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Alvarez-Rodriguez, ManuelORCID iD iconorcid.org/0000-0003-0120-354X
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Publications (10 of 24) Show all publications
Martínez-Pastor, F., Álvarez, M., Guerra, C., Chamorro, C. A., Anel-López, L., de Paz, P., . . . Alvarez-Rodriguez, M. (2019). Extender osmolality, glycerol and egg yolk on the cryopreservation of epididymal spermatozoa for gamete banking of the Cantabric Chamois (Rupicapra pyrenaica parva). Theriogenology, 125, 109-114, Article ID S0093-691X(18)30573-9.
Open this publication in new window or tab >>Extender osmolality, glycerol and egg yolk on the cryopreservation of epididymal spermatozoa for gamete banking of the Cantabric Chamois (Rupicapra pyrenaica parva)
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2019 (English)In: Theriogenology, ISSN 0093-691X, E-ISSN 1879-3231, Vol. 125, p. 109-114, article id S0093-691X(18)30573-9Article in journal (Refereed) Published
Abstract [en]

Germplasm banking is a key technology enabling the ex-situ conservation of wild species. However, cryopreservation protocols must be tested to assure the applicability of the banked material. The objective of this study was defining a range of parameters for the composition of a semen extender for Cantabrian chamois epididymal spermatozoa (post-mortem collection). The freezing extender was based in a TES-Tris-fructose buffer, modifying its composition in three experiments: Osmolality of the buffer (320, 380 or 430 mOsm/kg, 8% glycerol, 15% egg yolk), glycerol (4 or 8%, 430 mOsm/kg, 15% egg yolk), egg yolk (10 or 15%, 430 mOsm/kg, 4% glycerol). Sperm was extended at 100 mill. spermatozoa/ml, cooled at 5 °C and frozen at -20 °C/min. Sperm quality was assessed pre and post-thawing (CASA, HOS test, abnormal forms, cytoplasmic droplets, and viability and acrosomal damage by flow cytometry). Freezability was good overall, with total motility of 65.5% ± 2.4 initial and 55.8% ± 2.4 post-thawing. The extenders affected the post-thaw sperm quality marginally. Whereas osmolalities and glycerol concentrations seemed not to differ, 430 mOsm/kg and 4% glycerol might be preferred. Egg yolk concentrations only differed on sperm velocity (VCL: 84.0 ± 6.7 μm/s in 10% vs. 70.7 ± 6.2 μm/s in 15%, P < 0.05). Our results suggest a good cryotolerance of chamois epididymal spermatozoa, with a preferred extender composition of hyperosmotic buffer, glycerol in the 4% range and lower egg yolk (10% range) than other ruminants.

Place, publisher, year, edition, pages
Elsevier, 2019
Keywords
Cantabrian chamois, Cryopreservation, Epididymal spermatozoa, Extender, Glycerol, Osmolality
National Category
Veterinary Science
Identifiers
urn:nbn:se:liu:diva-154015 (URN)10.1016/j.theriogenology.2018.10.022 (DOI)000455972500017 ()30408702 (PubMedID)2-s2.0-85055966888 (Scopus ID)
Available from: 2019-01-22 Created: 2019-01-22 Last updated: 2019-02-04Bibliographically approved
Atikuzzaman, M., Alvarez-Rodriguez, M., Carrillo, A. V., Johnsson, M., Wright, D. & Rodriguez-Martinez, H. (2017). Conserved gene expression in sperm reservoirs between birds and mammals in response to mating.. BMC Genomics, 18(1)
Open this publication in new window or tab >>Conserved gene expression in sperm reservoirs between birds and mammals in response to mating.
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2017 (English)In: BMC Genomics, ISSN 1471-2164, E-ISSN 1471-2164, Vol. 18, no 1Article in journal (Refereed) Published
Abstract [en]

