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Flow Cytometric Examination of The Interaction Between Drone Sperm (Apis Mellifera Anatoliaca) and Insulin in Soybean Lecithin-Based Cryopreservation Extenders

Yıl 2024, Cilt: 43 Sayı: 1, 46 - 54, 02.09.2024
https://doi.org/10.30782/jrvm.1420188

Öz

This study was conducted to evaluate the effects of different doses of insulin (5, 10, and 15 IU) added to soybean lecithin-based drone sperm freezing extenders on sperm parameters through flow cytometry or microscopic examinations. Samples obtained from sexually mature bees were pooled to eliminate individual variation. The freezing process was organized with three insulin supplemented doses and one insulin-free control group, and the method was implemented as planned. Thawed samples were evaluated for motility, plasma membrane (hypo-osmotic swelling test), and DNA integrity (TUNEL assay) using a phase-contrast microscope with fluorescent attachment. Additionally, acrosomal integrity (PNA-FITC), nitric oxide levels (DAF-2/DA), and mitochondrial membrane potential (JC-1) were investigated using a flow cytometer. As expected, all sperm parameters were affected by the freezing process inherently, however, the insulin-supplemented experimental groups did not demonstrate a statistically significant positive difference compared to the control group. In light of these results, it was determined that the use of insulin supplementation, in preferred doses, in soybean lecithin-based drone sperm cryopreservation extenders did not have a positive effect on achieving higher productive honeybee colonies and elevating the country's yield in bee products through auxiliary biotechnological methods as desired.

Proje Numarası

Bursa Uludağ Üniversitesi Bilimsel Araştırma Projeleri Birimi (BAP) - TGA – 2022 - 718

