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  1. Article ; Online: Heat SHOCK proteins in equine spermatozoa: Expression and correlation to kinetic and environmental parameters.

    Albrizio, M / Lacalandra, G M / Volpe, S / Nicassio, M / Cinone, M

    Theriogenology

    2020  Volume 155, Page(s) 185–196

    Abstract: Heat Shock Proteins are chaperones primary involved in the repair of cellular damages induced by temperature. The harmful effect of temperature on the male gonad is well known, on the contrary knowledge on the effects of the environment on semen quality ... ...

    Abstract Heat Shock Proteins are chaperones primary involved in the repair of cellular damages induced by temperature. The harmful effect of temperature on the male gonad is well known, on the contrary knowledge on the effects of the environment on semen quality are still insufficient. The aim of this paper was to learn more about the role of HSPs and the environment in modulating the physiology of equine male gonads. We showed a detailed analysis of equine semen characteristic and the expression level of three HSPs (60-70-90) over a one-year period analyzing the effects of temperature and humidity and the correlation among the different variables. We showed also that the interpretation of results depends strongly on the way in which data are assembled and analyzed, therefore we compared results obtained from three different ways of grouping: according to single months, to weather seasons and to mare reproductive periods. Results showed that the expression of the three HSPs is correlated to the environment through temperature and humidity and that it reaches the highest level in the breeding season and in summer. We found also that HSPs expression is correlated to some variables describing the quality of equine semen (concentration) and the kinetic of spermatozoa (total motility-MOT, %, average path velocity -VAP, μm/s- and lateral head displacement -ALH, μm). No correlation was found between HSPs expression and the mitochondrial membrane potential; while viability and HSP90 expression resulted positively correlated. The month-by-month analysis evidenced that in February equine semen has the highest kinetic characteristics (increased linearity -LIN, %-, straightness -STR, % -and average path velocity -VAP, μm/s) with the highest number of motile, progressive motile and rapid cells. These results may have a great impact in the comprehension of functional aspects of the physiology of equine semen and may have potential implications for breeders who want to understand the period (and/or month) of the year in which equine semen reaches the best characteristics with increased chances for better results in reproductive practice.
    MeSH term(s) Animals ; Female ; Heat-Shock Proteins/genetics ; Horses ; Male ; Semen ; Semen Analysis/veterinary ; Sperm Motility ; Spermatozoa
    Chemical Substances Heat-Shock Proteins
    Language English
    Publishing date 2020-06-06
    Publishing country United States
    Document type Journal Article
    ZDB-ID 189232-0
    ISSN 1879-3231 ; 0093-691X
    ISSN (online) 1879-3231
    ISSN 0093-691X
    DOI 10.1016/j.theriogenology.2020.05.042
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  2. Article: Heat SHOCK proteins in equine spermatozoa: Expression and correlation to kinetic and environmental parameters

    Albrizio, M / Lacalandra, G.M / Volpe, S / Nicassio, M / Cinone, M

    Theriogenology. 2020 Oct. 01, v. 155

    2020  

    Abstract: Heat Shock Proteins are chaperones primary involved in the repair of cellular damages induced by temperature. The harmful effect of temperature on the male gonad is well known, on the contrary knowledge on the effects of the environment on semen quality ... ...

    Abstract Heat Shock Proteins are chaperones primary involved in the repair of cellular damages induced by temperature. The harmful effect of temperature on the male gonad is well known, on the contrary knowledge on the effects of the environment on semen quality are still insufficient. The aim of this paper was to learn more about the role of HSPs and the environment in modulating the physiology of equine male gonads. We showed a detailed analysis of equine semen characteristic and the expression level of three HSPs (60-70-90) over a one-year period analyzing the effects of temperature and humidity and the correlation among the different variables. We showed also that the interpretation of results depends strongly on the way in which data are assembled and analyzed, therefore we compared results obtained from three different ways of grouping: according to single months, to weather seasons and to mare reproductive periods. Results showed that the expression of the three HSPs is correlated to the environment through temperature and humidity and that it reaches the highest level in the breeding season and in summer. We found also that HSPs expression is correlated to some variables describing the quality of equine semen (concentration) and the kinetic of spermatozoa (total motility-MOT, %, average path velocity -VAP, μm/s- and lateral head displacement -ALH, μm). No correlation was found between HSPs expression and the mitochondrial membrane potential; while viability and HSP90 expression resulted positively correlated. The month-by-month analysis evidenced that in February equine semen has the highest kinetic characteristics (increased linearity –LIN, %-, straightness –STR, % -and average path velocity -VAP, μm/s) with the highest number of motile, progressive motile and rapid cells.These results may have a great impact in the comprehension of functional aspects of the physiology of equine semen and may have potential implications for breeders who want to understand the period (and/or month) of the year in which equine semen reaches the best characteristics with increased chances for better results in reproductive practice.
    Keywords head ; heat stress ; humidity ; males ; mares ; membrane potential ; mitochondrial membrane ; semen quality ; summer ; temperature ; viability
    Language English
    Dates of publication 2020-1001
    Size p. 185-196.
    Publishing place Elsevier Inc.
    Document type Article
    Note NAL-AP-2-clean
    ZDB-ID 189232-0
    ISSN 1879-3231 ; 0093-691X
    ISSN (online) 1879-3231
    ISSN 0093-691X
    DOI 10.1016/j.theriogenology.2020.05.042
    Database NAL-Catalogue (AGRICOLA)

