UTJ), isthmus (Isth), ampulla (Amp), and infundibulum (Inf) when exposed to semen (natural mating (NM) or artificial insemination (AI-P1)) or to infusions of sperm-free seminal plasma (SP): the first 10 mL of the sperm rich fraction (SP-P1) or the entire ejaculate (SP-E). Among the most interesting findings, NM decreased mir-671, implicated in uterine development and pregnancy loss prior to embryo implantation, in Cvx, Dist-UT, Prox-UT, Isth, and Inf, while it increased in Amp. NM and SP-E induced the downregulation of miR-let7A-1 (Dist-UT, Prox-UT), a regulator of immunity during pregnancy. miR-34C-1, a regulator of endometrial receptivity gene expression, was increased in Dist-UT, UTJ and Amp (NM), in Prox-UT (AI-P1), and in Amp (SP-P1). miR-296, a modulator of the inflammatory response and apoptosis, was upregulated in the UTJ (all treatments). NM elicited the highest miRNA activity in the sperm reservoir (UTJ), suggesting that key-regulators such as miR-34c or miR-296 may modulate the metabolic processes linked to the adequate preparation for gamete encounter in the oviduct. Our results suggest that SP should be maintained in AI to warrant miRNA regulation within the female genital tract for reproductive success.
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- 2024
10. Location of aquaporins 3, 7 and 10 in frozen-thawed ejaculated and cauda epididymal spermatozoa from the Iberian ibex, mouflon, and chamois
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Pequeño, Belén, Martínez-Madrid, Belén, Castaño, Cristina, Toledano-Díaz, Adolfo, Bóveda, Paula, Esteso, Milagros C., Gómez-Guillamón, Félix, Prieto, Paloma, Marcos-Beltrán, Jaime L., Alvarez-Rodriguez, Manuel, Rodriguez-Martinez, Heriberto, and Santiago-Moreno, Julián
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- 2023
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11. NR3C1 and glucocorticoid-regulatory genes mRNA and protein expression in the endometrium and ampulla during the bovine estrous cycle
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Ruiz-Conca, Mateo, Gardela, Jaume, Olvera-Maneu, Sergi, López-Béjar, Manel, and Álvarez-Rodríguez, Manuel
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- 2022
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12. The mRNA expression of the three major described cold-inducible proteins, including CIRBP, differs in the bovine endometrium and ampulla during the estrous cycle
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Gardela, Jaume, Ruiz-Conca, Mateo, Olvera-Maneu, Sergi, López-Béjar, Manel, and Álvarez-Rodríguez, Manuel
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- 2022
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13. How does the boar epididymis regulate the emission of fertile spermatozoa?
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Rodriguez-Martinez, Heriberto, Roca, Jordi, Alvarez-Rodriguez, Manuel, and Martinez-Serrano, Cristina A.
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- 2022
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14. Apoptosis and glucocorticoid-related genes mRNA expression is modulated by coenzyme Q10 supplementation during in vitro maturation and vitrification of bovine oocytes and cumulus cells
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Ruiz-Conca, Mateo, Gardela, Jaume, Mogas, Teresa, López-Béjar, Manel, and Álvarez-Rodríguez, Manuel
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- 2022
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15. Sperm freezability is neither associated with the expression of aquaporin 3 nor sperm head dimensions in dromedary camel (Camelus dromedarius)
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O'Brien, Emma, Malo, Clara, Castaño, Cristina, García-Casado, Pedro, Toledano-Díaz, Adolfo, Martínez-Madrid, Belén, Rodriguez-Martinez, Heriberto, Álvarez-Rodríguez, Manuel, and Santiago-Moreno, Julián
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- 2022
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16. Mild hypothermia and vitrification increase the mRNA expression of cold-inducible proteins in bovine oocytes and cumulus cells
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Gardela, Jaume, Ruiz-Conca, Mateo, García-Sanmartín, Josune, Martínez, Alfredo, Mogas, Teresa, López-Béjar, Manel, and Álvarez-Rodríguez, Manuel
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- 2022
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17. mRNA expression of oxidative-reductive proteins in boars with documented different fertility can identify relevant prognostic biomarkers
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Alvarez-Rodriguez, Manuel, Martinez, Cristina A., Roca, Jordi, and Rodriguez-Martinez, Heriberto
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- 2021
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18. Evaluation of Different Thawing Protocols on Iberian Boar Sperm Preserved for 10 Years at Different Liquid Nitrogen Levels
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Álvarez-Rodríguez, Manuel, primary, Tomás-Almenar, Cristina, additional, Nieto-Cristóbal, Helena, additional, and de Mercado, Eduardo, additional
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- 2024
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19. Application of Flow Cytometry Using Advanced Chromatin Analyses for Assessing Changes in Sperm Structure and DNA Integrity in a Porcine Model
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Lacalle, Estíbaliz, primary, Fernández-Alegre, Estela, additional, Gómez-Giménez, Belén, additional, Álvarez-Rodríguez, Manuel, additional, Martín-Fernández, Beatriz, additional, Soriano-Úbeda, Cristina, additional, and Martínez-Pastor, Felipe, additional
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- 2024
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20. Application of Flow Cytometry Using Advanced Chromatin Analyses for Assessing Changes in Sperm Structure and DNA Integrity in a Porcine Model
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Ministerio de Ciencia, Innovación y Universidades (España), Agencia Estatal de Investigación (España), European Commission, Junta de Castilla y León, Lacalle, E., Fernández-Alegre, Estela, Gómez-Giménez, Belén, Álvarez-Rodríguez, Manuel, Martín-Fernández, B., Soriano-Úbeda, C., Martínez-Pastor, F., Ministerio de Ciencia, Innovación y Universidades (España), Agencia Estatal de Investigación (España), European Commission, Junta de Castilla y León, Lacalle, E., Fernández-Alegre, Estela, Gómez-Giménez, Belén, Álvarez-Rodríguez, Manuel, Martín-Fernández, B., Soriano-Úbeda, C., and Martínez-Pastor, F.
- Abstract
Chromatin status is critical for sperm fertility and reflects spermatogenic success. We tested a multivariate approach for studying pig sperm chromatin structure to capture its complexity with a set of quick and simple techniques, going beyond the usual assessment of DNA damage. Sperm doses from 36 boars (3 ejaculates/boar) were stored at 17 °C and analyzed on days 0 and 11. Analyses were: CASA (motility) and flow cytometry to assess sperm functionality and chromatin structure by SCSA (%DFI, DNA fragmentation; %HDS, chromatin maturity), monobromobimane (mBBr, tiol status/disulfide bridges between protamines), chromomycin A3 (CMA3, protamination), and 8-hydroxy-2′-deoxyguanosine (8-oxo-dG, DNA oxidative damage). Data were analyzed using linear models for the effects of boar and storage, correlations, and multivariate analysis as hierarchical clustering and principal component analysis (PCA). Storage reduced sperm quality parameters, mainly motility, with no critical oxidative stress increases, while chromatin status worsened slightly (%DFI and 8-oxo-dG increased while mBBr MFI—median fluorescence intensity—and disulfide bridge levels decreased). Boar significantly affected most chromatin variables except CMA3; storage also affected most variables except %HDS. At day 0, sperm chromatin variables clustered closely, except for CMA3, and %HDS and 8-oxo-dG correlated with many variables (notably, mBBr). After storage, the relation between %HDS and 8-oxo-dG remained, but correlations among other variables disappeared, and mBBr variables clustered separately. The PCA suggested a considerable influence of mBBr on sample variance, especially regarding storage, with SCSA and 8-oxo-dG affecting between-sample variability. Overall, CMA3 was the least informative, in contrast with results in other species. The combination of DNA fragmentation, DNA oxidation, chromatin compaction, and tiol status seems a good candidate for obtaining a complete picture of pig sperm nucleus status.