Kaynakça

  • Henry LC, Álvaro D rebolledo, Karen C medina, Jorge V rodríguez, Francisco E valente. Effect of egg yolk on sperm cryopreservation of drone (Apis mellifera). Abanico Vet. 2019;9(December):1-11. doi:10.21929/ abavet2019.921
  • Stankus T. A review and bibliography of the literature of honey bee colony collapse disorder: A poorly understood epidemic that Clearly Threatens the successful pollination of billions of dollars of crops in America. J Agric Food Inf. 2008;9(2):115-143. doi:10.1080/10496500802173939
  • Gül A, Şahinler N, Onal AG, Hopkins BK, Sheppard WS. Effects of diluents and plasma on honey bee (Apis mellifera L.) drone frozen-thawed semen fertility. Theriogenology. 2017;101:109-113. doi:10.1016/j. theriogenology.2017.06.020
  • Woyke J, Jasinski Z. Influence of Age of Drones on the Results of Instrumental Insemination of Honeybee Queens. Apidologie. 1978;9(3):203-212. doi:10.1051/ apido:19780304
  • Cobey SW. Comparison studies of instrumentally inseminated and naturally mated honey bee queens and factors affecting their performance. Apidologie. 2007;38(4):390-410. doi:10.1051/apido:2007029
  • Alçay S, Üstüner B, Çakmak İ, Çakmak S, Nur Z. Effects of Various Cryoprotective Agents on Post-Thaw Drone Semen Quality. Kafkas Univ Vet Fak Derg. 2015;21(1):31-35. doi:10.9775/kvfd.2014.11515
  • Almadaly EA, Tawfik FS, El-Kon II, Heleil BA, Fattouh ESM. Effect of different cryoprotectants on the post-thaw sperm characteristics and in vivo fertility of buffalo (Bubalus bubalis) bull semen. Slov Vet Res. 2019;56(February):541-551. doi:10.26873/SVR-792-2019
  • Tonieto RA, Goularte KL, Gastal GDA, Schiavon RS, Deschamps JC, Lucia T. Cryoprotectant effect of trehalose and low-density lipoprotein in extenders for frozen ram semen. Small Rumin Res. 2010;93(2-3):206- 209. doi:10.1016/j.smallrumres.2010.05.003
  • Abdelkader F Ben, Kairo G, Bonnet M, Barbouche N, Belzunces LP, Brunet JL. Effects of clothianidin on antioxidant enzyme activities and malondialdehyde level in honey bee drone semen. J Apic Res. 2019;58(5):740- 745. doi:10.1080/00218839.2019.1655182
  • Kulaksiz R, Daskin AD. In vitro evaluation of Saanen buck semen frozen in different extenders supplemented with various antioxidants. Ankara Üniv Vet Fak Derg. 2010;57:151-156. doi:10.1501/Vetfak_0000002369
  • Borsuk G, Kozłowska M, Anusiewicz M, Paleolog J. Nosema ceranae changes semen characteristics and damages sperm DNA in honeybee drones. Invertebr Surviv J. 2018;15:197-202.
  • Ciereszko A, Wilde J, Dietrich GJ, et al. Sperm parameters of honeybee drones exposed to imidacloprid. Apidologie. 2017;48(2):211-222. doi:10.1007/s13592- 016-0466-2
  • Toker MB, Alcay S, Gokce E, Ustuner B. Cryopreservation of ram semen with antioxidant supplemented soybean lecithin-based extenders and impacts on incubation resilience. Cryobiology. 2016;72(3):205-209. doi:10.1016/j.cryobiol.2016.05.001
  • Alcay S, Toker MB, Onder NT, Gokce E. Royal jelly supplemented soybean lecithin-based extenders improve post-thaw quality and incubation resilience of goat spermatozoa. Cryobiology. 2017;74:81-85. doi:10.1016/j.cryobiol.2016.11.011
  • Taylor MA, Guzmán-Novoa E, Morfin N, Buhr MM. Improving viability of cryopreserved honey bee (Apis mellifera L.) sperm with selected diluents, cryoprotectants, and semen dilution ratios. Theriogenology. 2009;72(2):149-159. doi:10.1016/j.theriogenology. 2009.02.012
  • Forouzanfar M, Sharafi M, Hosseini SM, et al. In vitro comparison of egg yolk-based and soybean lecithin-based extenders for cryopreservation of ram semen. Theriogenology. 2010;73(4):480-487. doi:10.1016/j.theriogenology.2009.10.005
  • Vidal AH, Batista AM, da Silva ECB, et al. Soybean lecithin- based extender as an alternative for goat sperm cryopreservation. Small Rumin Res. 2013;109(1):47-51. doi:10.1016/j.smallrumres.2012.07.022
