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Çözüm sonu teke spermasına ATP ilavesinin sperma parametreleri üzerine etkisi

Yıl 2025, Cilt: 14 Sayı: 2, 46 - 51, 25.12.2025
https://doi.org/10.53913/aduveterinary.1809432

Öz

Çözüm sonrası teke spermatozoonlarına farklı doz seviyelerinde ATP takviyesinin motilite ve diğer sperm parametreleri üzerindeki etkisi 4 saatlik inkübasyon süresince değerlendirildi. Ejekülat, yumurta sarısı-glikoz-gliserol-tris-sitrik asit sulandırıcısı kullanılarak 0.25 ml payetlerde donduruldu. Su banyosunda 30 saniye süreyle 37°C'de payetler çözüldü ve sperm hücreleri, sulandırıcı içeriklerinin uzaklaştırılmasının ardından 0 (kontrol), 2, 4 veya 8 mM ATP eklenmeden önce tris-sitrik asit-glikoz çözeltisi içerisinde yeniden seyreltildi. Spermatolojik parametrelerin büyük çoğunluğu ATP takviyesinden (P<0.05) olumlu biçimde etkilenmiştir. ATP takviyesi (2-4 mM), orta hızda progresif sperm alt popülasyonunun yüzdesini artırarak toplam progresif hareketliliği yükseltti (P<0.05). Ancak ATP takviyesi ALH, VAP, VCL ve VSL değerlerini artırmadı. Buna karşılık, ATP' nin aşırı doz seviyeleri (8 mM), ALH, VAP, VCL ve VSL dahil olmak üzere hız parametrelerinin çoğunu (P<0.001) baskıladı. Ölü-canlı, akrozom reaksiyonu oranı ve mitokondriyal aktivite doz seviyelerine bakılmaksızın ATP takviyesinden etkilenmedi (P>0.05). Sonuç olarak, çözüm sonrası teke spermatozoasına ATP takviyesi (2-4 mM), ALH, VCL, VAP ve VSL dahil olmak üzere kinematik sperm parametrelerinin çoğu üzerinde belirgin bir etki olmaksızın inkübasyon periyodu boyunca progresif motiliteyi geliştirdi (P<0.05). ATP konsantrasyonunun 8 mM seviyesine çıkarılması motilite parametrelerini önemli ölçüde düşürdü. (P<0.01). ATP takviyesinin, teke spermatozoasında kriyoprezervasyondan kaynaklanan azalmış motilite problemini en azından kısmen düzeltebileceği sonucuna varıldı.

Etik Beyan

Tüm deneysel prosedürler, Aydın Adnan Menderes Üniversitesi Hayvan Deneyleri Etik Kurulu tarafından incelenmiş ve 64583101/2023/147 protokol numarası ile onaylanmıştır.

Teşekkür

Tekelerin barındırılması konusunda sağladıkları destek için Aydın Adnan Menderes Üniversitesi Veteriner Fakültesine ve laboratuvar çalışmalarının yürütüldüğü Dölerme ve Suni Tohumlama Anabilim Dalına teşekkür ederiz.