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  3. Article ; Online: Localization and functional modification of L-type voltage-gated calcium channels in equine spermatozoa from fresh and frozen semen.

    Albrizio, M / Moramarco, A M / Nicassio, M / Micera, E / Zarrilli, A / Lacalandra, G M

    Theriogenology

    2015  Volume 83, Issue 3, Page(s) 421–429

    Abstract: It is well known that insemination of cryopreserved semen always results in lower fertility when compared with fresh semen, but there is an increased interest and demand for frozen equine semen by the major breeder associations because of the utility ... ...

    Abstract It is well known that insemination of cryopreserved semen always results in lower fertility when compared with fresh semen, but there is an increased interest and demand for frozen equine semen by the major breeder associations because of the utility arising from semen already "on hand" at breeding time. In this article, we report that equine sperm cells express L-type voltage-gated calcium channels; their localization is restricted to sperm neck and to the principal piece of the tail in both fresh and frozen-thawed spermatozoa. We also studied the causes of cryoinjury at the membrane level focusing on the function of L-type calcium channels. We report that in cryopreserved spermatozoa the mean basal value of [Ca(2+)]i is higher than that of spermatozoa from fresh semen (447.130 vs. 288.3 nM; P < 0.001) and L-type channels function differently in response to their agonist and antagonist in relation to semen condition (fresh or frozen-thawed). We found that on addition of agonist to the culture medium, the increase in intracellular calcium concentrations ([Ca(2+)]i) was greater in frozen semen than in fresh semen (Δ[Ca(2+)]i = 124.59 vs. 16.04 nM; P < 0.001), whereas after the addition of antagonist the decrease in [Ca(2+)]i was lower in frozen semen than in fresh semen (Δ[Ca(2+)]i = 32.5 vs. 82.5 nM; P < 0.001). In this article, we also discuss the impact of cryopreservation on sperm physiology.
    MeSH term(s) Animals ; Calcium/metabolism ; Calcium Channels, L-Type/analysis ; Calcium Channels, L-Type/chemistry ; Calcium Channels, L-Type/metabolism ; Cryopreservation/veterinary ; Horses/metabolism ; Male ; Semen Preservation/adverse effects ; Semen Preservation/veterinary ; Spermatozoa/metabolism ; Spermatozoa/physiology
    Chemical Substances Calcium Channels, L-Type ; Calcium (SY7Q814VUP)
    Language English
    Publishing date 2015-02
    Publishing country United States
    Document type Comparative Study ; Journal Article
    ZDB-ID 189232-0
    ISSN 1879-3231 ; 0093-691X
    ISSN (online) 1879-3231
    ISSN 0093-691X
    DOI 10.1016/j.theriogenology.2014.10.005
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  4. Article ; Online: Effects of in vitro exposure to natural levels of zearalenone and its derivatives on chromatin structure stability in equine spermatozoa.

    Minervini, F / Lacalandra, G M / Filannino, A / Nicassio, M / Visconti, A / Dell'Aquila, M E

    Theriogenology

    2010  Volume 73, Issue 3, Page(s) 392–403

    Abstract: The purpose of this study was to assess the natural exposure of male horses (Equus caballus) to the mycotoxin zearalenone (ZEA) by using the ELISA test and to evaluate the effects of in vitro exposure of sperm cells to mycotoxin-containing urine extracts ...