  • Aktar A, Alçay S. L-Karnitin İlave Edilmiş Sulandırıcıların Dondurma-Çözdürme Sonrası Teke Spermasının Spermatolojik Parametleri Üzerine Etkisi. J Res Vet Med. 2022;41(1):37-42. doi:10.30782/jrvm.999299
  • Andò S, Aquila S. Arguments raised by the recent discovery that insulin and leptin are expressed in and secreted by human ejaculated spermatozoa. Mol Cell Endocrinol. 2005;245(1-2):1-6. doi:10.1016/j. mce.2005.09.011
  • Pinto SCC, Souza FA, Arruda RP, et al. Impact of adding different concentrations of IGF-I and insulin to the semen extender on bull sperm quality post-cryopreservation. Arq Bras Med Vet e Zootec. 2023;75(5):771- 786. doi:10.1590/1678-4162-12946
  • van Tilburg MF, de Oliveira RV, Gadelha CRF, et al. Effects of insulin on sperm cell quality in ram semen cooled at 5°C. Semin Agrar. 2021;42(6):3802-3812. doi:10.5433/1679-0359.2021v42n6Supl2p3803
  • Önder NT, Gökdemir T, Kiliç MC, et al. Insulin and Bull Sperm Interactions During Cryopreservation. Kafkas Univ Vet Fak Derg. 2023;29(4):401-405. doi:10.9775/kvfd.2023.29623
  • Aquila S, Gentile M, Middea E, Catalano S, Andò S. Autocrine regulation of insulin secretion in human ejaculated spermatozoa. Endocrinology. 2005;146(2):552- 557. doi:10.1210/en.2004-1252
  • Lampiao F, Du Plessis SS. Insulin and leptin enhance human sperm motility, acrosome reaction and nitric oxide production. Asian J Androl. 2008;10(5):799-807. doi:10.1111/j.1745-7262.2008.00421.x
  • Shokri S, Ebrahimi SM, Ziaeipour S, Nejatbakhsh R.Effect of insulin on functional parameters of human cryopreserved sperms. Cryobiology. 2019;87(February):68-73. doi:10.1016/j.cryobiol.2019.02.002
  • Alcay S, Cakmak S, Cakmak I, et al. L-carnitine supplemented extenders improve post-thawing quality of honey bee drone (Apis mellifera) spermatozoa[1]. Kafkas Univ Vet Fak Derg. 2021;27(4):489-493. doi:10.9775/kvfd.2021.25756
  • Alçay S, Çakmak S, Çakmak İ, et al. Drone semen cryopreservation with protein supplemented TLhepes based extender. Kafkas Univ Vet Fak Derg. 2019;25(4):553-557. doi:10.9775/kvfd.2018.21311
  • Nur Z, Seven Çakmak S, Çakmak İ, et al. Effects of Trehalose Supplementation on Post-Thaw Sperm Quality of Honey Bee Drones. Online J Anim Feed Res. 2020;10(5):191-196. doi:10.51227/ojafr.2020.27
  • Alcay S, Cakmak S, Cakmak I, et al. Successful cryopreservation of honey bee drone spermatozoa with royal jelly supplemented extenders. Cryobiology. 2019;87(October 2018):28-31. doi:10.1016/j.cryobiol. 2019.03.005
  • Wegener J, May T, Kamp G, Bienefeld K. A successful new approach to honeybee semen cryopreservation. Cryobiology. 2014;69(2):236-242. doi:10.1016/j.cryobiol. 2014.07.011
  • Gürler H, Malama E, Heppelmann M, et al. Effects of cryopreservation on sperm viability, synthesis of reactive oxygen species, and DNA damage of bovine sperm. Theriogenology. 2015;86(2):562-571. doi:10.1016/j. theriogenology.2016.02.007
  • Nur Z, Zik B, Ustuner B, Sagirkaya H, Ozguden CG. Effects of different cryoprotective agents on ram sperm morphology and DNAintegrity. Theriogenology. 2010;73(9):1267-1275. doi:10.1016/j.theriogenology. 2009.12.007
  • Alçay S, Toker MB, Gökçe E, Önder NT, Üstüner B, Nur Z. Long term incubation resilience of post-thaw ram semen diluted with lecithin-based extender supplemented with bovine serum albumin. Kafkas Univ Vet Fak Derg. 2019;25(3):291-297. doi:10.9775/kvfd.2018.20843
  • Çakmak İ. Beekeeping and Recent Colony Losses in Turkey. Uludağ Arıcılık Derg. 2016;16(1):31-48. doi:10.31467/uluaricilik.379276
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Arı Sperması (Apis mellifera anatoliaca) ve İnsülin Etkileşiminin Soya Lesitini Temelli Kriyoprezervasyon Sulandırıcılarında Flow Sitometri Olarak incelenmesi