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- 2024
21. SUMOylation modulates eIF5A activities in both yeast and pancreatic ductal adenocarcinoma cells
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Conferencia de Rectores de las Universidades Españolas, Consejo Superior de Investigaciones Científicas (España), European Commission, Agencia Estatal de Investigación (España), Ministerio de Ciencia, Innovación y Universidades (España), Ministerio de Ciencia e Innovación (España), Xunta de Galicia, Generalitat Valenciana, Banco Santander, Universidad de Santiago de Compostela, Instituto de Salud Carlos III, Asociación Española Contra el Cáncer, Seoane, Rocío, Lama-Díaz, Tomás, Romero, Antonia María, El Motiam, Ahmed, Martínez-Férriz, Arantxa, Vidal, Santiago, Bouzaher, Yanis H., Blanquer, María, Tolosa, Rocío M., Castillo Mewa, Juan, Álvarez-Rodríguez, Manuel, García-Sastre, Adolfo, Xirodimas, Dimitris, Sutherland, James D., Barrio, Rosa, Alepuz, Paula, Blanco, Miguel G., Farràs, Rosa, Rivas, Carmen, Conferencia de Rectores de las Universidades Españolas, Consejo Superior de Investigaciones Científicas (España), European Commission, Agencia Estatal de Investigación (España), Ministerio de Ciencia, Innovación y Universidades (España), Ministerio de Ciencia e Innovación (España), Xunta de Galicia, Generalitat Valenciana, Banco Santander, Universidad de Santiago de Compostela, Instituto de Salud Carlos III, Asociación Española Contra el Cáncer, Seoane, Rocío, Lama-Díaz, Tomás, Romero, Antonia María, El Motiam, Ahmed, Martínez-Férriz, Arantxa, Vidal, Santiago, Bouzaher, Yanis H., Blanquer, María, Tolosa, Rocío M., Castillo Mewa, Juan, Álvarez-Rodríguez, Manuel, García-Sastre, Adolfo, Xirodimas, Dimitris, Sutherland, James D., Barrio, Rosa, Alepuz, Paula, Blanco, Miguel G., Farràs, Rosa, and Rivas, Carmen
- Abstract
The eukaryotic translation initiation protein eIF5A is a highly conserved and essential factor that plays a critical role in different physiological and pathological processes including stress response and cancer. Different proteomic studies suggest that eIF5A may be a small ubiquitin-like modifier (SUMO) substrate, but whether eIF5A is indeed SUMOylated and how relevant is this modification for eIF5A activities are still unknown.
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- 2024
22. Evaluation of Different Thawing Protocols on Iberian Boar Sperm Preserved for 10 Years at Different Liquid Nitrogen Levels
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European Commission, de Mercado, Eduardo [0000-0003-0321-2431], Álvarez-Rodríguez, Manuel, Tomás-Almenar, Cristina, Nieto-Cristóbal, Helena, de Mercado, Eduardo, European Commission, de Mercado, Eduardo [0000-0003-0321-2431], Álvarez-Rodríguez, Manuel, Tomás-Almenar, Cristina, Nieto-Cristóbal, Helena, and de Mercado, Eduardo
- Abstract
Simple Summary: It remains unclear whether the prolonged preservation of cells, such as sperm, in liquid nitrogen (LN2) leads to their deterioration over time. In this study, we investigated whether the conservation of Iberian pig semen for ten years results in a decline in quality and whether the evaporation of LN2 from the tanks between refills could be a contributing factor. Furthermore, the only way to improve already-frozen samples is to implement specific thawing protocols. Thus, our findings suggest that only samples consistently submerged in LN2 benefit the most from the application of thawing protocols, such as rapid thawing at 70 °C for 8 s. While partial conservation in LN2 and LN2 vapors does not result in a loss of quality over time, it does cause damage that thawing protocols cannot mitigate. This finding suggests that, independently of the lack of the effect of the level of storage in the tank, careful handling and optimization of thawing protocols are essential for the quality preservation of the cryopreserved sperm samples., The conservation of genetic resources in pig breeds, notably the Iberian pig, is crucial for genetic improvement and sustainable production. Prolonged storage in liquid nitrogen (LN2) is recognized for preserving genetic diversity, but potential adverse effects on seminal quality remain debated. This study aims to assess the impact of ten years of storage at different LN2 levels and to optimize thawing protocols for Iberian pig sperm. Sperm samples from 53 boars were cryopreserved and stored at varying LN2 levels and, a decade later, the samples were thawed at 37 °C for 20 s or at 70 °C for 8 s. Sperm motility, membrane integrity, acrosome status, and DNA fragmentation were evaluated in year 0 and year 10. Overall, no significant differences were observed in post-thaw sperm quality between storage levels in year 0 or year 10. But thawing at 70 °C 8 s showed significant improvements, particularly in samples that were always stored in LN2, in all analyzed parameters except fragmentation, which was not affected by cryostorage. This study suggests that the long-term preservation of Iberian pig sperm does not affect quality over time, regardless of whether the samples were fully submerged in LN2. Furthermore, it is determined that thawing at 70 °C for 8 s maximizes post-thaw sperm quality, especially in those samples stored constantly submerged in LN2.