BACKGROUND: Spermatozoa are stored in the oviductal functional sperm reservoir in animals with internal fertilization, including zoologically distant classes such as pigs or poultry. They are held fertile in the reservoir for times ranging from a couple of days (in pigs), to several weeks (in chickens), before they are gradually released to fertilize the newly ovulated eggs. It is currently unknown whether females from these species share conserved mechanisms to tolerate such a lengthy presence of immunologically-foreign spermatozoa. Therefore, global gene expression was assessed using cDNA microarrays on tissue collected from the avian utero-vaginal junction (UVJ), and the porcine utero-tubal junction (UTJ) to determine expression changes after mating (entire semen deposition) or in vivo cloacal/cervical infusion of sperm-free seminal fluid (SF)/seminal plasma (SP).

RESULTS: In chickens, mating changed the expression of 303 genes and SF-infusion changed the expression of 931 genes, as compared to controls, with 68 genes being common to both treatments. In pigs, mating or SP-infusion changed the expressions of 1,722 and 1,148 genes, respectively, as compared to controls, while 592 genes were common to both treatments. The differentially expressed genes were significantly enriched for GO categories related to immune system functions (35.72-fold enrichment). The top 200 differentially expressed genes of each treatment in each animal class were analysed for gene ontology. In both pig and chicken, an excess of genes affecting local immune defence were activated, though frequently these were down-regulated. Similar genes were found in both the chicken and pig, either involved in pH-regulation (SLC16A2, SLC4A9, SLC13A1, SLC35F1, ATP8B3, ATP13A3) or immune-modulation (IFIT5, IFI16, MMP27, ADAMTS3, MMP3, MMP12).

CONCLUSION: Despite being phylogenetically distant, chicken and pig appear to share some gene functions for the preservation of viable spermatozoa in the female reservoirs.

Place, publisher, year, edition, pages
BioMed Central, 2017
National Category
Biological Sciences
Identifiers
urn:nbn:se:liu:diva-134399 (URN)10.1186/s12864-017-3488-x (DOI)000394380200005 ()28100167 (PubMedID)
Note

Funding agencies: Research Council FORMAS, Stockholm [221-2011-512]

Available from: 2017-02-23 Created: 2017-02-23 Last updated: 2018-05-07
Najafi, A., Daghigh-Kia, H., Dodaran, H. V., Mehdipour, M. & Alvarez-Rodriguez, M. (2017). Ethylene glycol, but not DMSO, could replace glycerol inclusion in soybean lecithin-based extenders in ram sperm cryopreservation.. Animal Reproduction Science, 177, 35-41
Open this publication in new window or tab >>Ethylene glycol, but not DMSO, could replace glycerol inclusion in soybean lecithin-based extenders in ram sperm cryopreservation.
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2017 (English)In: Animal Reproduction Science, ISSN 0378-4320, E-ISSN 1873-2232, Vol. 177, p. 35-41Article in journal (Refereed) Published
Abstract [en]

The aim of this study was to evaluate the effects of glycerol, ethylene glycol or DMSO in a soybean lecithin extender for freezing ram semen. In this study, 20 ejaculates were collected from four Ghezel rams and diluted with soybean lecithin extender with glycerol (7%), ethylene glycol (3%, 5% and 7%) or DMSO (3%, 5% and 7%). Sperm motility (CASA), membrane integrity (HOS test), viability, total abnormality, mitochondrial activity (Rhodamine 123) and apoptotic features (Annexin V/Propidium iodide) were assessed after thawing. There was no significant difference between glycerol and ethylene glycol at different concentrations (3% and 5%) regarding sperm total and progressive motility, viability, and membrane integrity. The least percentages of mitochondrial functionality were observed in samples frozen with all different DMSO concentrations tested (P<0.05). Moreover, the percentage of post-thawed dead sperm was the greatest for all the DMSO concentrations compared with other groups (P<0.05). Thus, DMSO had an adverse effect on the post thaw ram sperm parameters. In contrast, ethylene glycol could be a desirable substitute of glycerol in the freezing extender, in view of similar results obtained in post-thaw quality of ram semen cryopreserved in a soybean lecithin extender. We propose that glycerol in a soybean lecithin based extender could be replaced by ethylene glycol at 3% or 5% concentrations.