Yıl 2024, Cilt: 43 Sayı: 1, 46 - 54, 02.09.2024
https://doi.org/10.30782/jrvm.1420188

Öz

Gen havuzlarının istenen yönde gelişimine katkıda bulunan üreme kontrolü, spermanın dondurması ile alakalı biyoteknolojik yöntemleri önemli bir noktaya koymaktadır. Sperma dondurmaya yöntelik çalışmalar özellikle memelilerde yoğun olarak çalışılmış olmasına rağmen, arı sperması açısından başlangıç aşamasındadır. İnsülinin seminal plazmada bulunduğu ve çeşitli sperma parametreleri üzerine etkilerinin olduğu güncel çalışmalarla desteklenmiştir. Ayrıca, soya lesitini uzun süredir, özellikle yumurta sarısına alternatif, güvenilir bir kaynak olarak sperma dondurma sulandırıcılarında tercih edilen maddelerden biridir. Bu çalışma, soya lesitini (%2) temelli arı sperma dondurma sulandırıcılarına farklı dozlarda insülin (5, 10 ve 15 IU dozda) eklenmesi sonucunda elde edilecek sonuçların, flow sitometri veya mikroskopik incelemeler aracılığıyla önemli sperma parametreleri üzerindeki etkileri değerlendirmek amacıyla gerçekleştirişmiştir. Cinsel olgunluğa ulaşmış arılardan elde edilen örnekler bireysel farklılığı engellemek adına birleştirildi (pooling). Dondurma işlemi, üç doz insülin destekli ve bir insülin içermeyen kontrol grubu olacak şekilde düzenlendi ve yöntem planlandığı şekilde uygulandı. Dondurulup çözülen örnekler; motilite, plazma membran (hipo-osmotik şişme testi) ve DNA bütünlüğü (TUNEL testi) değerlendirmelerine, floresan ataçmanı olan bir faz kontrast mikroskobu kullanılarak gerçekleştirildi. Ayrıca, akrozomal bütünlük (PNA-FITC), nitrik oksit seviyeleri (DAF-2/DA) ve mitokondriyal membran potansiyeli (JC-1), bir flow sitometri cihazı aracılığıyla araştırıldı. Beklendiği gibi, tüm sperma parametreleri dondurma işleminin doğası gereği etkilendi, ancak insülin eklenmiş çalışma grupları, kontrol grubuna kıyasla pozitif yönde bir istatistiki farkı ortaya koyamadı. Bu sonuçlar ışığında, soya lesitini temelli arı sperması dondurma sulandırıcılarında kullanılan insülin takviyesinin tercih edilen dozlarda kullanılmasının, yardımcı biyoteknolojik yöntemler aracılığıyla daha yüksek verimli arı kolonileri elde edilmesi ve arı ürünleri konusunda ülke veriminin istenilen düzeye çıkartılması konusunda olumlu bir etkiye sahip olmadığı tespit edildi.

Proje Numarası

Bursa Uludağ Üniversitesi Bilimsel Araştırma Projeleri Birimi (BAP) - TGA – 2022 - 718

