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2015 | 18 | 3 |

Tytuł artykułu

Apoptotic - like changes of boar spermatozoa in freezing media supplemented with different antioxidants

Treść / Zawartość

Warianty tytułu

Języki publikacji

EN

Abstrakty

EN
This study evaluated the effect of supplementing the freezing extender with exogenous antioxidants on apoptotic-like changes in post-thaw boar spermatozoa. A total of 36 ejaculates were resuspended in standard lactose-egg yolk-glycerol extender supplemented with antioxidant to final concentrations of 0 (as control), 2.5mM GSH (group I), 5.0 mM GSH (group II), 150 IU/mL SOD (group III), 300 IU/mL SOD (group IV), 200 IU/mL CAT (group V), 400 IU/mL CAT (group VI), 150 IU/mL SOD+200 IU/mL CAT (group VII), 300 IU/mL SOD+400 IU/mL CAT (group VIII). Sperm motility and apoptotic-like changes were determined before and after freeze-thawing. The various markers of apoptotic-like changes were measured: plasma membrane permeability by YO-PRO-1/PI assay, phosphatidylserine (PS) translocation across the plasma membrane using fluorescein-labeled Annexin-V, mitochondrial transmembrane potential detected by JC-1, and DNA fragmentation evaluated by TUNEL assay. The highest percentage of progressive motile sperm was noticed in group II (PM% 64.2±15.4) compared with control (PM% 36.8±5.5). The supplementation of 400 IU/mL CAT (group VI) revealed significant (P<0.01) reduction of apoptotic-like changes (YO-PRO-1⁺/PI⁻: 13.1±7.5%, AnV⁺/PI⁻: 9.9±4.1%) in frozen-thawed spermatozoa compared with extendersupplemented with200 IU/mL CAT (groupV). Irrespectiveof the concentrationused, SOD and CAT in combination (group VII and group VIII) significantly (P<0.01) improved post-thaw sperm survival compared with the control. Evaluation by TUNEL assay revealed that cryopreservation and thawing did not induce DNA fragmentation in boar spermatozoa.

Słowa kluczowe

Wydawca

-

Rocznik

Tom

18

Numer

3

Opis fizyczny

p.473-480,ref.

Twórcy

autor
  • Department of Biotechnology of Animal Reproduction, National Research Institute of Animal Production, Krakowska 1, 32-083 Balice/Krakow, Poland
autor
  • Department of Biotechnology of Animal Reproduction, National Research Institute of Animal Production, Krakowska 1, 32-083 Balice/Krakow, Poland

Bibliografia

  • Aitken RJ (1995) Free radicals, lipid peroxidation and sperm function. Reprod Fertil Dev 7: 659-668.
  • Alvarez JG, Storey BT (1992) Evidence for increased lipid peroxidative damage and loss of superoxide dismutase activity as a mode of sublethal cryodamage to human sperm during cryopreservation. J Androl 13: 232-241.
  • Anzar M, He L, Buhr MM, Kroetsch TG, Pauls KP (2002) Sperm apoptosis in fresh and cryopreserved bull semen detected by flow cytometry and its relationship with fertility. Biol Reprod 66: 354-360.
  • Bryła M, Trzcińska M, Wieczorek J, Słomski R, Smorąg Z (2010) Effect of semen quality in transgenic boars on the developmental competence of preimplantation embryos. Anim Reprod Sci 18: 77-82.
  • Duru NK, Morshedi M, Schuffner A, Oehninger S (2001) Cryopreservation-thawing of fractionated human spermatozoa is associated with membrane phosphatidylserine externalization and not DNA fragmentation. J Androl 22: 646-651.
  • Fraser L, Strzeżek J (2007) Is there a relationship between the chromatin status and DNA fragmentation of boar spermatozoa following freezing-thawing? Theriogenology 68: 248-257.
  • Fraser L, Strzeżek J, Kordan W (2011) Effect of freezing on sperm nuclear DNA. Reprod Domest Anim 46 (Suppl 2): 14-17.
  • Gadea J, Selles E, Marco MA, Coy P, Matas C, Romar R, Ruiz S (2004) Decrease in glutathione content in boar sperm after cryopreservation. Effect of the addition of reduced glutathione to the freezing and thawing extenders. Theriogenology 62: 690-701.
  • Gadea J, Garcta-Vazquez F, Maths C, Gardón JC, Chnovas S, Gumbao D (2005) Cooling and freezing of boar spermatozoa: supplementation of the freezing media with reduced glutathione preserves sperm function. J Androl 26: 396-404.
  • Kadirvel G, Periasamy S, Kumar S (2012) Effect of cryopreservation on apoptotic-like events and its relationship with cryocapacitation of buffalo (Bubalus bubalis) sperm. Reprod Domest Anim 47: 143-150.
  • Katari S, Turan N, Bibikova M, Erinle O, Chalian R, Foster M , Gaughan JP, Coutifaris C, Sapienza C (2009) DNA methylation and gene expression differences in children conceived in vitro or in vivo. Hum Mol Genet 18: 3769-3778.
  • Kowalowka M, Wysocki P, Fraser L, Strzeżek J (2008) Extracellular superoxide dismutase of boar seminal plasma. Reprod Domest Anim 43: 490-496.
  • Koziorowska-Gilun M, Koziorowski M, Fraser L, Strzeżek J (2011) Antioxidant defence system of boar cauda epididymidal spermatozoa and reproductive tract fluids. Reprod Domest Anim 46: 527-533.
  • Martin G, Sabido O, Durand P, Levy R (2004) Cryopreservation induces an apoptosis-like mechanism in bull sperm. Biol Reprod 71: 28-37.
  • Ortega-Ferrusola C, Sotillo-Galhn Y, Varela-Fernhndez E, Gallardo-Bolanos JM, Muriel A, Gonzhlez-Fernhndez L, Tapia JA, Pena FJ (2008) Detection of „apoptosis-like” changes during the cryopreservation process in equine sperm. J Androl 29: 213-221.
  • Pena FJ, Johannisson A, Wallgren M, Rodriguez-Martinez H (2003) Assessment of fresh and frozen-thawed boar semen using an Annexin-V assay: a new method of evaluating sperm membrane integrity. Theriogenology 60: 677-689.
  • Roca J, Rodriguez MJ, Gil MA, Carvajal G, Garcia EM, Cuello C, Vazquez JM, Martinez EA (2005) Survival and in vitro fertility of boar spermatozoa frozen in the presence of superoxide dismutase and/or catalase. J Androl 26: 15-24.
  • Rodrtguez-Marttnez H (2003) Laboratory semen assessment and prediction of fertility: still utopia? Reprod Domest Anim 38: 312-318.
  • Silva PF, Gadella BM (2006) Detection of damage in mammalian sperm cell. Theriogenology 65: 958-978.
  • Trzcińska M, Bryła M, Gogol P, Cegła M (2013) Kriokonserwacja nasienia knura. ISBN 978-83-7607-296-8. Zespół Wydawnictw i Poligrafii IZ PIB, pp 1-12.
  • Trzcińska M, Bryła M, Gajda B, Gogol P (2015) Fertility of boar semen cryopreserved in extender supplemented with butylated hydroxytoluene. Theriogenology 83: 307-313.
  • Yeste M, Estrada E, Pinart E, Bonet S, Miró J, Rodriquez-Gil JE (2014) The improving effect of reduced glutathione on boar sperm cryotolerance is related with the intrinsic ejaculate freezability. Cryobiology 68: 251-261.

Typ dokumentu

Bibliografia

Identyfikatory

Identyfikator YADDA

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