Place, publisher, year, edition, pages
Elsevier, 2017
National Category
Biological Sciences
Identifiers
urn:nbn:se:liu:diva-134397 (URN)10.1016/j.anireprosci.2016.12.004 (DOI)000393267800004 ()28011116 (PubMedID)
Available from: 2017-02-23 Created: 2017-02-23 Last updated: 2018-05-02
Atikuzzaman, M., Sanz, L., Pla, D., Alvarez-Rodriguez, M., Rubér, M., Wright, D., . . . Rodriguez-Martinez, H. (2017). Selection for higher fertility reflects in the seminal fluid proteome of modern domestic chicken. Comparative Biochemistry and Physiology - Part D: Genomics and Proteomics, 21, 27-40
Open this publication in new window or tab >>Selection for higher fertility reflects in the seminal fluid proteome of modern domestic chicken
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2017 (English)In: Comparative Biochemistry and Physiology - Part D: Genomics and Proteomics, ISSN 1744-117X, E-ISSN 1878-0407, Vol. 21, p. 27-40Article in journal (Refereed) Published
Abstract [en]

The high egg-laying capacity of the modern domestic chicken (i.e. White Leghorn, WL) has arisen from the low egg-laying ancestor Red Junglefowl (RJF) via continuous trait selection and breeding. To investigate whether this long-term selection impacted the seminal fluid (SF)-proteome, 2DE electrophoresis-based proteomic analyses and immunoassays were conducted to map SF-proteins/cytokines in RJF, WL and a 9th generation Advanced Intercross Line (AIL) of RJF/WL-L13, including individual SF (n = 4, from each RJF, WL and AIL groups) and pools of the SF from 15 males of each group, analyzed by 2DE to determine their degree of intra-group (AIL, WL, and RJF) variability using Principal Component Analysis (PCA); respectively an inter-breed comparative analysis of intergroup fold change of specific SF protein spots intensity between breeds. The PCA clearly highlighted a clear intra-group similarity among individual roosters as well as a clear inter-group variability (e.g. between RJF, WL and AIL) validating the use of pools to minimize confounding individual variation. Protein expression varied considerably for processes related to sperm motility, nutrition, transport and survival in the female, including signaling towards immunomodulation. The major conserved SF-proteins were serum albumin and ovotransferrin. Aspartate aminotransferase, annexin A5, arginosuccinate synthase, glutathione S-transferase 2 and l-lactate dehydrogenase-A were RJF-specific. Glyceraldehyde-3-phosphate dehydrogenase appeared specific to the WL-SF while angiotensin-converting enzyme, γ-enolase, coagulation factor IX, fibrinogen α-chain, hemoglobin subunit α-D, lysozyme C, phosphoglycerate kinase, Src-substrate protein p85, tubulins and thioredoxin were AIL-specific. The RJF-SF contained fewer immune system process proteins and lower amounts of the anti-inflammatory/immunomodulatory TGF-β2 compared to WL and AIL, which had low levels- or lacked pro-inflammatory CXCL10 compared to RJF. The seminal fluid proteome differs between ancestor and modern chicken, with a clear enrichment of proteins and peptides related to immune-modulation for sperm survival in the female and fertility.

Place, publisher, year, edition, pages
Elsevier, 2017
Keywords
Rooster seminal fluid proteome, Cytokines, Egg-laying capacity, Red Junglefowl, White Leghorn, Advanced intercross line, Chicken
National Category
Biochemistry and Molecular Biology Medical Biotechnology (with a focus on Cell Biology (including Stem Cell Biology), Molecular Biology, Microbiology, Biochemistry or Biopharmacy) Genetics and Breeding
Identifiers
urn:nbn:se:liu:diva-132624 (URN)10.1016/j.cbd.2016.10.006 (DOI)000395224100004 ()27852008 (PubMedID)
Note