Kaynakça

  • Henry LC, Álvaro D rebolledo, Karen C medina, Jorge V rodríguez, Francisco E valente. Effect of egg yolk on sperm cryopreservation of drone (Apis mellifera). Abanico Vet. 2019;9(December):1-11. doi:10.21929/ abavet2019.921
  • Stankus T. A review and bibliography of the literature of honey bee colony collapse disorder: A poorly understood epidemic that Clearly Threatens the successful pollination of billions of dollars of crops in America. J Agric Food Inf. 2008;9(2):115-143. doi:10.1080/10496500802173939
  • Gül A, Şahinler N, Onal AG, Hopkins BK, Sheppard WS. Effects of diluents and plasma on honey bee (Apis mellifera L.) drone frozen-thawed semen fertility. Theriogenology. 2017;101:109-113. doi:10.1016/j. theriogenology.2017.06.020
  • Woyke J, Jasinski Z. Influence of Age of Drones on the Results of Instrumental Insemination of Honeybee Queens. Apidologie. 1978;9(3):203-212. doi:10.1051/ apido:19780304
  • Cobey SW. Comparison studies of instrumentally inseminated and naturally mated honey bee queens and factors affecting their performance. Apidologie. 2007;38(4):390-410. doi:10.1051/apido:2007029
  • Alçay S, Üstüner B, Çakmak İ, Çakmak S, Nur Z. Effects of Various Cryoprotective Agents on Post-Thaw Drone Semen Quality. Kafkas Univ Vet Fak Derg. 2015;21(1):31-35. doi:10.9775/kvfd.2014.11515
  • Almadaly EA, Tawfik FS, El-Kon II, Heleil BA, Fattouh ESM. Effect of different cryoprotectants on the post-thaw sperm characteristics and in vivo fertility of buffalo (Bubalus bubalis) bull semen. Slov Vet Res. 2019;56(February):541-551. doi:10.26873/SVR-792-2019
  • Tonieto RA, Goularte KL, Gastal GDA, Schiavon RS, Deschamps JC, Lucia T. Cryoprotectant effect of trehalose and low-density lipoprotein in extenders for frozen ram semen. Small Rumin Res. 2010;93(2-3):206- 209. doi:10.1016/j.smallrumres.2010.05.003
  • Abdelkader F Ben, Kairo G, Bonnet M, Barbouche N, Belzunces LP, Brunet JL. Effects of clothianidin on antioxidant enzyme activities and malondialdehyde level in honey bee drone semen. J Apic Res. 2019;58(5):740- 745. doi:10.1080/00218839.2019.1655182
  • Kulaksiz R, Daskin AD. In vitro evaluation of Saanen buck semen frozen in different extenders supplemented with various antioxidants. Ankara Üniv Vet Fak Derg. 2010;57:151-156. doi:10.1501/Vetfak_0000002369
  • Borsuk G, Kozłowska M, Anusiewicz M, Paleolog J. Nosema ceranae changes semen characteristics and damages sperm DNA in honeybee drones. Invertebr Surviv J. 2018;15:197-202.
  • Ciereszko A, Wilde J, Dietrich GJ, et al. Sperm parameters of honeybee drones exposed to imidacloprid. Apidologie. 2017;48(2):211-222. doi:10.1007/s13592- 016-0466-2
  • Toker MB, Alcay S, Gokce E, Ustuner B. Cryopreservation of ram semen with antioxidant supplemented soybean lecithin-based extenders and impacts on incubation resilience. Cryobiology. 2016;72(3):205-209. doi:10.1016/j.cryobiol.2016.05.001
  • Alcay S, Toker MB, Onder NT, Gokce E. Royal jelly supplemented soybean lecithin-based extenders improve post-thaw quality and incubation resilience of goat spermatozoa. Cryobiology. 2017;74:81-85. doi:10.1016/j.cryobiol.2016.11.011
  • Taylor MA, Guzmán-Novoa E, Morfin N, Buhr MM. Improving viability of cryopreserved honey bee (Apis mellifera L.) sperm with selected diluents, cryoprotectants, and semen dilution ratios. Theriogenology. 2009;72(2):149-159. doi:10.1016/j.theriogenology. 2009.02.012