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- 2024
23. Expression of Aquaglyceroporins in Spermatozoa from Wild Ruminants Is Influenced by Photoperiod and Thyroxine Concentrations
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Caballero, Beatriz, Santiago-Moreno, Julián, Pequeño, Belén, Martínez Madrid, Carmen Belén, Castaño, Cristina, Bóveda, Paula, Velázquez, Rosario, Toledano-Díaz, Adolfo, Álvarez-Rodríguez, Manuel, Rodríguez-Martínez, Heriberto, Caballero, Beatriz, Santiago-Moreno, Julián, Pequeño, Belén, Martínez Madrid, Carmen Belén, Castaño, Cristina, Bóveda, Paula, Velázquez, Rosario, Toledano-Díaz, Adolfo, Álvarez-Rodríguez, Manuel, and Rodríguez-Martínez, Heriberto
- Abstract
J. Santiago-Moreno recibió una beca del Ministerio de Ciencia, Innovación y Universidades para visita de investigadores seniors a centros extranjeros (PRX19/00149). El Consejo Sueco de Investigación FORMAS (2017-00946 y 2019-00288), Estocolmo, Suecia, proporcionó apoyo analítico., This work identified the presence of AQPs in frozen-thawed sperm of wild ruminants and assessed the influence of the interaction between photoperiod and thyroxine on AQP expression, and on testosterone secretion. Thyroxine and melatonin were administered to ibexes. In a second experiment, performed in mouflons, circulating thyroxine was reduced via treatment with propylthiouracil (PTU), and an artificial long day (LD) photoperiod established. In the ibexes, the melatonin treatment increased the blood plasma testosterone concentration, reduced the cryoresistance ratio (CR) for sperm viability and the presence of an intact acrosome, and increased the percentage of sperm with AQP7 in the acrosome and of AQP3 and AQP10 in the midpiece. In the mouflons, neither the PTU treatment, the LD, nor the combination of both affected the CR of any sperm variable. The percentage of sperm with AQP3 increased in the post-acrosome region but decreased in the tail in the LD+PTU group. The percentage of sperm with AQP10 in the principal piece and endpiece was lower in the PTU+LD group than in the control and LD groups. The influence of photoperiod/melatonin on AQP expression might be indirectly exerted through changes in the testosterone concentration, and thus ultimately affect sperm cryoresistance., Agencia Estatal de Investigación (AEI) (AGL2017-85753-R y PID2020-113288RB-I00/AEI/10.13039/501100011033), MCINN (AGL2017-85753-R y PID2020-113288RB-I00/AEI/10.13039/501100011033), FEDER/EU, Depto. de Medicina y Cirugía Animal, Fac. de Veterinaria, TRUE, pub
- Published
- 2024
24. Sperm freezability is neither associated with the expression of aquaporin 3 nor sperm head dimensions in dromedary camel (Camelus dromedarius)
- Author
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O'Brien, Emma, Malo, Clara, Castaño, Cristina, García-Casado, Pedro, Toledano-Díaz, Adolfo, Martínez Madrid, Carmen Belén, Rodriguez-Martinez, Heriberto, Álvarez-Rodríguez, Manuel, Santiago-Moreno, Julián, O'Brien, Emma, Malo, Clara, Castaño, Cristina, García-Casado, Pedro, Toledano-Díaz, Adolfo, Martínez Madrid, Carmen Belén, Rodriguez-Martinez, Heriberto, Álvarez-Rodríguez, Manuel, and Santiago-Moreno, Julián
- Abstract
Este trabajo también fue financiado por un contrato entre Zoitechlab S.L. (Arquimea Group) y el INIA (CON18-141), The expression of aquaglyceroporin 3 (AQP-3) has been demonstrated in the spermatozoa of several mammalian species and its role has been associated with cryotolerance. Post-thaw sperm quality from individual dromedary males with different response to freezing-thawing process was evaluated through sperm head morphometry. In order to understand the cellular mechanisms affected by cryoinjury we have explored the presence and distribution of sperm AQP-3 using western blotting and immunocytochemistry. WB showed different intensity of the specific signal bands at 28 kDa. Immunofluorescence assessments allowed us to identify five different and clear AQP-3 distribution patterns of labelling in the sperm plasma membrane; acrosome, post-acrosome, mid-piece, and principal and final tail. Although expression of AQP-3 varied among male ejaculates, the individual sperm response to freeze-thawing was not associated with AQP-3 expression. Thus, AQP3 expressions do not seem like a reliable predictor of sperm response to freeze-thawing process in this species. This work is the first to describe the morphometric characteristics of the heads of dromedary spermatozoa. No correlation was found between sperm head dimensions and sperm quality variables after freeze-thawing suggesting that dromedary camel sperm head morphometry is also not a reliable predictor of cryosurvival., Agencia Estatal de Investigación (AEI) (AGL2017-85753-R y PID2020-113288RB-100/AEI/10.13039/501100011033), MCINN/AEI/FEDER (AGL2017-85753-R y PID2020-113288RB-100/AEI/10.13039/501100011033), EU (AGL2017-85753-R y PID2020-113288RB-100/AEI/10.13039/501100011033), Depto. de Medicina y Cirugía Animal, Fac. de Veterinaria, TRUE, pub
- Published
- 2024
25. Effect of quercetin loaded liposomes or nanostructured lipid carrier (NLC) on post-thawed sperm quality and fertility of rooster sperm
- Author
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Najafi, Abouzar, Kia, Hossein Daghigh, Mehdipour, Mahdieh, Hamishehkar, Hamed, and Álvarez-Rodríguez, Manuel
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- 2020
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- View/download PDF
26. In vitro assessment of egg yolk-, soya bean lecithin- and liposome-based extenders for cryopreservation of dairy bull semen
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Miguel-Jimenez, Sara, Rivera del Alamo, Maria Montserrat, Álvarez-Rodríguez, Manuel, Hidalgo, Carlos Olegario, Peña, Ana Isabel, Muiño, Rodrigo, Rodríguez-Gil, Joan Enric, and Mogas, Teresa.
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- 2020
- Full Text
- View/download PDF
27. Inclusion of novel additives on extenders to improve rabbit sperm cryopreservation
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Gardela, Jaume, primary, Ruiz-Conca, Mateo, additional, Palomares, Anna, additional, Olvera-Maneu, Sergi, additional, García-Calvo, Laura, additional, López-Béjar, Manel, additional, Martinez-Pastor, Felipe, additional, and Álvarez-Rodríguez, Manuel, additional
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- 2023
- Full Text
- View/download PDF
28. Bicarbonate and BSA increase the capacitation pattern and acrosomal exocytosis in boar sperm after 120 min of incubation
- Author
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Dudkiewicz, Sylwia, primary, Peris‐Frau, Patricia, additional, Nieto‐Cristóbal, Helena, additional, Santiago‐Moreno, Julián, additional, de Mercado, Eduardo, additional, and Álvarez‐Rodríguez, Manuel, additional
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- 2023
- Full Text
- View/download PDF
29. Exosomes in specific fractions of the boar ejaculate contain CD44: A marker for epididymosomes?
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Alvarez-Rodriguez, Manuel, Ljunggren, Stefan A., Karlsson, Helen, and Rodriguez-Martinez, Heriberto
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- 2019
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30. Mating modifies the expression of crucial oxidative-reductive transcripts in the pig oviductal sperm reservoir: is the female ensuring sperm survival?
- Author
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Ministerio de Ciencia e Innovación (España), Agencia Estatal de Investigación (España), Ministerio de Economía y Competitividad (España), European Commission, Álvarez-Rodríguez, Manuel [0000-0003-0120-354X], Martinez, Emilio A. [0000-0003-1260-9721], Rodriguez-Martinez, Heriberto [0000-0002-5194-2124], Álvarez-Rodríguez, Manuel, Roca, Jordi, Martinez, Emilio A., Rodriguez-Martinez, Heriberto, Ministerio de Ciencia e Innovación (España), Agencia Estatal de Investigación (España), Ministerio de Economía y Competitividad (España), European Commission, Álvarez-Rodríguez, Manuel [0000-0003-0120-354X], Martinez, Emilio A. [0000-0003-1260-9721], Rodriguez-Martinez, Heriberto [0000-0002-5194-2124], Álvarez-Rodríguez, Manuel, Roca, Jordi, Martinez, Emilio A., and Rodriguez-Martinez, Heriberto
- Abstract
Mating induces large changes in the female genital tract, warranting female homeostasis and immune preparation for pregnancy, including the preservation of crucial oxidative status among its pathways. Being highly susceptible to oxidative stress, sperm survival and preserved function depend on the seminal plasma, a protection that is removed during sperm handling but also after mating when spermatozoa enter the oviduct. Therefore, it is pertinent to consider that the female sperm reservoir takes up this protection, providing a suitable environment for sperm viability. These aspects have not been explored despite the increasing strategies in modulating the female status through diet control and nutritional supplementation.