Kaynakça

  • Agarwal, A., & Prabakaran, S.A. (2005). Mechanism, measurement, and prevention of oxidative stress in male reproductive physiology. Indian Journal of Experimental Biology, 43, 963–974.
  • Agarwal, A., Virk, G., Ong, C., & Du Plessis, S. S. (2014). Effect of oxidative stress on male reproduction. World Journal of Men’s Health, 32(1), Uçan et al ATP Addition to Goat Semen 51 1–17. https://doi.org/10.5534/wjmh.2014.32.1.1
  • Aitken, R.J., Sutton, M., Warner, P., & Richardson, D.W. (1985). Relationship between the movement characteristics of human spermatozoa and their ability to penetrate cervical mucus and zona-free hamster oocytes. Journal of Reproduction and Fertility, 73, 441–449. https://doi.org/10.1530/jrf.0.0730441
  • Berg, J.M., Tymoczko, J.L., & Stryer, L. (2002). Biochemistry (5th ed.). New York, NY: W.H. Freeman. Section 16.1, Glycolysis is an energyconversion pathway in many organisms. 648-668
  • Bonora, M., Patergnani, S., Rimessi, A., De Marchi, E., Suski, J.M., Bononi, A.,...& Pinton, P. (2012). ATP synthesis and storage. Purinergic Signalling, 8, 343–357. https://doi.org/10.1007/s11302-012-9305-8
  • De Lamirande, E., & Gagnon, C. (1992). Reactive oxygen species and human spermatozoa. II. Depletion of adenosine triphosphate plays an important role in the inhibition of sperm motility. Journal of Andrology, 13, 379–386.
  • Du Plessis, S.S., Agarwal, A., Mohanty, G., & van der Linde, M. (2015). Oxidative phosphorylation versus glycolysis: What fuel do spermatozoa use? Asian Journal of Andrology, 17(2), 230–235. https://doi.org/10.4103/1008-682X.135123
  • Evans, G., & Maxwell, W.M. C. (1987). Salamon’s artificial insemination of sheep and goats. London: Butterworths. Chapter 14, pp. 122–144.
  • Farrell, P.B., Foote, R.H., McArdle, M.M., Trouern-Trend, V.L., & Tardif, A.L. (1996). Media and dilution procedures tested to minimize handling effects on human, rabbit, and bull sperm for computerassisted sperm analysis (CASA). Journal of Andrology, 17, 293–300.
  • Flores, E., Fernández-Novell, J.M., Peña, A., Rigau, T., & RodríguezGil, J.E. (2010). Cryopreservation-induced alterations in boar spermatozoa mitochondrial function are related to changes in the expression and location of midpiece mitofusin-2 and actin network. Theriogenology, 74, 354–363. https://doi.org/10.1016/j.theriogenology.2010.02.018.
  • Florman, H.M., Arnoult, C., Kazam, I.G., Li, C., & O’Toole, C.M. (1998). A perspective on the control of mammalian fertilization by egg-activated ion channels in sperm: A tale of two channels. Biology of Reproduction, 59, 12–16. https://doi.org/10.1095/biolreprod59.1.12.
  • Foresta, C., Rossato, M., & Di Virgilio, F. (1992). Extracellular ATP is a trigger for the acrosome reaction in human spermatozoa. Journal of Biological Chemistry, 267, 19443–19447. https://doi.org/10.1016/S0021-9258(18)41795-4
  • Hammerstedt, R.H. (1981). Monitoring the metabolic rate of germ cells and sperm. In K. McKern (Ed.), Reproductive processes and contraception (pp. 353–372). New York, NY: Plenum Press.
  • Holt, W.V., Head, M.F., & North, R.D. (1992). Freezeinduced membrane damage in ram spermatozoa is manifested after thawing: Observations with experimental cryomicroscopy. Biology of Reproduction, 46, 1086–1094. https://doi.org/10.1095/biolreprod46.6.1086
  • Karuhn, R.F. (1977). Method of predetermining time of ovulation in women and in animals to control conception (U.S. Patent No. 4,036,212).
  • Kim, S., Hooper, S., Ağca, C., & Ağca, Y. (2016). Post-thaw supplementation enhances cryoprotective effect of iodixanol in rat spermatozoa. Reproductive Biology and Endocrinology, 14, 5. https://doi.org/10.1186/s12958-016-0141-5
  • Lindemann, C.B., Fisher, M., & Lipton, M. (1982). A comparative study of the effects of freezing and frozen storage on intact and demembranated bull spermatozoa. Cryobiology, 19, 20–28.