  • Forouzanfar M, Sharafi M, Hosseini SM, et al. In vitro comparison of egg yolk-based and soybean lecithin-based extenders for cryopreservation of ram semen. Theriogenology. 2010;73(4):480-487. doi:10.1016/j.theriogenology.2009.10.005
  • Vidal AH, Batista AM, da Silva ECB, et al. Soybean lecithin- based extender as an alternative for goat sperm cryopreservation. Small Rumin Res. 2013;109(1):47-51. doi:10.1016/j.smallrumres.2012.07.022
  • Aktar A, Alçay S. L-Karnitin İlave Edilmiş Sulandırıcıların Dondurma-Çözdürme Sonrası Teke Spermasının Spermatolojik Parametleri Üzerine Etkisi. J Res Vet Med. 2022;41(1):37-42. doi:10.30782/jrvm.999299
  • Andò S, Aquila S. Arguments raised by the recent discovery that insulin and leptin are expressed in and secreted by human ejaculated spermatozoa. Mol Cell Endocrinol. 2005;245(1-2):1-6. doi:10.1016/j. mce.2005.09.011
  • Pinto SCC, Souza FA, Arruda RP, et al. Impact of adding different concentrations of IGF-I and insulin to the semen extender on bull sperm quality post-cryopreservation. Arq Bras Med Vet e Zootec. 2023;75(5):771- 786. doi:10.1590/1678-4162-12946
  • van Tilburg MF, de Oliveira RV, Gadelha CRF, et al. Effects of insulin on sperm cell quality in ram semen cooled at 5°C. Semin Agrar. 2021;42(6):3802-3812. doi:10.5433/1679-0359.2021v42n6Supl2p3803
  • Önder NT, Gökdemir T, Kiliç MC, et al. Insulin and Bull Sperm Interactions During Cryopreservation. Kafkas Univ Vet Fak Derg. 2023;29(4):401-405. doi:10.9775/kvfd.2023.29623
  • Aquila S, Gentile M, Middea E, Catalano S, Andò S. Autocrine regulation of insulin secretion in human ejaculated spermatozoa. Endocrinology. 2005;146(2):552- 557. doi:10.1210/en.2004-1252
  • Lampiao F, Du Plessis SS. Insulin and leptin enhance human sperm motility, acrosome reaction and nitric oxide production. Asian J Androl. 2008;10(5):799-807. doi:10.1111/j.1745-7262.2008.00421.x
  • Shokri S, Ebrahimi SM, Ziaeipour S, Nejatbakhsh R.Effect of insulin on functional parameters of human cryopreserved sperms. Cryobiology. 2019;87(February):68-73. doi:10.1016/j.cryobiol.2019.02.002
  • Alcay S, Cakmak S, Cakmak I, et al. L-carnitine supplemented extenders improve post-thawing quality of honey bee drone (Apis mellifera) spermatozoa[1]. Kafkas Univ Vet Fak Derg. 2021;27(4):489-493. doi:10.9775/kvfd.2021.25756
  • Alçay S, Çakmak S, Çakmak İ, et al. Drone semen cryopreservation with protein supplemented TLhepes based extender. Kafkas Univ Vet Fak Derg. 2019;25(4):553-557. doi:10.9775/kvfd.2018.21311
  • Nur Z, Seven Çakmak S, Çakmak İ, et al. Effects of Trehalose Supplementation on Post-Thaw Sperm Quality of Honey Bee Drones. Online J Anim Feed Res. 2020;10(5):191-196. doi:10.51227/ojafr.2020.27
  • Alcay S, Cakmak S, Cakmak I, et al. Successful cryopreservation of honey bee drone spermatozoa with royal jelly supplemented extenders. Cryobiology. 2019;87(October 2018):28-31. doi:10.1016/j.cryobiol. 2019.03.005
  • Wegener J, May T, Kamp G, Bienefeld K. A successful new approach to honeybee semen cryopreservation. Cryobiology. 2014;69(2):236-242. doi:10.1016/j.cryobiol. 2014.07.011
  • Gürler H, Malama E, Heppelmann M, et al. Effects of cryopreservation on sperm viability, synthesis of reactive oxygen species, and DNA damage of bovine sperm. Theriogenology. 2015;86(2):562-571. doi:10.1016/j. theriogenology.2016.02.007