Funding agencies: Research Council FORMAS, Stockholm, Sweden [221-2011-512]; Ministerio de Ciencia e Innovacion (Madrid, Spain) [BFU2013-42833-P]

Available from: 2016-11-17 Created: 2016-11-17 Last updated: 2018-05-02Bibliographically approved
Fernández-Gago, R., Alvarez-Rodriguez, M., Alonso, M. E., González, J. R., Alegre, B., Domínguez, J. C. & Martínez-Pastor, F. (2017). Thawing boar semen in the presence of seminal plasma improves motility, modifies subpopulation patterns and reduces chromatin alterations.. Reproduction, fertility, and development, 29(8), 1576-1584
Open this publication in new window or tab >>Thawing boar semen in the presence of seminal plasma improves motility, modifies subpopulation patterns and reduces chromatin alterations.
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2017 (English)In: Reproduction, fertility, and development, ISSN 1031-3613, Vol. 29, no 8, p. 1576-1584Article in journal (Refereed) Published
Abstract [en]

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

Place, publisher, year, edition, pages
CSIRO PUBLISHING, 2017
National Category
Cell Biology
Identifiers
urn:nbn:se:liu:diva-130730 (URN)10.1071/RD15530 (DOI)000405782100012 ()27543989 (PubMedID)
Note

Funding agencies: Diputacion de Leon, Spain [DPI2009-08424]; Junta de Castilla y Leon, Spain; Ministry of Science and Innovation, Spain [RYC-2008-02560]

Available from: 2016-08-22 Created: 2016-08-22 Last updated: 2018-05-02
Alvarez-Rodriguez, M., Alvarez, M., Anel-Lopez, L., Lopez-Uruena, E., Manrique, P., Borragan, S., . . . Anel, L. (2016). Effect of colloid (Androcoll-Bear, Percoll, and PureSperm) selection on the freezability of brown bear (Ursus arctos) sperm. Theriogenology, 85(6), 1097-1105
Open this publication in new window or tab >>Effect of colloid (Androcoll-Bear, Percoll, and PureSperm) selection on the freezability of brown bear (Ursus arctos) sperm
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2016 (English)In: Theriogenology, ISSN 0093-691X, E-ISSN 1879-3231, Vol. 85, no 6, p. 1097-1105Article in journal (Refereed) Published
Abstract [en]

The development of a species-specific conservation protocol that involves artificial insemination with frozen semen needs to validate an effective methodology for freezing semen. Colloid centrifugation has been suggested and widely applied as an effective tool for selecting animal spermatozoa for artificial breeding. The objective of the present study was to compare different methods of centrifugation, single layer using Androcoll-Bear and Percoll and double layer using PureSperm 100 (in two different discontinuous gradients 40%-80% and 45%-90%), for the selection of fresh brown bear sperm samples. In the before freezing group, all selected samples showed a higher progressive motility and viability (except Percoll for motility 43.0 +/- 5.3 [P &lt; 0.051); all colloids except PureSperm 45/90% rendered samples with fewer damaged acrosomes. In the after thawing group, all tested centrifugation colloids showed a good capacity to decrease the number of damaged acrosomes. Furthermore, PureSperm treatment (45/90%) resulted in an increase in apoptotic-like changes not only immediately after thawing but also after the incubation test, leading us to suggest that this gradient could induce some kind of deleterious effects on the sperm samples. On the other hand, PureSperm treatment (40/80%) yielded a quality preservation capacity similar to Androcoll-Bear in number of damaged acrosomes, different relative to the control (control, 5.3 +/- 0.6; PureSperm 80, 2.0 +/- 0.3; Androcoll, 2.1 +/- 0.9 [P &lt; 0.051) but a decrease in the number of viable spermatozoa recovered after thawing relative to the control (control, 21.2 +/- 3.1; PureSperm 80, 13.7 +/- 2.7 [P &lt; 0.051). In conclusion, Androcoll-Bear constitutes a useful tool for handling of brown bear ejaculates owing to its simple handling and procedure with a reliable sperm selection and freezability. This colloid yielded an improvement in several sperm parameters in brown bear frozen-thawed semen; the selected spermatozoa of fresh samples with this colloid showed a better resistance to freezing compared with the control sample not only for motility but also for viability. (C) 2016 Elsevier Inc. All rights reserved.