  • Long, J.A. (2006). Avian semen cryopreservation: What are the biological challenges? Poultry Science, 85, 232–236. https://doi.org/10.1093/ps/85.2.232
  • Luria, A., Rubinstein, S., Lax, Y., & Breitbart, H. (2002). Extracellular adenosine triphosphate stimulates acrosomal exocytosis in bovine spermatozoa via P2 purinoreceptor. Biology of Reproduction, 66, 429–437.
  • O’Connell, M., McClure, N., & Lewis, S.E. (2002). The effects of cryopreservation on sperm morphology, motility and mitochondrial function. Human Reproduction, 17, 704–709. https://doi.org/10.1093/humrep/17.3.704
  • Perumal, P., Selvaraju, S., Selvakumar, S., Barik, A.K., Mohanty, D.N., Nas, S.,...& Mishra, P.C. (2011). Effect of prefreeze addition of cysteine hydrochloride and reduced glutathione in semen of crossbred Jersey bulls on sperm parameters and conception rates. Reproduction in Domestic Animals, 46, 636–641.
  • Rajender, S., Rahul, P., & Mahdi, A.A. (2010). Mitochondria, spermatogenesis and male infertility. Mitochondrion, 10, 419–428. https://doi.org/10.1016/j.mito.2010.05.015
  • Rodriguez-Miranda, E., Buffone, M.G., Edwards, S.E., Ord, T.S., Lin, K., Sammel, M.D.,...& Williams, C.J. (2008). Extracellular adenosine 5′-triphosphate alters motility and improves the fertilizing capability of mouse sperm. Biology of Reproduction, 79, 164–171.
  • Rossato, M., La Sala, G.B., Balasini, M., Taricco, F., Galeazzi, C., Ferlin, A., & Foresta, C. (1999). Sperm treatment with extracellular ATP increases fertilization rates in in-vitro fertilization for male factor infertility. Human Reproduction, 14, 694–697. https://doi.org/10.1093/humrep/14.3.694
  • Ruiz-Pesini, E., Díez-Sánchez, C., López-Pérez, M.J., & Enríquez, J.A. (2007). The role of the mitochondrion in sperm function: Is there a place for oxidative phosphorylation or is this a purely glycolytic process? Current Topics in Developmental Biology, 77, 3–19. https://doi.org/10.1016/S0070-2153(06)77001-6
  • Saleh, R.A., Agarwal, A., Sharma, R.K., Nelson, D.R., & Thomas, A.J. (2002). Effect of cigarette smoking on levels of seminal oxidative stress in infertile men: A prospective study. Fertility and Sterility, 78, 491–499.
  • Schober, D., Aurich, C., Nohl, H., & Gille, L. (2007). Influence of cryopreservation on mitochondrial functions in equine spermatozoa. Theriogenology, 68, 745–754. https://doi.org/10.1016/j.theriogenology.2007.06.004
  • Shariatmadari, R., Sipilä, P., Vierula, M., Törnquist, K., Huhtaniemi, I., & Poutanen, M. (2003). Adenosine triphosphate induces Ca²⁺ signal in epithelial cells of the mouse caput epididymis through activation of P2X and P2Y purinergic receptors. Biology of Reproduction, 68, 1185–1192. https://doi.org/10.1095/biolreprod.102.007419
  • Storey, B.T. (2008). Mammalian sperm metabolism: Oxygen and sugar, friend and foe. The International Journal of Developmental Biology,52,427–437. https://doi.org/10.1387/ijdb.072522bs
  • Thuwanut, P., Arya, N., Comizzoli, P., & Chatdarong, K. (2015). Effect of extracellular adenosine 5′-triphosphate on cryopreserved epididymal cat sperm intracellular ATP concentration, sperm quality, and in vitro fertilizing ability. Theriogenology, 84, 702–709.
  • Thuwanut, P., Chatdarong, K., Johannisson, A., Bergqvist, A.S., Söderquist, L., & Axnér, E. (2010). Cryopreservation of epididymal cat spermatozoa: Effects of in vitro antioxidative enzyme supplementation and lipid peroxidation induction. Theriogenology, 73, 1076–1087. https://doi.org/10.1016/j.theriogenology.2010.01.007
  • Turner, R.M. (2003). Tales from the tail: What do we really know about sperm motility? Journal of Andrology, 24, 790–803. https://doi.org/10.1002/j.1939-4640.2003.tb03123.x
  • Yamashiro, H., Toyomizu, M., Kikusato, M., Toyama, N., Sugimura, S., Hoshino, Y.,...& Sato, E. (2010). Lactate and adenosine triphosphate in the extender enhance the cryosurvival of rat epididymal sperm. Journal of the American Association for Laboratory Animal Science, 49, 160–166.