    Abstract The purpose of this study was to assess the natural exposure of male horses (Equus caballus) to the mycotoxin zearalenone (ZEA) by using the ELISA test and to evaluate the effects of in vitro exposure of sperm cells to mycotoxin-containing urine extracts on sperm chromatin structure stability. Because of their occurrence in urine samples, ZEA and its derivatives were tested by sperm chromatin structure assay (SCSA) at natural levels detected by ELISA. Thirty-eight urine extracts of Italian (n = 11) and northeastern European (n = 27) horses were tested on frozen-thawed spermatozoa to evaluate the toxic effect of mycotoxin on their chromatin structure by flow cytometry. Different parameters of the DNA fragmentation index (DFI), such as the mean (X -DFI), the percentage (%-DFI), and the standard deviation (SD-DFI), were analyzed. Urine samples showed a mean level of 32.3 ng/mL ZEA with significantly higher concentrations in northeastern European samples than in Italian samples, probably in relation to climatic and feeding differences. The toxic effects of ZEA-containing urine samples on SCSA parameters were found at low ZEA concentrations and were mainly observed in Italian samples. By using mycotoxin standards, ZEA, alpha-zearalenol, and beta-zearalenol proved to be more toxic compounds for sperm chromatin stability than other tested derivatives. A nongenomic mechanism of action can be hypothesized.
    MeSH term(s) Animals ; Chromatin/drug effects ; Chromatin/ultrastructure ; Enzyme-Linked Immunosorbent Assay ; Estrogens, Non-Steroidal/toxicity ; Horses/genetics ; Male ; Spermatozoa/drug effects ; Zearalenone/toxicity ; Zearalenone/urine
    Chemical Substances Chromatin ; Estrogens, Non-Steroidal ; Zearalenone (5W827M159J)
    Language English
    Publishing date 2010-02
    Publishing country United States
    Document type Journal Article
    ZDB-ID 189232-0
    ISSN 1879-3231 ; 0093-691X
    ISSN (online) 1879-3231
    ISSN 0093-691X
    DOI 10.1016/j.theriogenology.2009.09.023
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  5. Article ; Online: First evidence of placental transfer of ochratoxin A in horses.

    Minervini, Fiorenza / Giannoccaro, Alessandra / Nicassio, Michele / Panzarini, Giuseppe / Lacalandra, Giovanni Michele

    Toxins

    2013  Volume 5, Issue 1, Page(s) 84–92

    Abstract: Ochratoxin A (OTA) is a renal mycotoxin and transplacental genotoxic carcinogen. The aim of this study was to evaluate the natural occurrence of OTA in equine blood samples and its placental transfer. For the assessment of OTA levels, serum samples were ... ...

    Abstract Ochratoxin A (OTA) is a renal mycotoxin and transplacental genotoxic carcinogen. The aim of this study was to evaluate the natural occurrence of OTA in equine blood samples and its placental transfer. For the assessment of OTA levels, serum samples were collected from 12 stallions, 7 cycling mares and 17 pregnant mares. OTA was found in 83% of serum samples (median value = 121.4 pg/mL). For the assessment of placental transfer, serum samples were collected from the 17 mares after delivery and from the umbilical cords of their foals, after foaling. Fourteen serum samples from pregnant mares contained OTA (median value = 106.5 pg/mL), but only 50% of their foals were exposed (median values = 96.6 pg/mL). HPLC analysis carried out on four serum samples (collected from two mares and their respective foals) supported the ELISA results on OTA placental transfer. This is the first report on the natural occurrence of OTA in horse serum samples and placental transfer in horses.
    MeSH term(s) Animals ; Animals, Newborn/blood ; Animals, Newborn/metabolism ; Carcinogens/pharmacokinetics ; Chromatography, High Pressure Liquid ; Enzyme-Linked Immunosorbent Assay ; Female ; Fetal Blood/chemistry ; Horses/physiology ; Male ; Maternal-Fetal Exchange ; Mycotoxins/blood ; Mycotoxins/pharmacokinetics ; Ochratoxins/blood ; Ochratoxins/pharmacokinetics ; Placenta/metabolism ; Pregnancy
    Chemical Substances Carcinogens ; Mycotoxins ; Ochratoxins ; ochratoxin A (1779SX6LUY)
    Language English
    Publishing date 2013-01-11
    Publishing country Switzerland
    Document type Journal Article
    ZDB-ID 2518395-3
    ISSN 2072-6651 ; 2072-6651
    ISSN (online) 2072-6651
    ISSN 2072-6651
    DOI 10.3390/toxins5010084
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  6. Article ; Online: Lectin-binding sites on ejaculated stallion sperm during breeding and non-breeding periods.