Place, publisher, year, edition, pages
ELSEVIER SCIENCE INC, 2016
Keywords
Brown bear sperm; Androcoll; PureSperm; Percoll; Freezability
National Category
Clinical Medicine
Identifiers
urn:nbn:se:liu:diva-126803 (URN)10.1016/j.theriogenology.2015.11.021 (DOI)000371453500010 ()26764151 (PubMedID)
Note

Funding Agencies|MINECO [CGL2013-48255-R]; CANTUR, S.A.

Available from: 2016-04-07 Created: 2016-04-05 Last updated: 2017-11-30
Vicente-Carrillo, A., Ekwall, H., Álvarez-Rodríguez, M. & Rodriguez-Martinez, H. (2016). Membrane Stress During Thawing Elicits Redistribution of Aquaporin 7 But Not of Aquaporin 9 in Boar Spermatozoa. Reproduction in domestic animals, 51(5), 665-679
Open this publication in new window or tab >>Membrane Stress During Thawing Elicits Redistribution of Aquaporin 7 But Not of Aquaporin 9 in Boar Spermatozoa
2016 (English)In: Reproduction in domestic animals, ISSN 0936-6768, E-ISSN 1439-0531, Vol. 51, no 5, p. 665-679Article in journal (Refereed) Published
Abstract [en]

Freezing of boar spermatozoa includes the cryoprotectant glycerol, but renders low cryosurvival, owing to major changes in osmolarity during freezing/thawing. We hypothesize that aquaporins (AQPs) 7 and 9 adapt their membrane domain location to these osmotic challenges, thus maintaining sperm homeostasis. Western blotting (WB) and immunocytochemistry (ICC) at light and electron microscope levels with several commercial primary antibodies and protocols explored AQP location on cauda epididymal and ejaculated spermatozoa (from different fractions of the ejaculate), unprocessed, extended, chilled and frozen-thawed. Although differences in WB and ICC labelling were seen among antibodies, AQP-7 was conspicuously located in the entire tail and cytoplasmic droplet in caudal spermatozoa, being restricted to the mid-piece and principal piece domains in ejaculated spermatozoa. AQP-9 was mainly localized in the sperm head in both caudal and ejaculated spermatozoa. While unaffected by chilling (+5°C), freezing and thawing of ejaculated spermatozoa clearly relocated the head labelling of AQP-7, but not that of AQP-9. In vitro mimicking of cell membrane expansion during quick thawing maintained the localization of AQP-9 but relocated AQP-7 towards the acrosome. AQP-7, but not AQP-9, appears as a relevant marker for non-empirical studies of sperm handling.

Place, publisher, year, edition, pages
Blackwell Verlag, 2016
National Category
Developmental Biology
Identifiers
urn:nbn:se:liu:diva-130228 (URN)10.1111/rda.12728 (DOI)000388334100005 ()27405395 (PubMedID)
Note

Funding agencies: Swedish Research Council VR, Stockholm [521-2011-6553]; Research Council FORMAS, Stockholm [221-2011-512]; FORSS (Forskningsradet i Sydostra Sverige), Sweden [473121]

Available from: 2016-07-19 Created: 2016-07-19 Last updated: 2018-03-23Bibliographically approved
Vicente Carrillo, A., Alvarez-Rodriguez, M. & Rodriguez-Martinez, H. (2016). The mu (µ) and delta (δ) opioid receptors modulate boar sperm motility. Molecular Reproduction and Development, 83(8), 724-734
Open this publication in new window or tab >>The mu (µ) and delta (δ) opioid receptors modulate boar sperm motility
2016 (English)In: Molecular Reproduction and Development, ISSN 1040-452X, E-ISSN 1098-2795, Vol. 83, no 8, p. 724-734Article in journal (Refereed) Published
Abstract [en]