Effect of ATP Addition to Goat Semen Post Thaw on Semen Parameters

Yıl 2025, Cilt: 14 Sayı: 2, 46 - 51, 25.12.2025
https://doi.org/10.53913/aduveterinary.1809432

Öz

In post-thaw goat spermatozoa the impact of ATP supplementation at different dose levels on motility and the other sperm parameters was evaluated during 4 hours of incubation. The ejaculate was frozen in 0.25ml straws by using egg yolk-glucose-glycerol-tris-citric acid extender. Thawing was achieved at 37°C for 30 second in a water bath and sperm cells were rediluted, following the removal of the extender, in tris-citric acid-glucose solution before adding 0 (control), 2, 4 or 8 mM of ATP. Eight of 10 parameters that were examined by the computer assisted sperm analyzer has been influenced by ATP supplementation (P<0.05) at dose dependent manner. ATP supplementation (2-4mM) increased (P<0.05) progressive motility by enhancing the percentage of medium progressive sperm subpopulation (P<0.05). However, ATP supplementation did not increase ALH, VAP, VCL and VSL values. In contrast, excessive dose levels of ATP (8mM) depressed most of the velocity parameters (P<0.001) including ALH, VAP, VCL and VSL. The sperm parameters other than motility, live and acrosome reacted sperm rate and mitochondrial activity were not influenced (P>0.05) by ATP supplementation irrespective to the dose levels.
As a result, ATP supplementation (2-4mM) to post-thaw goat spermatozoa improved progressive motility during the incubation (P<0.05) without any prominent influence on most of the kinematic sperm parameters including ALH, VCL, VAP and VSL. Further increase the ATP concentrations to 8mM suppressed motility parameters significantly (P<0.01). It was concluded that ATP supplementation might restore reduced motility problem, at least partly, in goat spermatozoa resulting from cryopreservation.

Etik Beyan

All study procedures were reviewed and approved by the Animal Research Ethics Committee of the Aydin Adnan Menderes University, under protocol number 64583101/2023/147.

Teşekkür

We would like to thank Aydın Adnan Menderes University, Faculty of Veterinary Medicine, for providing assistance with the housing of the bucks, and the Department of Reproduction and Artificial Insemination, where the laboratory studies were conducted.