    Desantis, S / Ventriglia, G / Zizza, S / Nicassio, M / Valentini, L / Di Summa, A / Lacalandra, G M

    Theriogenology

    2010  Volume 73, Issue 8, Page(s) 1146–1153

    Abstract: Stallion sperm from semen collected in Southern Italy during the breeding (June-July) and non-breeding (December-January) periods were analyzed by means of twelve lectins to evaluate the glycoconjugate pattern and to verify whether there are any seasonal ...

    Abstract Stallion sperm from semen collected in Southern Italy during the breeding (June-July) and non-breeding (December-January) periods were analyzed by means of twelve lectins to evaluate the glycoconjugate pattern and to verify whether there are any seasonal differences in the glycosylation pattern of the sperm glycocalyx. The acrosomal cap showed reactivity for Maackia amurensis (MAL II), Sambucus nigra (SNA), Arachis hypogaea (PNA), Glycine max (SBA), Helix pomatia (HPA), Canavalia ensiformis (Con A) Triticum vulgaris (WGA), and Griffonia simplicifolia isolectin II (GSA II) in breeding and non-breeding ejaculated sperm, suggesting the presence of oligosaccharides terminating with Neu5Ac alpha 2,3Gal beta 1,4GlcNAc, Neu5Ac alpha 2,6Gal/GalNAc, with Gal beta 1,3GalNAc, alpha/beta GalNAc and glycans with terminal/internal alpha Man and GlcNAc. During the non-breeding period, the acrosomal cap expressed oligosaccharides terminating with Gal beta 1,4GlcNAc (Ricinus communis(120) affinity) (RCA(120)) and L-Fuc alpha 1,2Gal beta 1,4GlcNAc beta (Ulex europaeus affinity) (UEA I). The equatorial segment placed between the acrosomal cap and post-acrosomal region did not display glycans terminating with GalNAc, GlcNAc, and alpha L-Fuc. The post-acrosomal region of sperm collected in the breeding and non-breeding periods bound Con A, MAL II, SNA, and SBA, thus showing the presence of N-linked oligosaccharides from high-Man content, terminating with Neu5Ac alpha 2,3Gal beta 1,4GlcNAc, Neu5Ac alpha 2,6Gal/GalNAc and GalNAc. In winter, the post-acrosomal region also expressed oligosaccharides terminating with alpha GalNAc, GlcNAc, and L-Fuc alpha 1,2Gal beta 1,4GlcNAc beta (HPA, GSA II, and UEA I staining). The tail of sperm from semen collected during the breeding and non-breeding periods showed a lectin binding pattern similar to the post-acrosomal region, except for the absence of HPA staining in sperm collected during the winter season. These results indicate that the surface of stallion sperm contains different glycocalyx domains and that the glycosylation pattern undergoes changes during the breeding and non-breeding periods.
    MeSH term(s) Animals ; Binding Sites ; Ejaculation ; Glycosylation ; Horses/metabolism ; Horses/physiology ; Lectins/chemistry ; Lectins/metabolism ; Male ; Protein Binding ; Reproduction/physiology ; Semen Analysis ; Sexual Behavior, Animal/physiology ; Sperm Retrieval/veterinary ; Spermatozoa/chemistry ; Spermatozoa/metabolism ; Time Factors ; Tissue Distribution
    Chemical Substances Lectins
    Language English
    Publishing date 2010-05
    Publishing country United States
    Document type Comparative Study ; Journal Article ; Research Support, Non-U.S. Gov't
    ZDB-ID 189232-0
    ISSN 1879-3231 ; 0093-691X
    ISSN (online) 1879-3231
    ISSN 0093-691X
    DOI 10.1016/j.theriogenology.2009.12.001
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  7. Article: In vitro equine oocyte maturation in pure follicular fluid plus interleukin-1 and fertilization following ICSI.