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- 2023
31. Variation of existence and location of aquaporin 3 in relation to cryoresistance of ram spermatozoa
- Author
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Agencia Estatal de Investigación (España), Pequeño, Belén [0000-0003-3962-1982], Castaño, Cristina [0000-0003-1134-1436], Álvarez-Rodríguez, Manuel [0000-0003-0120-354X], Bóveda, Paula [0000-0001-5749-4345], Millán de la Blanca, María Gemma [0000-0002-2810-8510], Toledano Díaz, Adolfo [0000-0001-6679-485X], Galarza, Diego Andres [0000-0002-0266-5431], Rodriguez-Martinez, Heriberto [0000-0002-5194-2124], Martínez-Madrid, Belén [0000-0001-6852-4597], Santiago Moreno, Julián [0000-0001-5551-8120], Pequeño, Belén, Castaño, Cristina, Álvarez-Rodríguez, Manuel, Bóveda, Paula, Millán de la Blanca, María Gemma, Toledano-Díaz, A., Galarza, Diego Andres, Rodriguez-Martinez, Heriberto, Martínez-Madrid, Belén, Santiago Moreno, Julián, Agencia Estatal de Investigación (España), Pequeño, Belén [0000-0003-3962-1982], Castaño, Cristina [0000-0003-1134-1436], Álvarez-Rodríguez, Manuel [0000-0003-0120-354X], Bóveda, Paula [0000-0001-5749-4345], Millán de la Blanca, María Gemma [0000-0002-2810-8510], Toledano Díaz, Adolfo [0000-0001-6679-485X], Galarza, Diego Andres [0000-0002-0266-5431], Rodriguez-Martinez, Heriberto [0000-0002-5194-2124], Martínez-Madrid, Belén [0000-0001-6852-4597], Santiago Moreno, Julián [0000-0001-5551-8120], Pequeño, Belén, Castaño, Cristina, Álvarez-Rodríguez, Manuel, Bóveda, Paula, Millán de la Blanca, María Gemma, Toledano-Díaz, A., Galarza, Diego Andres, Rodriguez-Martinez, Heriberto, Martínez-Madrid, Belén, and Santiago Moreno, Julián
- Abstract
Osmotic changes during the process of freeze-thawing involve changes in the location of aquaporins (AQPs) in membrane domains of spermatozoa. Some AQPs, like aquaporin 3 (AQP3), are linked to sperm cryotolerance in the porcine species. Conspicuous individual variability exists between rams and their ejaculates, which may be classified as displaying good freezability (GFE) or poor freezability (PFE), depending on several endogenous and environmental factors. The present work aimed to examine whether differences in freezability could even involve changes in location and expression of AQP3 in ram spermatozoa.
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- 2023
32. Editorial: Cryopreservation of mammalian gametes and embryos: implications of oxidative and nitrosative stress and potential role of antioxidants
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National Key Research and Development Program (China), Ofosu, Jones [0000-0003-4884-1629], Sun, Xiuzhu [0000-0002-9760-0584], Álvarez-Rodríguez, Manuel [0000-0003-0120-354X], Zhou, Guangbin [0000-0002-4493-8311], Ofosu, Jones, Zhang, Yunhai, Liu, Ying, Sun, Xiuzhu, Quan, Guobo, Álvarez-Rodríguez, Manuel, Zhou, Guangbin, National Key Research and Development Program (China), Ofosu, Jones [0000-0003-4884-1629], Sun, Xiuzhu [0000-0002-9760-0584], Álvarez-Rodríguez, Manuel [0000-0003-0120-354X], Zhou, Guangbin [0000-0002-4493-8311], Ofosu, Jones, Zhang, Yunhai, Liu, Ying, Sun, Xiuzhu, Quan, Guobo, Álvarez-Rodríguez, Manuel, and Zhou, Guangbin
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- 2023
33. Identification and quantification of tyrosine phosphorylated proteins induced by FERT medium during rabbit sperm capacitation at different times
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Agencia Estatal de Investigación (España), Ministerio de Economía y Competitividad (España), European Commission, Gimeno-Martos, Silvia [0000-0002-0455-3409], Arias-Álvarez, María [0000-0001-7907-7454], Lorenzo, Pedro L. [0000-0001-7955-4206], Álvarez-Rodríguez, Manuel [0000-0003-0120-354X], García-García, Rosa M. [0000-0002-3393-8906], Gimeno-Martos, Silvia, Arias-Álvarez, María, Quiroga, Alejandra C., Lorenzo, Pedro L., Álvarez-Rodríguez, Manuel, Rebollar, P. G., García-García, Rosa M., Agencia Estatal de Investigación (España), Ministerio de Economía y Competitividad (España), European Commission, Gimeno-Martos, Silvia [0000-0002-0455-3409], Arias-Álvarez, María [0000-0001-7907-7454], Lorenzo, Pedro L. [0000-0001-7955-4206], Álvarez-Rodríguez, Manuel [0000-0003-0120-354X], García-García, Rosa M. [0000-0002-3393-8906], Gimeno-Martos, Silvia, Arias-Álvarez, María, Quiroga, Alejandra C., Lorenzo, Pedro L., Álvarez-Rodríguez, Manuel, Rebollar, P. G., and García-García, Rosa M.
- Abstract
Rabbit sperm capacitation is still a largely unknown process. Thus, the main objetive of this study was to examine the molecular mechanisms that modulate capacitation in this species at different times.