Kaynakça

  • Agarwal, A., & Prabakaran, S.A. (2005). Mechanism, measurement, and prevention of oxidative stress in male reproductive physiology. Indian Journal of Experimental Biology, 43, 963–974.
  • Agarwal, A., Virk, G., Ong, C., & Du Plessis, S. S. (2014). Effect of oxidative stress on male reproduction. World Journal of Men’s Health, 32(1), Uçan et al ATP Addition to Goat Semen 51 1–17. https://doi.org/10.5534/wjmh.2014.32.1.1
  • Aitken, R.J., Sutton, M., Warner, P., & Richardson, D.W. (1985). Relationship between the movement characteristics of human spermatozoa and their ability to penetrate cervical mucus and zona-free hamster oocytes. Journal of Reproduction and Fertility, 73, 441–449. https://doi.org/10.1530/jrf.0.0730441
  • Berg, J.M., Tymoczko, J.L., & Stryer, L. (2002). Biochemistry (5th ed.). New York, NY: W.H. Freeman. Section 16.1, Glycolysis is an energyconversion pathway in many organisms. 648-668
  • Bonora, M., Patergnani, S., Rimessi, A., De Marchi, E., Suski, J.M., Bononi, A.,...& Pinton, P. (2012). ATP synthesis and storage. Purinergic Signalling, 8, 343–357. https://doi.org/10.1007/s11302-012-9305-8
  • De Lamirande, E., & Gagnon, C. (1992). Reactive oxygen species and human spermatozoa. II. Depletion of adenosine triphosphate plays an important role in the inhibition of sperm motility. Journal of Andrology, 13, 379–386.
  • Du Plessis, S.S., Agarwal, A., Mohanty, G., & van der Linde, M. (2015). Oxidative phosphorylation versus glycolysis: What fuel do spermatozoa use? Asian Journal of Andrology, 17(2), 230–235. https://doi.org/10.4103/1008-682X.135123
  • Evans, G., & Maxwell, W.M. C. (1987). Salamon’s artificial insemination of sheep and goats. London: Butterworths. Chapter 14, pp. 122–144.
  • Farrell, P.B., Foote, R.H., McArdle, M.M., Trouern-Trend, V.L., & Tardif, A.L. (1996). Media and dilution procedures tested to minimize handling effects on human, rabbit, and bull sperm for computerassisted sperm analysis (CASA). Journal of Andrology, 17, 293–300.
  • Flores, E., Fernández-Novell, J.M., Peña, A., Rigau, T., & RodríguezGil, J.E. (2010). Cryopreservation-induced alterations in boar spermatozoa mitochondrial function are related to changes in the expression and location of midpiece mitofusin-2 and actin network. Theriogenology, 74, 354–363. https://doi.org/10.1016/j.theriogenology.2010.02.018.
  • Florman, H.M., Arnoult, C., Kazam, I.G., Li, C., & O’Toole, C.M. (1998). A perspective on the control of mammalian fertilization by egg-activated ion channels in sperm: A tale of two channels. Biology of Reproduction, 59, 12–16. https://doi.org/10.1095/biolreprod59.1.12.
  • Foresta, C., Rossato, M., & Di Virgilio, F. (1992). Extracellular ATP is a trigger for the acrosome reaction in human spermatozoa. Journal of Biological Chemistry, 267, 19443–19447. https://doi.org/10.1016/S0021-9258(18)41795-4
  • Hammerstedt, R.H. (1981). Monitoring the metabolic rate of germ cells and sperm. In K. McKern (Ed.), Reproductive processes and contraception (pp. 353–372). New York, NY: Plenum Press.
  • Holt, W.V., Head, M.F., & North, R.D. (1992). Freezeinduced membrane damage in ram spermatozoa is manifested after thawing: Observations with experimental cryomicroscopy. Biology of Reproduction, 46, 1086–1094. https://doi.org/10.1095/biolreprod46.6.1086
  • Karuhn, R.F. (1977). Method of predetermining time of ovulation in women and in animals to control conception (U.S. Patent No. 4,036,212).
  • Kim, S., Hooper, S., Ağca, C., & Ağca, Y. (2016). Post-thaw supplementation enhances cryoprotective effect of iodixanol in rat spermatozoa. Reproductive Biology and Endocrinology, 14, 5. https://doi.org/10.1186/s12958-016-0141-5
  • Lindemann, C.B., Fisher, M., & Lipton, M. (1982). A comparative study of the effects of freezing and frozen storage on intact and demembranated bull spermatozoa. Cryobiology, 19, 20–28.
  • Long, J.A. (2006). Avian semen cryopreservation: What are the biological challenges? Poultry Science, 85, 232–236. https://doi.org/10.1093/ps/85.2.232
  • Luria, A., Rubinstein, S., Lax, Y., & Breitbart, H. (2002). Extracellular adenosine triphosphate stimulates acrosomal exocytosis in bovine spermatozoa via P2 purinoreceptor. Biology of Reproduction, 66, 429–437.