    Caillaud, M / Dell'aquila, M E / De Santis, T / Nicassio, M / Lacalandra, G M / Goudet, G / Gérard, N

    Animal reproduction science

    2008  Volume 106, Issue 3-4, Page(s) 431–439

    Abstract: The interleukin-1 (IL-1) system is thought to be involved in periovulatory events in the mare. Previous in vivo studies have demonstrated that IL-1beta induces oocyte maturation, but depresses the pregnancy rate 14 days after ovulation. To better ... ...

    Abstract The interleukin-1 (IL-1) system is thought to be involved in periovulatory events in the mare. Previous in vivo studies have demonstrated that IL-1beta induces oocyte maturation, but depresses the pregnancy rate 14 days after ovulation. To better understand the role of IL-1 in oocyte maturation and fertilization, the effects of IL-1 on the in vitro maturation rate of equine oocytes in pure follicular fluid were evaluated and fertilization rate assessed following intracytoplasmic sperm injection (ICSI). Oocytes collected from slaughterhouse ovaries were cultured in four different media for 30 h prior to fertilization. Two experiments were performed, each using three maturation media as the experimental treatments. Medium 1 was pure follicular fluid from subordinate follicles. Medium 2 was medium 1 plus 50 ng/ml recombinant human IL-1beta. Medium 3 was pure follicular fluid collected from mares administered crude equine gonadotropin (CEG). Medium 4 was medium 2 plus 50 ng/ml of recombinant human IL-1 receptor antagonist. Media 1, 2 and 3 were compared in experiment 1. In experiment 2, media 1, 2 and 4 were compared. After maturation, metaphase II oocytes were submitted to microinjection and assessed for signs of fertilization. In experiment 1, 101 oocytes were evaluated. The rate of polar body extrusion was 66, 51 and 68% and the proportions of normally fertilized oocytes after ICSI were 40, 18 and 38% for media 1, 2 and 3, respectively. In experiment 2, 122 oocytes were evaluated. The rate of polar body extrusion was 55, 48 and 42% and the proportions showing normal fertilization after ICSI were 14, 25 and 29% for media 1, 2 and 4, respectively. There was no positive effect of IL-1beta on maturation in both experiments, but the fertilization rate and percentage of embryos reaching four-cell were low in the presence of IL-1beta, indicating that this cytokine may interfere with fertilization and early embryo development.
    MeSH term(s) Animals ; Cells, Cultured ; Culture Media/pharmacology ; Embryo, Mammalian/cytology ; Female ; Fertilization/drug effects ; Follicular Fluid/physiology ; Horses/physiology ; Interleukin-1beta/pharmacology ; Oocytes/drug effects ; Oocytes/physiology ; Oogenesis/drug effects ; Pregnancy ; Pregnancy Rate ; Pregnancy, Animal ; Sperm Injections, Intracytoplasmic/drug effects
    Chemical Substances Culture Media ; Interleukin-1beta
    Language English
    Publishing date 2008-07
    Publishing country Netherlands
    Document type Evaluation Studies ; Journal Article ; Research Support, Non-U.S. Gov't
    ZDB-ID 429674-6
    ISSN 1873-2232 ; 0378-4320
    ISSN (online) 1873-2232
    ISSN 0378-4320
    DOI 10.1016/j.anireprosci.2007.06.005
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  8. Article ; Online: Exposure to follicular fluid during oocyte maturation and oviductal fluid during post-maturation does not improve in vitro embryo production in the horse.

    Douet, Cécile / Parodi, Olivia / Martino, Nicola Antonio / Lacalandra, Giovanni Michele / Nicassio, Michele / Reigner, Fabrice / Deleuze, Stefan / Dell'Aquila, Maria Elena / Goudet, Ghylène

    Zygote (Cambridge, England)

    2017  Volume 25, Issue 5, Page(s) 612–630

    Abstract: Most wild equids and many domestic horse breeds are at risk of extinction, so there is an urgent need for genome resource banking. Embryos cryopreservation allows the preservation of genetics from male and female and is the fastest method to restore a ... ...