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- 2023
34. Location of aquaporins 3, 7 and 10 in frozen-thawed ejaculated and cauda epididymal spermatozoa from the Iberian ibex, mouflon, and chamois
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Agencia Estatal de Investigación (España), European Commission, Ministerio de Ciencia e Innovación (España), Pequeño, Belén [0000-0003-3962-1982], Martínez-Madrid, Belén [0000-0001-6852-4597], Castaño, Cristina [0000-0003-1134-1436], Toledano-Díaz, A. [0000-0001-6679-485X], Bóveda, Paula [0000-0001-5749-4345], Esteso, Milagros C. [0000-0002-8963-5736], Gómez-Guillamón, Félix [0000-0001-9757-9589], Álvarez-Rodríguez, Manuel [0000-0003-0120-354X], Rodriguez-Martinez, Heriberto [0000-0002-5194-2124], Santiago Moreno, Julián [0000-0001-5551-8120], Pequeño, Belén, Martínez-Madrid, Belén, Castaño, Cristina, Toledano-Díaz, A., Bóveda, Paula, Esteso, Milagros C., Gómez-Guillamón, Félix, Prieto, Paloma, Marcos-Beltrán, J. L., Álvarez-Rodríguez, Manuel, Rodriguez-Martinez, Heriberto, Santiago Moreno, Julián, Agencia Estatal de Investigación (España), European Commission, Ministerio de Ciencia e Innovación (España), Pequeño, Belén [0000-0003-3962-1982], Martínez-Madrid, Belén [0000-0001-6852-4597], Castaño, Cristina [0000-0003-1134-1436], Toledano-Díaz, A. [0000-0001-6679-485X], Bóveda, Paula [0000-0001-5749-4345], Esteso, Milagros C. [0000-0002-8963-5736], Gómez-Guillamón, Félix [0000-0001-9757-9589], Álvarez-Rodríguez, Manuel [0000-0003-0120-354X], Rodriguez-Martinez, Heriberto [0000-0002-5194-2124], Santiago Moreno, Julián [0000-0001-5551-8120], Pequeño, Belén, Martínez-Madrid, Belén, Castaño, Cristina, Toledano-Díaz, A., Bóveda, Paula, Esteso, Milagros C., Gómez-Guillamón, Félix, Prieto, Paloma, Marcos-Beltrán, J. L., Álvarez-Rodríguez, Manuel, Rodriguez-Martinez, Heriberto, and Santiago Moreno, Julián
- Abstract
Spermatozoa collected from the cauda epididymis of wild ruminants are more cryoresistant than ejaculated spermatozoa. Changes in the membrane location of aquaporins (AQPs) follow the osmotic changes that occur during freeze-thawing, and might influence the cryosurvival of spermatozoa depending on their source. This work reports the location of AQP3, AQP7 and AQP10 in the cauda epididymal and post-ejaculation spermatozoa of three wild mountain ungulate species (Iberian ibex, mouflon, and chamois), as determined by Western blotting (WB) and immunocytochemistry (ICC) using commercial rabbit polyclonal primary antibodies. WB confirmed the presence of all three AQPs in the spermatozoa of all the studied species, while ICC showed AQP3 to be mainly located in the sperm acrosome, mid-piece, principal piece, and end piece, both in cauda epididymal and ejaculated cells. The percentage of ejaculated spermatozoa showing AQP3 in the principal piece was higher in the ibex than in the chamois (P < 0.05), and higher in epididymal spermatozoa in the mouflon than in the chamois (P < 0.05). AQP7 was located in the acrosome of both epididymal and ejaculated spermatozoa, as well as in the cytoplasmic droplet of the epididymal spermatozoa of all three species. No differences were seen between the species with respect to the percentage of spermatozoa showing AQP7. AQP10 was located mainly in the mid-piece, principal piece and end piece of the sperm tail in both epididymal and ejaculated spermatozoa. The percentage of mouflon spermatozoa with AQP10 in the end piece was higher in the cauda epididymal than in the ejaculated spermatozoa (P < 0.05). In conclusion, except for AQP10 in the mouflon, the locations of the studied AQPs are similar in epididymal and ejaculated spermatozoa, with inter-species differences seen only for AQP3. Further studies are needed to determine what this might mean with respect to sperm cryopreservation.
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- 2023
35. Evaluation of the progesterone inhibitor RU486 in boar sperm cryopreservation
- Author
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Agencia Estatal de Investigación (España), Ministerio de Ciencia e Innovación (España), European Commission, de Mercado, Eduardo [0000-0003-0321-2431], Nieto-Cristóbal, Helena [0000-0003-1192-9467], Álvarez-Rodríguez, Manuel [0000-0003-0120-354X], Gala, Nerea, de Mercado, Eduardo, Nieto-Cristóbal, Helena, Álvarez-Rodríguez, Manuel, Agencia Estatal de Investigación (España), Ministerio de Ciencia e Innovación (España), European Commission, de Mercado, Eduardo [0000-0003-0321-2431], Nieto-Cristóbal, Helena [0000-0003-1192-9467], Álvarez-Rodríguez, Manuel [0000-0003-0120-354X], Gala, Nerea, de Mercado, Eduardo, Nieto-Cristóbal, Helena, and Álvarez-Rodríguez, Manuel
- Abstract
Boar sperm cryopreservation is a sub-optimal method due to the high sensitivity of sperm to cold shock. In addition, sperm cryopreservation induces capacitation-like changes, such as sperm membrane modifications, thereby affecting calcium channels, which causes variations in the ion exchange. These structural changes and intracellular signalling lead to increased acrosomal reactions or different patterns of movement, called cryo-capacitation. On the other hand, progesterone (P4) is an inductor of capacitation, including acrosomal reaction, mainly by inducing changes in the plasmatic membrane that allow the entry of calcium, which in turn activates the signalling pathway of adenylate cyclase and the production of cAMP. Being said that the use of a specific progesterone inhibitor could revert the P4 action, this preventing capacitationlike changes that happen during sperm cryopreservation. The aim of this study was to evaluate the effect of different concentrations of a progesterone inhibitor (RU486) in the freezing extender, on post-thaw boar sperm quality. Throughout the experiment, commercial artificial insemination semen doses from six fertile boars were centrifuged and diluted (300 x 106 sperm/mL) in a freezing extender (LEY: 20% egg yolk and 80% lactose), supplemented with RU486 (A: non-supplemented (control); B: 5 mM; C: 10 mM; D: 20 mM). The samples were cooled to 5 °C, extended (1:3; v:v) in LEYGO (LEY + 9% glycerol and 1.5% Equex) and 0.5 mL straws frozen in LN2 vapours, and thawed at 37 °C/20 sec. Total motile sperm (%TMS) and other kinetics parameters were evaluated with the IA Station CASA system (SPERMTECH®). Furthermore, membrane integrity and acrosome damage were determined by using triple staining (Hoechst 33342/propidium iodide PI/PNA-FITC) under epifluorescence microscopy. Sperm with undamaged membrane and acrosome (%VIAB) or reacted acrosome (%VIAB/AR) and sperm with damaged membrane and acrosome (%DEAD/AR) were recorded. Moreover, sperm with high
- Published
- 2023
36. Pre-freezing selection of Holstein bull semen with the BoviPure colloid as double- or single-layer centrifugation improves the post-thawing quality