  • O’Connell, M., McClure, N., & Lewis, S.E. (2002). The effects of cryopreservation on sperm morphology, motility and mitochondrial function. Human Reproduction, 17, 704–709. https://doi.org/10.1093/humrep/17.3.704
  • Perumal, P., Selvaraju, S., Selvakumar, S., Barik, A.K., Mohanty, D.N., Nas, S.,...& Mishra, P.C. (2011). Effect of prefreeze addition of cysteine hydrochloride and reduced glutathione in semen of crossbred Jersey bulls on sperm parameters and conception rates. Reproduction in Domestic Animals, 46, 636–641.
  • Rajender, S., Rahul, P., & Mahdi, A.A. (2010). Mitochondria, spermatogenesis and male infertility. Mitochondrion, 10, 419–428. https://doi.org/10.1016/j.mito.2010.05.015
  • Rodriguez-Miranda, E., Buffone, M.G., Edwards, S.E., Ord, T.S., Lin, K., Sammel, M.D.,...& Williams, C.J. (2008). Extracellular adenosine 5′-triphosphate alters motility and improves the fertilizing capability of mouse sperm. Biology of Reproduction, 79, 164–171.
  • Rossato, M., La Sala, G.B., Balasini, M., Taricco, F., Galeazzi, C., Ferlin, A., & Foresta, C. (1999). Sperm treatment with extracellular ATP increases fertilization rates in in-vitro fertilization for male factor infertility. Human Reproduction, 14, 694–697. https://doi.org/10.1093/humrep/14.3.694
  • Ruiz-Pesini, E., Díez-Sánchez, C., López-Pérez, M.J., & Enríquez, J.A. (2007). The role of the mitochondrion in sperm function: Is there a place for oxidative phosphorylation or is this a purely glycolytic process? Current Topics in Developmental Biology, 77, 3–19. https://doi.org/10.1016/S0070-2153(06)77001-6
  • Saleh, R.A., Agarwal, A., Sharma, R.K., Nelson, D.R., & Thomas, A.J. (2002). Effect of cigarette smoking on levels of seminal oxidative stress in infertile men: A prospective study. Fertility and Sterility, 78, 491–499.
  • Schober, D., Aurich, C., Nohl, H., & Gille, L. (2007). Influence of cryopreservation on mitochondrial functions in equine spermatozoa. Theriogenology, 68, 745–754. https://doi.org/10.1016/j.theriogenology.2007.06.004
  • Shariatmadari, R., Sipilä, P., Vierula, M., Törnquist, K., Huhtaniemi, I., & Poutanen, M. (2003). Adenosine triphosphate induces Ca²⁺ signal in epithelial cells of the mouse caput epididymis through activation of P2X and P2Y purinergic receptors. Biology of Reproduction, 68, 1185–1192. https://doi.org/10.1095/biolreprod.102.007419
  • Storey, B.T. (2008). Mammalian sperm metabolism: Oxygen and sugar, friend and foe. The International Journal of Developmental Biology,52,427–437. https://doi.org/10.1387/ijdb.072522bs
  • Thuwanut, P., Arya, N., Comizzoli, P., & Chatdarong, K. (2015). Effect of extracellular adenosine 5′-triphosphate on cryopreserved epididymal cat sperm intracellular ATP concentration, sperm quality, and in vitro fertilizing ability. Theriogenology, 84, 702–709.
  • Thuwanut, P., Chatdarong, K., Johannisson, A., Bergqvist, A.S., Söderquist, L., & Axnér, E. (2010). Cryopreservation of epididymal cat spermatozoa: Effects of in vitro antioxidative enzyme supplementation and lipid peroxidation induction. Theriogenology, 73, 1076–1087. https://doi.org/10.1016/j.theriogenology.2010.01.007
  • Turner, R.M. (2003). Tales from the tail: What do we really know about sperm motility? Journal of Andrology, 24, 790–803. https://doi.org/10.1002/j.1939-4640.2003.tb03123.x
  • Yamashiro, H., Toyomizu, M., Kikusato, M., Toyama, N., Sugimura, S., Hoshino, Y.,...& Sato, E. (2010). Lactate and adenosine triphosphate in the extender enhance the cryosurvival of rat epididymal sperm. Journal of the American Association for Laboratory Animal Science, 49, 160–166.
Toplam 33 adet kaynakça vardır.

Ayrıntılar

Birincil Dil İngilizce
Konular Hayvan Üreme ve Islahı
Bölüm Araştırma Makalesi
Yazarlar

Uğur Uçan 0000-0001-8325-138X

Zahid Naseer 0000-0001-6335-9633

Melih Aksoy 0000-0001-9573-4493

Gönderilme Tarihi 23 Ekim 2025
Kabul Tarihi 28 Kasım 2025
Yayımlanma Tarihi 25 Aralık 2025
Yayımlandığı Sayı Yıl 2025 Cilt: 14 Sayı: 2

Kaynak Göster

APA Uçan, U., Naseer, Z., & Aksoy, M. (2025). Effect of ATP Addition to Goat Semen Post Thaw on Semen Parameters. Animal Health Production and Hygiene, 14(2), 46-51. https://doi.org/10.53913/aduveterinary.1809432