    Abstract Most wild equids and many domestic horse breeds are at risk of extinction, so there is an urgent need for genome resource banking. Embryos cryopreservation allows the preservation of genetics from male and female and is the fastest method to restore a breed. In the equine, embryo production in vitro would allow the production of several embryos per cycle. Intracytoplasmic sperm injection (ICSI) is used to generate horse embryos, but it requires expensive equipment and expertise in micromanipulation, and blastocyst development rates remain low. No conventional in vitro fertilization (IVF) technique for equine embryo production is available. The development of culture conditions able to mimic the maturation of the oocyte in preovulatory follicular fluid (pFF) and the post-maturation in oviductal fluid (OF) may improve embryo production in vitro. Our aim was to analyse the effect of in vitro maturation in pFF and incubation in OF on in vitro maturation of equine oocytes, fertilization using conventional IVF or ICSI, and embryo development after culture in synthetic oviductal fluid (SOF) or DMEM-F12. Oocytes collected from slaughtered mares or by ovum pick up were matured in vitro in pFF or semi-synthetic maturation medium (MM). The in vitro maturation, fertilization and development rates were not statistically different between pFF and MM. After in vitro maturation, oocytes were incubated with or without OF. Post-maturation in OF did not significantly improve the fertilization and development rates. Thus, in our study, exposure to physiological fluids for oocyte maturation and post-maturation does not improve in vitro embryo production in the horse.
    Language English
    Publishing date 2017-10
    Publishing country England
    Document type Journal Article
    ZDB-ID 1166294-3
    ISSN 1469-8730 ; 0967-1994
    ISSN (online) 1469-8730
    ISSN 0967-1994
    DOI 10.1017/S096719941700048X
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  9. Article ; Online: Effect of holding equine oocytes in meiosis inhibitor-free medium before in vitro maturation and of holding temperature on meiotic suppression and mitochondrial energy/redox potential.

    Martino, Nicola A / Dell'Aquila, Maria E / Filioli Uranio, Manuel / Rutigliano, Lucia / Nicassio, Michele / Lacalandra, Giovanni M / Hinrichs, Katrin

    Reproductive biology and endocrinology : RB&E

    2014  Volume 12, Page(s) 99

    Abstract: Background: Evaluation of mitochondrial function offers an alternative to evaluate embryo development for assessment of oocyte viability, but little information is available on the relationship between mitochondrial and chromatin status in equine ... ...

    Abstract Background: Evaluation of mitochondrial function offers an alternative to evaluate embryo development for assessment of oocyte viability, but little information is available on the relationship between mitochondrial and chromatin status in equine oocytes. We evaluated these parameters in immature equine oocytes either fixed immediately (IMM) or held overnight in an Earle's/Hank's' M199-based medium in the absence of meiotic inhibitors (EH treatment), and in mature oocytes. We hypothesized that EH holding may affect mitochondrial function and that holding temperature may affect the efficiency of meiotic suppression.
    Methods: Experiment 1 - Equine oocytes processed immediately or held in EH at uncontrolled temperature (22 to 27°C) were evaluated for initial chromatin configuration, in vitro maturation (IVM) rates and mitochondrial energy/redox potential. Experiment 2 - We then investigated the effect of holding temperature (25°C, 30°C, 38°C) on initial chromatin status of held oocytes, and subsequently repeated mitochondrial energy/redox assessment of oocytes held at 25°C vs. immediately-evaluated controls.
    Results: EH holding at uncontrolled temperature was associated with advancement of germinal vesicle (GV) chromatin condensation and with meiotic resumption, as well as a lower maturation rate after IVM. Holding did not have a significant effect on mitochondrial distribution within chromatin configurations. Independent of treatment, oocytes having condensed chromatin had a significantly higher proportion of perinuclear/pericortical mitochondrial distribution than did other GV configurations. Holding did not detrimentally affect oocyte energy/redox parameters in viable GV-stage oocytes. There were no significant differences in chromatin configuration between oocytes held at 25°C and controls, whereas holding at higher temperature was associated with meiosis resumption and loss of oocytes having the condensed chromatin GV configuration. Holding at 25°C was not associated with progression of mitochondrial distribution pattern and there were no significant differences in oocyte energy/redox parameters between these oocytes and controls.
    Conclusions: Mitochondrial distribution in equine GV-stage oocytes is correlated with chromatin configuration within the GV. Progression of chromatin configuration and mitochondrial status during holding are dependent on temperature. EH holding at 25°C maintains meiotic arrest, viability and mitochondrial potential of equine oocytes. This is the first report on the effects of EH treatment on oocyte mitochondrial energy/redox potential.
    MeSH term(s) Abattoirs ; Animals ; Cell Survival ; Chromatin Assembly and Disassembly ; Cold Temperature/adverse effects ; Culture Media ; Energy Metabolism ; Female ; Horses/physiology ; In Vitro Oocyte Maturation Techniques/veterinary ; Meiosis ; Microscopy, Confocal/veterinary ; Microscopy, Fluorescence/veterinary ; Mitochondria/metabolism ; Oocytes/cytology ; Oocytes/metabolism ; Oogenesis ; Oxidation-Reduction ; Reactive Oxygen Species/metabolism
    Chemical Substances Culture Media ; Reactive Oxygen Species
    Language English
    Publishing date 2014-10-11
    Publishing country England
    Document type Journal Article ; Research Support, Non-U.S. Gov't
    ISSN 1477-7827
    ISSN (online) 1477-7827
    DOI 10.1186/1477-7827-12-99
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  10. Article ; Online: Delta opioid receptor on equine sperm cells: subcellular localization and involvement in sperm motility analyzed by computer assisted sperm analyzer (CASA).