- Author
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Ministerio de Economía y Competitividad (España), Ministerio de Ciencia e Innovación (España), Agencia Estatal de Investigación (España), European Commission, Salman, A. [0000-0003-4960-9020], Fernández-Alegre, Estela [0000-0002-2223-1788], Domínguez, Juan C. [0000-0003-3654-1573], Álvarez-Rodríguez, Manuel [0000-0003-0120-354X], Caamaño, J. N. [0000-0001-6449-5784], Martínez-Pastor, F. [0000-0003-2987-4302], Fernández-Fernández, Alejandro [0000-0003-2524-3597], Areán-Dablanca, Héctor [0000-0002-6811-8983], Salman, A., Fernández-Alegre, Estela, Francisco-Vázquez, Rubén, Domínguez, Juan C., Álvarez-Rodríguez, Manuel, Caamaño, J. N., Martínez-Pastor, F., Gómez-Martín, Rubén, Fernández-Fernández, Alejandro, Areán-Dablanca, Héctor, Ministerio de Economía y Competitividad (España), Ministerio de Ciencia e Innovación (España), Agencia Estatal de Investigación (España), European Commission, Salman, A. [0000-0003-4960-9020], Fernández-Alegre, Estela [0000-0002-2223-1788], Domínguez, Juan C. [0000-0003-3654-1573], Álvarez-Rodríguez, Manuel [0000-0003-0120-354X], Caamaño, J. N. [0000-0001-6449-5784], Martínez-Pastor, F. [0000-0003-2987-4302], Fernández-Fernández, Alejandro [0000-0003-2524-3597], Areán-Dablanca, Héctor [0000-0002-6811-8983], Salman, A., Fernández-Alegre, Estela, Francisco-Vázquez, Rubén, Domínguez, Juan C., Álvarez-Rodríguez, Manuel, Caamaño, J. N., Martínez-Pastor, F., Gómez-Martín, Rubén, Fernández-Fernández, Alejandro, and Areán-Dablanca, Héctor
- Abstract
Artificial insemination (AI) is critical for breeding in the dairy industry. High-merit bulls can present low freezability, hampering genetic dissemination. Thawed semen can be improved using density gradient centrifugation (DGC) with colloids, but little information deals with the pre-freezing application. Thus, the BoviPure colloid (optimized for bull spermatozoa) was tested for pre-freezing application as the usual double-layer (DLC) versus single-layer (SLC, quick and economical). Semen from twelve Holstein-Friesian bulls was extended with OPTIXcell extender, frozen (Control), or processed by SLC or DLC and frozen. Sperm were assessed pre-freezing for motility and viability and post-thawing (directly and after 4 h 38 °C) for apoptosis, capacitation status, acrosomal damage, mitochondrial activity, cytoplasmic and mitochondrial reactive oxygen species (ROS), and chromatin status (SCSA for DNA fragmentation and chromatin compaction and monobromobimane, mBBr, for disulfide bridges evaluation). The DGC improved parameters post-thawing (e.g., 57.5%±10.1 motility vs. control 53.3% ± 11.2) at the cost of sperm loss (sperm recovery of DGC 14.4% ± 2.5 and SLC 17.4% ± 2.5). DNA fragmentation (%DFI) decreased (0.21% ± 0.53 vs. control 1.30% ± 0.10), and SLC reduced chromatin compaction. A clustering procedure separated lesser (LF) and greater freezability (GF) bulls. LF samples were especially benefited by DGC, with SLC providing better post-thawing results for this group. In conclusion, pre-freezing DGC improved sperm parameters post-thawing, potentially improving the cryopreservation of low-freezability semen from high-merit bulls. SLC, quicker and economical, would be preferable since it showed similar or higher performance than DLC.
- Published
- 2023
37. Immunolocalisation of aquaporins 3, 7, 9 and 10 in the epididymis of three wild ruminant species (Iberian ibex, mouflon and chamois) and sperm cryoresistance
- Author
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Agencia Estatal de Investigación (España), Martínez-Madrid, Belén [0000-0001-6852-4597], Martínez-Cáceres, Carlos [0000-0003-3307-1326], Pequeño, Belén [0000-0003-3962-1982], Castaño, Cristina [0000-0003-1134-1436], Toledano-Diaz, A. [0000-0001-6679-485X], Bóveda, Paula [0000-0001-5749-4345], Álvarez-Rodríguez, Manuel [0000-0003-0120-354X], Rodriguez-Martinez, Heriberto [0000-0002-5194-2124], Santiago Moreno, Julián [0000-0001-5551-8120], Martínez-Madrid, Belén, Martínez-Cáceres, Carlos, Pequeño, Belén, Castaño, Cristina, Toledano-Díaz, A., Bóveda, Paula, Prieto, Paloma, Álvarez-Rodríguez, Manuel, Rodriguez-Martinez, Heriberto, Santiago Moreno, Julián, Agencia Estatal de Investigación (España), Martínez-Madrid, Belén [0000-0001-6852-4597], Martínez-Cáceres, Carlos [0000-0003-3307-1326], Pequeño, Belén [0000-0003-3962-1982], Castaño, Cristina [0000-0003-1134-1436], Toledano-Diaz, A. [0000-0001-6679-485X], Bóveda, Paula [0000-0001-5749-4345], Álvarez-Rodríguez, Manuel [0000-0003-0120-354X], Rodriguez-Martinez, Heriberto [0000-0002-5194-2124], Santiago Moreno, Julián [0000-0001-5551-8120], Martínez-Madrid, Belén, Martínez-Cáceres, Carlos, Pequeño, Belén, Castaño, Cristina, Toledano-Díaz, A., Bóveda, Paula, Prieto, Paloma, Álvarez-Rodríguez, Manuel, Rodriguez-Martinez, Heriberto, and Santiago Moreno, Julián
- Abstract
In the epididymis, epithelial cells manage changes in the luminal environment for proper sperm maturation. Moreover, aquaglyceroporins, a subgroup of aquaporins (AQP), modulate the transport of water, glycerol and other small molecules in epithelial cells.
- Published
- 2023
38. The Cation/Calcium Channel of Sperm (CatSper): A Common Role Played Despite Inter-Species Variation?
- Author
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Vicente-Carrillo, Alejandro, primary, Álvarez-Rodríguez, Manuel, additional, and Rodriguez-Martinez, Heriberto, additional
- Published
- 2023
- Full Text
- View/download PDF
39. Editorial: Cryopreservation of mammalian gametes and embryos: implications of oxidative and nitrosative stress and potential role of antioxidants
- Author
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Ofosu, Jones, Zhang, Yunhai, Liu, Ying, Sun, Xiuzhu, Quan, Guobo, Álvarez-Rodríguez, Manuel, Zhou, Guangbin, National Key Research and Development Program (China), Ofosu, Jones, Sun, Xiuzhu, Álvarez-Rodríguez, Manuel, and Zhou, Guangbin
- Subjects
Cryopreservation ,Oocyte ,General Veterinary ,Embryo ,Reactive oxygen species ,Antioxidants ,Sperm - Abstract
4 Pág., This work was financially supported by the National Key Research and Development Program of China (Grant No. 2021YFD1200403).
- Published
- 2023
40. Effect of Honey, Coenzyme Q10, and β-Carotene/α-Tocopherol as Novel Additives in Rabbit-Sperm Cryopreservation Extender
- Author
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Gardela, Jaume, primary, Ruiz-Conca, Mateo, additional, Palomares, Anna, additional, Olvera-Maneu, Sergi, additional, García-Calvo, Laura, additional, López-Béjar, Manel, additional, Martínez-Pastor, Felipe, additional, and Álvarez-Rodríguez, Manuel, additional
- Published
- 2023
- Full Text
- View/download PDF
41. Mating modifies the expression of crucial oxidative-reductive transcripts in the pig oviductal sperm reservoir: is the female ensuring sperm survival?