Endogenous and exogenous opioids modulate reproductive functions in target cells via opioid receptors (µ, δ, and κ). Sperm motility is a metric of gamete functionality, and serves as a suitable parameter for in vitro drug-induced toxicity assays. This study identifies the presence and location of opioid receptors in pig spermatozoa as well as their functional response after in vitro challenge with known agonists (morphine [µ]; [D-Pen 2,5]-enkephanile [δ]; and U 50488 [κ]) and antagonists (naloxone [µ]; naltrindole [δ]; and nor-binaltrorphimine [κ]). Only the µ- and δ-opioid receptors were present in the sperm plasma membrane, overlying the acrosome, neck, and principal piece. Challenge experiments with agonists and antagonists identified both µ- and δ-opioid receptors as regulators of sperm kinematics, wherein µ maintains or increases sperm movement whereas δ decreases sperm motility over time. This article is protected by copyright. All rights reserved.

Place, publisher, year, edition, pages
John Wiley & Sons, 2016
Keywords
opioids, membrane receptors, kinematics, pig
National Category
Developmental Biology
Identifiers
urn:nbn:se:liu:diva-130181 (URN)10.1002/mrd.22675 (DOI)000387014800007 ()27391529 (PubMedID)
Note

Funding agencies: Swedish Research Council VR, Stockholm [521-2011-6553]; Research Council FORMAS, Sweden [221-2011-512]; Research Council in Southeast Sweden (FORSS), Sweden [378091/31297]

Available from: 2016-07-14 Created: 2016-07-14 Last updated: 2017-11-28Bibliographically approved
Rodriguez-Martinez, H., Tienthai, P., Atikuzzaman, M., Vicente Carrillo, A., Rubér, M. & Alvarez-Rodriguez, M. (2016). The ubiquitous hyaluronan: Functionally implicated in the oviduct?. Theriogenology, 86(1), 182-186
Open this publication in new window or tab >>The ubiquitous hyaluronan: Functionally implicated in the oviduct?
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2016 (English)In: Theriogenology, ISSN 0093-691X, E-ISSN 1879-3231, Vol. 86, no 1, p. 182-186Article, review/survey (Refereed) Published
Abstract [en]

Hyaluronan (hyaluronic acid) is a simple, nonantigenic, nonsulfated glycosaminoglycan present everywhere in the extracellular compartments of the body. Noteworthy, it is highly conserved phylogenetically, from sauropsida to mammals; and plays a plethora of roles from embryonic/fetal development to adult physiological and pathological events, including tumor development. In reproduction, hyaluronan has proven related to initial events as sperm survival, buildup of the sperm reservoir in the oviduct, regulation of sperm capacitation, and prefertilization to later participate in embryo, fetal, and placental development. Synthesis, binding (via the CD44 membrane receptor), and degradation of hyaluronan occur in male and female genital organs, the oviduct being no exception. This review discusses our current knowledge on roles of this ubiquitous glycosaminoglycan on the survival of immunologically foreign spermatozoa in the pig oviduct, a relevant event for fertility. During preovulatory storage in the functional tubal sperm reservoir, spermatozoa are entrapped in a mucus-like tubal fluid. This fluid contains fluctuating levels of hyaluronan, which is synthesized by the lining epithelium by hyaluronan synthase 3. Both hyaluronan and its CD44 receptor are particularly evident in the deep mucosal furrows of the sperm reservoir, in which most spermatozoa are embedded in; kept alive, uncapacitated but also undetected by the immune system of the female. Hyaluronan is also present in the seminal plasma, and evidence points toward an involvement of hyaluronan and its receptor in the local (tubal and possibly uterine) production of antiinflammatory cytokines, such as interleulcin-10, pertaining maternal immune tolerance of these foreign cells. (C) 2015 Elsevier Inc. All rights reserved.