    Albrizio, Maria / Lacalandra, Giovanni M / Micera, Elisabetta / Guaricci, Antonio C / Nicassio, Michele / Zarrilli, Antonia

    Reproductive biology and endocrinology : RB&E

    2010  Volume 8, Page(s) 78

    Abstract: Background: Opioid receptors and endogenous opioid peptides act not only in the control of nociceptive pathways, indeed several reports demonstrate the effects of opiates on sperm cell motility and morphology suggesting the importance of these receptors ...

    Abstract Background: Opioid receptors and endogenous opioid peptides act not only in the control of nociceptive pathways, indeed several reports demonstrate the effects of opiates on sperm cell motility and morphology suggesting the importance of these receptors in the modulation of reproduction in mammals. In this study we investigated the expression of delta opioid receptors on equine spermatozoa by western blot/indirect immunofluorescence and its relationship with sperm cell physiology.
    Methods: We analyzed viability, motility, capacitation, acrosome reaction and mitochondrial activity in the presence of naltrindole and DPDPE by means of a computer assisted sperm analyzer and a fluorescent confocal microscope. The evaluation of viability, capacitation and acrosome reaction was carried out by the double CTC/Hoechst staining, whereas mitochondrial activity was assessed by means of MitoTracker Orange dye.
    Results: We showed that in equine sperm cells, delta opioid receptor is expressed as a doublet of 65 and 50 kDa molecular mass and is localized in the mid piece of tail; we also demonstrated that naltrindole, a delta opioid receptor antagonist, could be utilized in modulating several physiological parameters of the equine spermatozoon in a dose-dependent way. We also found that low concentrations of the antagonist increase sperm motility whereas high concentrations show the opposite effect. Moreover low concentrations hamper capacitation, acrosome reaction and viability even if the percentage of cells with active mitochondria seems to be increased; the opposite effect is exerted at high concentrations. We have also observed that the delta opioid receptor agonist DPDPE is scarcely involved in affecting the same parameters at the employed concentrations.
    Conclusions: The results described in this paper add new important details in the comprehension of the mammalian sperm physiology and suggest new insights for improving reproduction and for optimizing equine breeding.
    MeSH term(s) Animals ; Cell Survival/drug effects ; Enkephalin, D-Penicillamine (2,5)-/pharmacology ; Horses/metabolism ; Image Processing, Computer-Assisted/methods ; Male ; Naltrexone/analogs & derivatives ; Naltrexone/pharmacology ; Receptors, Opioid, delta/agonists ; Receptors, Opioid, delta/antagonists & inhibitors ; Receptors, Opioid, delta/metabolism ; Semen Analysis/instrumentation ; Semen Analysis/methods ; Sperm Motility/drug effects ; Sperm Motility/physiology ; Spermatozoa/drug effects ; Spermatozoa/metabolism ; Subcellular Fractions/metabolism ; Tissue Distribution/drug effects
    Chemical Substances Receptors, Opioid, delta ; Naltrexone (5S6W795CQM) ; Enkephalin, D-Penicillamine (2,5)- (88373-73-3) ; naltrindole (G167Z38QA4)
    Language English
    Publishing date 2010-06-25
    Publishing country England
    Document type Journal Article
    ZDB-ID 2119215-7
    ISSN 1477-7827 ; 1477-7827
    ISSN (online) 1477-7827
    ISSN 1477-7827
    DOI 10.1186/1477-7827-8-78
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