- Author
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Álvarez-Rodríguez, Manuel, primary, Roca, Jordi, additional, Martínez, Emilio A., additional, and Rodríguez-Martínez, Heriberto, additional
- Published
- 2023
- Full Text
- View/download PDF
42. The CatSper channel modulates boar sperm motility during capacitation
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Vicente-Carrillo, Alejandro, Álvarez-Rodríguez, Manuel, and Rodríguez-Martínez, Heriberto
- Published
- 2017
- Full Text
- View/download PDF
43. Ethylene glycol, but not DMSO, could replace glycerol inclusion in soybean lecithin-based extenders in ram sperm cryopreservation
- Author
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Najafi, Abouzar, Daghigh-Kia, Hossein, Dodaran, Hossein Vaseghi, Mehdipour, Mahdieh, and Alvarez-Rodriguez, Manuel
- Published
- 2017
- Full Text
- View/download PDF
44. Bicarbonate and BSA increase the capacitation pattern and acrosomal exocytosis in boar sperm after 120 min of incubation.
- Author
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Dudkiewicz, Sylwia, Peris‐Frau, Patricia, Nieto‐Cristóbal, Helena, Santiago‐Moreno, Julián, de Mercado, Eduardo, and Álvarez‐Rodríguez, Manuel
- Subjects
BICARBONATE ions ,EXOCYTOSIS ,SPERMATOZOA ,BOARS ,MITOCHONDRIAL membranes ,SEMEN ,MEMBRANE potential - Abstract
Sperm capacitation is a crucial step towards the acquisition of fertilizing capacity. Despite the attempts to mimic the in vivo situation, there is still a lack of standardization in vitro techniques. Bicarbonate and serum albumin (BSA) are routinely used, although controversial results are reported regarding the optimal concentration of each compound. In addition, whether caffeine is needed on in vitro capacitation media in boar sperm remains to be elucidated. Here, 18 boar commercial artificial insemination doses were used to test different concentrations of bicarbonate (19, 37 or 56 mM) in experiment 1, BSA (1.5, 3, 4.5 mg/mL) in experiment 2 and the presence or absence of caffeine (5.15 mM) experiment 3. We analysed at 0, 30 and 120 min of incubation at 38.5°C, 5% CO2: Total motility (TMOT), membrane integrity (VIAB), acrosomal exocytosis (rAcro; H33342/PI/PNA), capacitation status (chlortetracycline staining CTC) and mitochondrial membrane potential (JC‐1). The higher concentrations of bicarbonate (37 and 56 mM) decreased TM and VIAB (p <.01) but increased rAcro (p <.01) after 120 min of incubation compared to the fresh control. In contrast, only the BSA concentration of 3 mg/mL reduced the VIAB at 120 min, but all the concentrations tested increased the average of JC‐1 and decreased TM (p <.01) throughout incubation compared to the fresh control. Finally, in experiment 3, when boar sperm were incubated in the capacitating media with bicarbonate, BSA and with or without caffeine, the capacitated pattern measured by the CTC technique and rAcro increased after 120 min of incubation (p <.01) compared to fresh control, either in the presence or in the absence of caffeine. In summary, our results suggested that the combination of capacitating components, like bicarbonate and BSA, contributed to increasing the proportion of capacitated boar spermatozoa, mitochondrial membrane potential as well as acrosomal exocytosis. However, caffeine did not significantly influence in vitro sperm capacitation in this species. [ABSTRACT FROM AUTHOR]
- Published
- 2024
- Full Text
- View/download PDF
45. Editorial: New advances in our understanding of fertilization
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Vicente-Carrillo, Alejandro [0000-0002-0256-1958], Casao,Adriana [0000-0003-1997-4262], Álvarez-Rodríguez, Manuel [0000-0003-0120-354X], Vicente-Carrillo, Alejandro, Casao, Adriana, Álvarez-Rodríguez, Manuel, Vicente-Carrillo, Alejandro [0000-0002-0256-1958], Casao,Adriana [0000-0003-1997-4262], Álvarez-Rodríguez, Manuel [0000-0003-0120-354X], Vicente-Carrillo, Alejandro, Casao, Adriana, and Álvarez-Rodríguez, Manuel
- Published
- 2022
46. Sperm freezability is neither associated with the expression of aquaporin 3 nor sperm head dimensions in dromedary camel (Camelus dromedarius)
- Author
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Ministerio de Ciencia e Innovación (España), Agencia Estatal de Investigación (España), European Commission, Malo, Clara [0000-0003-2835-4823], Castaño, Cristina [0000-0003-1134-1436], Toledano Díaz, Adolfo [0000-0001-6679-485X], Martínez-Madrid, Belén [0000-0001-6852-4597], Rodriguez-Martinez, Heriberto [0000-0002-5194-2124], Álvarez-Rodríguez, Manuel [0000-0003-0120-354X], Santiago Moreno, Julián [0000-0001-5551-8120], O'Brien, Emma, Malo, Clara, Castaño, Cristina, García-Casado, Pedro, Toledano-Díaz, A., Martínez-Madrid, Belén, Rodriguez-Martinez, Heriberto, Álvarez-Rodríguez, Manuel, Santiago Moreno, Julián, Ministerio de Ciencia e Innovación (España), Agencia Estatal de Investigación (España), European Commission, Malo, Clara [0000-0003-2835-4823], Castaño, Cristina [0000-0003-1134-1436], Toledano Díaz, Adolfo [0000-0001-6679-485X], Martínez-Madrid, Belén [0000-0001-6852-4597], Rodriguez-Martinez, Heriberto [0000-0002-5194-2124], Álvarez-Rodríguez, Manuel [0000-0003-0120-354X], Santiago Moreno, Julián [0000-0001-5551-8120], O'Brien, Emma, Malo, Clara, Castaño, Cristina, García-Casado, Pedro, Toledano-Díaz, A., Martínez-Madrid, Belén, Rodriguez-Martinez, Heriberto, Álvarez-Rodríguez, Manuel, and Santiago Moreno, Julián
- Abstract
The expression of aquaglyceroporin 3 (AQP-3) has been demonstrated in the spermatozoa of several mammalian species and its role has been associated with cryotolerance. Post-thaw sperm quality from individual dromedary males with different response to freezing-thawing process was evaluated through sperm head morphometry. In order to understand the cellular mechanisms affected by cryoinjury we have explored the presence and distribution of sperm AQP-3 using western blotting and immunocytochemistry. WB showed different intensity of the specific signal bands at 28 kDa. Immunofluorescence assessments allowed us to identify five different and clear AQP-3 distribution patterns of labelling in the sperm plasma membrane; acrosome, post-acrosome, mid-piece, and principal and final tail. Although expression of AQP-3 varied among male ejaculates, the individual sperm response to freeze-thawing was not associated with AQP-3 expression. Thus, AQP3 expressions do not seem like a reliable predictor of sperm response to freeze-thawing process in this species. This work is the first to describe the morphometric characteristics of the heads of dromedary spermatozoa. No correlation was found between sperm head dimensions and sperm quality variables after freeze-thawing suggesting that dromedary camel sperm head morphometry is also not a reliable predictor of cryosurvival.