Place, publisher, year, edition, pages
ELSEVIER SCIENCE INC, 2016
Keywords
Hyaluronan; Sperm oviduct reservoir; Sperm survival; Capacitation; Immunity; Pig
National Category
Obstetrics, Gynecology and Reproductive Medicine
Identifiers
urn:nbn:se:liu:diva-130123 (URN)10.1016/j.theriogenology.2015.11.025 (DOI)000377643700020 ()26768539 (PubMedID)
Note

Funding Agencies|Swedish Research Council VR (Vetenskapsradet) [521-2011-6353]; Swedish Research Council Formas, Stockholm, Sweden [221-2011-512]

Available from: 2016-07-12 Created: 2016-07-11 Last updated: 2017-11-28
Mata-Campuzano, M., Álvarez-Rodríguez, M., Álvarez, M., Tamayo-Canul, J., Anel, L., de Paz, P. & Martínez-Pastor, F. (2015). Post-thawing quality and incubation resilience of cryopreserved ram spermatozoa are affected by antioxidant supplementation and choice of extender.. Theriogenology, 83(4), 520-528
Open this publication in new window or tab >>Post-thawing quality and incubation resilience of cryopreserved ram spermatozoa are affected by antioxidant supplementation and choice of extender.
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2015 (English)In: Theriogenology, ISSN 0093-691X, E-ISSN 1879-3231, Vol. 83, no 4, p. 520-528Article in journal (Refereed) Published
Abstract [en]

The performance of cryopreserved semen in ovine artificial insemination still needs improvement. Some antioxidants have been tested, with variable success. We cryopreserved semen from Churra rams using TES-Tris-fructose with 4% glycerol and 10% egg yolk (EY) or 3.5% soybean lecithin (SL), with 1 mM of reduced glutathione (GSH), Trolox, crocin, or cysteamine. Samples were analyzed after thawing and incubation (6 hours, 38 °C) for motility (computer-assisted sperm analysis [CASA]), viability, acrosomal integrity, apoptosis, mitochondrial activity, chromatin status, and lipoperoxidation (malondialdehyde production). Interactions (antioxidant/extender/incubation) were significant for most variables. Extenders yielded similar results, although SL depressed mitochondrial activity and linearity (P < 0.001), it improved motility (P < 0.05), DNA fragmentation (P < 0.05), and acrosomal damage (P < 0.001). The control, GSH, and Trolox showed greater viability with SL (P < 0.01). Cysteamine depressed motility (0 hours: 51.6 ± 2.0% vs. 32.3 ± 4.3%; 6 hours: no motility vs. 32.5 ± 1.9%; P < 0.001), but improved viability when using EY (P = 0.004). Crocin increased acrosomal damage (P = 0.022) but improved linearity-related parameters after thawing (P = 0.014). Trolox considerably reduced malondialdehyde production in both extenders (8.6 ± 0.4 nmol per 10(8) cells vs. 14.2 ± 0.3 in EY and 20 ± 0.6 in SL; P < 0.001). Interestingly, thiol antioxidants (cysteamine and GSH) increased DNA fragmentation (percentage of DNA fragmentation index), whereas crocin reduced it (P < 0.05). Interactions between extender and antioxidant must be taken into account for improving sperm cryopreservation. Soybean lecithin seems to be a suitable replacement for EY, but its effect on mitochondria must be investigated. Trolox and crocin might be useful for ram semen freezing.

Place, publisher, year, edition, pages
Elsevier, 2015
Keywords
Ram, Spermatozoa, Antioxidant, Cryopreservation, Extender
National Category
Developmental Biology
Identifiers
urn:nbn:se:liu:diva-130184 (URN)10.1016/j.theriogenology.2014.10.018 (DOI)25499089 (PubMedID)
Available from: 2016-07-14 Created: 2016-07-14 Last updated: 2017-11-28Bibliographically approved
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ORCID iD: ORCID iD iconorcid.org/0000-0003-0120-354X

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