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- 2022
47. Changes in aquaporins mRNA expression and liquid storage at 17°C: A potential biomarker of boar sperm quality?
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Ministerio de Ciencia e Innovación (España), Agencia Estatal de Investigación (España), Gardela, Jaume [0000-0001-7524-2088], Ruiz-Conca, Mateo [0000-0001-7092-7993], López-Béjar, Manel [0000-0001-9490-6126], Martinez, Cristina A. [0000-0001-6811-0191], Rodriguez-Martinez, Heriberto [0000-0002-5194-2124], Álvarez-Rodríguez, Manuel [0000-0003-0120-354X], Castany Quintana, Marina, Gardela, Jaume, Ruiz-Conca, Mateo, López-Béjar, Manel, Martinez, Cristina A., Rodriguez-Martinez, Heriberto, Álvarez-Rodríguez, Manuel, Ministerio de Ciencia e Innovación (España), Agencia Estatal de Investigación (España), Gardela, Jaume [0000-0001-7524-2088], Ruiz-Conca, Mateo [0000-0001-7092-7993], López-Béjar, Manel [0000-0001-9490-6126], Martinez, Cristina A. [0000-0001-6811-0191], Rodriguez-Martinez, Heriberto [0000-0002-5194-2124], Álvarez-Rodríguez, Manuel [0000-0003-0120-354X], Castany Quintana, Marina, Gardela, Jaume, Ruiz-Conca, Mateo, López-Béjar, Manel, Martinez, Cristina A., Rodriguez-Martinez, Heriberto, and Álvarez-Rodríguez, Manuel
- Abstract
Artificial insemination (AI) for pigs relies on liquid storage of extended semen at 17°C, which preserves sperm quality and ensures its fertilizing capacity. Routine quality controls include the evaluation of sperm motility, viability and capacitation status. The physiological functions of all these features depend on transmembrane aquaporins (AQPs), proteins playing key roles in osmoadaptation. In this study, we made a relative quantification, using RT-qPCR, of the mRNA of several sperm AQPs in AI-liquid semen doses before and after a 48-hr incubation period, aiming to determine possible quantitative compromising expression changes during the process that could serve as a diagnostic tool. Our results showed a decrease in classical sperm motility variables (total and progressive motility and velocity) and sperm viability after 48-hr storage, whereas capacitation status increased overtime. mRNA expression increased in the orthodox AQP4 and AQP6 after 48-hr incubation, relative to control (0 hr) and 24-hr time-points. Moreover, mRNA expression of aquaglyceroporins AQP3, AQP7 and AQP10 was higher after 48-hr incubation, confirmed by AQP7-protein validation using Western blot. Our results indicate that expression levels of AQPs-mRNA can change in ejaculated pig spermatozoa under conditions of ex-vivo incubation that could modify sperm homeostasis, suggesting it could eventually become a relevant molecular biomarker to assess the efficiency of liquid storage of pig semen.
- Published
- 2022
48. Effect of Honey, Coenzyme Q10, and β-Carotene/α-Tocopherol as Novel Additives in Rabbit-Sperm Cryopreservation Extender
- Author
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Gardela, Jaume, Ruiz-Conca, Mateo, Palomares, Anna, Olvera-Maneu, Sergi, García-Calvo, Laura, López-Béjar, Manel, Martínez-Pastor, Felipe, Álvarez-Rodríguez, Manuel, Gardela, Jaume, Ruiz-Conca, Mateo, Palomares, Anna, Olvera-Maneu, Sergi, García-Calvo, Laura, López-Béjar, Manel, Martínez-Pastor, Felipe, and Álvarez-Rodríguez, Manuel
- Abstract
The effectiveness of rabbit-sperm cryopreservation is still below average compared to other domestic species. After the sperm cryopreservation process, post-thawing parameters like motility and membrane integrity are significantly compromised. The use of new extender constituents is an approach that can be used to improve the effectiveness of cryopreservation. Accordingly, we used honey (1.25, 2.5, 5, and 10%), coenzyme Q10 (100 and 200 μM), and β-carotene/α-tocopherol (500 μM/620 μM and 250 μM/310 μM) as candidate components for rabbit-sperm extenders during cryopreservation. Ejaculates from commercial adult rabbit bucks (n = 5) were cryopreserved using conventional freezing. Several post-thawing sperm parameters were assessed, including total motility, membrane integrity, viability, nuclear membrane integrity, acrosome reaction, and mitochondrial membrane potential and activation. Additionally, we performed hormonal analyses of the seminal plasma. Moreover, we analyzed the post-thawing levels of a molecular marker of sperm quality, proAKAP4, which was used in rabbits for the first time. Our findings showed that the 2.5% honey supplementation increased the post-thawing sperm motility (13.75 ± 3.75%) compared to the greater concentrations employed. However, the post-thawing motility was negatively affected by the coenzyme Q10 (0%, in both groups) but was not affected by the β-carotene/α-tocopherol supplementation (22 ± 18.15%, and 11.67 ± 10.17%). In conclusion, the cryopreservation protocols of this study did not help to maintain the sperm parameters after thawing. Further studies are required to identify novel protocols to mitigate the damage caused to rabbit sperm during cryopreservation.
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- 2023
49. The Cation/Calcium Channel of Sperm (CatSper): A Common Role Played Despite Inter-Species Variation?
- Author
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Vicente-Carrillo, Alejandro, Álvarez-Rodríguez, Manuel, Rodriguez-Martinez, Heriberto, Vicente-Carrillo, Alejandro, Álvarez-Rodríguez, Manuel, and Rodriguez-Martinez, Heriberto
- Abstract
The main cation/calcium channel of spermatozoa (CatSper), first identified in 2001, has been thoroughly studied to elucidate its composition and function, while its distribution among species and sperm sources is yet incomplete. CatSper is composed of several subunits that build a pore-forming calcium channel, mainly activated in vivo in ejaculated sperm cells by intracellular alkalinization and progesterone, as suggested by the in vitro examinations. The CatSper channel relevance is dual: to maintain sperm homeostasis (alongside the plethora of membrane channels present) as well as being involved in pre-fertilization events, such as sperm capacitation, hyperactivation of sperm motility and the acrosome reaction, with remarkable species differences. Interestingly, the observed variations in CatSper localization in the plasma membrane seem to depend on the source of the sperm cells explored (i.e., epididymal or ejaculated, immature or mature, processed or not), the method used for examination and, particularly, on the specificity of the antibodies employed. In addition, despite multiple findings showing the relevance of CatSper in fertilization, few studies have studied CatSper as a biomarker to fine-tune diagnosis of sub-fertility in livestock or even consider its potential to control fertilization in plague animals, a more ethically defensible strategy than implicating CatSper to pharmacologically modify male-related fertility control in humans, pets or wild animals. This review describes inter- and intra-species differences in the localization, structure and function of the CatSper channel, calling for caution when considering its potential manipulation for fertility control or improvement.
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- 2023
50. Refrigerated storage of ram sperm in presence of Trolox and GSH antioxidants: Effect of temperature, extender and storage time
- Author
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Mata-Campuzano, María, Álvarez-Rodríguez, Manuel, Tamayo-Canul, Julio, López-Urueña, Elena, de Paz, Paulino, Anel, Luis, Martínez-Pastor, Felipe, and Álvarez, Mercedes
- Published
- 2014
- Full Text
- View/download PDF
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