  • Nur Z, Zik B, Ustuner B, Sagirkaya H, Ozguden CG. Effects of different cryoprotective agents on ram sperm morphology and DNAintegrity. Theriogenology. 2010;73(9):1267-1275. doi:10.1016/j.theriogenology. 2009.12.007
  • Alçay S, Toker MB, Gökçe E, Önder NT, Üstüner B, Nur Z. Long term incubation resilience of post-thaw ram semen diluted with lecithin-based extender supplemented with bovine serum albumin. Kafkas Univ Vet Fak Derg. 2019;25(3):291-297. doi:10.9775/kvfd.2018.20843
  • Çakmak İ. Beekeeping and Recent Colony Losses in Turkey. Uludağ Arıcılık Derg. 2016;16(1):31-48. doi:10.31467/uluaricilik.379276
  • Yániz JL, Silvestre MA, Santolaria P. Sperm quality assessment in honey bee drones. Biology (Basel). 2020;9(7):1-16. doi:10.3390/biology9070174
  • Baumgard LH, Hausman GJ, Sanz Fernandez MV Insulin: Pancreatic secretion and adipocyte regulation. Domest Anim Endocrinol. 2016;54:76-84. doi:10.1016/j.domaniend.2015.07.001
  • Önder NT, Gökdemir T, Kılıç MC, Yıldız S, Öztürkler Y. Effects of Insulin Lispro on Ram Semen Cryopreservation. Kocatepe Vet J. 2023;10:204-212. doi:10.30607/kvj.1347383
  • Vazquez JM, Martinez EA, Martinez P, Garcia-Artiga C, Roca J. Hypoosmotic swelling of boar spermatozoa compared to other methods for analysing the sperm membrane. Theriogenology. 1997;47(4):913-922. doi:10.1016/S0093-691X(97)00046-0
  • Fraser L, Gorszczaruk K, Strzezek J. Relationship between motility and membrane integrity of boar spermatozoa in media varying in osmolality. Reprod Domest Anim. 2001;36(6):325-329. doi:10.1046/ j.1439-0531.2001.00310.x
  • Yániz J, Palacín I, Vicente-Fiel S, Gosalvez J, López-Fernández C, Santolaria P. Comparison of membrane-permeant fluorescent probes for sperm viability assessment in the ram. Reprod Domest Anim. 2013;48(4):598-603. doi:10.1111/rda.12132
  • Önder NT, Gökdemir T, Kiliç MC, Yildiz S, Öztürkler Y. Effects of Insulin Lispro on Ram Semen During Cryopreservation. Kocatepe Vet J. 2017;10:204-212. doi:10.30607/kvj.
  • Cappello AR, Guido C, Santoro A, et al. The mitochondrial citrate carrier (CIC) is present and regulates insulin secretion by human male gamete. Endocrinology. 2012;153(4):1743-1754. doi:10.1210/en.2011-1562
  • Watson PF. The causes of reduced fertility with cryopreserved semen. Anim Reprod Sci. 2000;60-61:481- 492. doi:10.1016/S0378-4320(00)00099-3
  • Neild DM, Gadella BM, Chaves MG, Miragaya MH, Colenbrander B, Agüero A. Membrane changes during different stages of a freeze-thaw protocol for equine semen cryopreservation. Theriogenology. 2003;59(8):1693-1705. doi:10.1016/S0093-691X(02)01231-1
  • Ustuner B, Gokce E, Toker MB, et al. Effect of sperm pooling with seminal plasma collected in breeding or nonbreeding season on Saanen goat sperm cryosurvival. Andrologia. 2018;50(4). doi:10.1111/and.12968
  • Salamon S, Maxwell WMC. Frozen storage of ram semen I. Processing, freezing, thawing and fertility after cervical insemination. Anim Reprod Sci. 1995;37(3-4):185-249. doi:10.1016/0378-4320(94)01327-I
  • Sharafi M, Zhandi M, Shahverdi A, Shakeri M. Beneficial effects of nitric oxide induced mild oxidative stress on post-thawed bull semen quality. Int J Fertil Steril. 2015;9(2):230-237.
  • Ball BA. Oxidative stress, osmotic stress and apoptosis: Impacts on sperm function and preservation inthe horse. Anim Reprod Sci. 2008;107(3-4):257-267. doi:10.1016/j.anireprosci.2008.04.014
  • Ortega Ferrusola C, González Fernandez L, Macías García B, et al. Effect of cryopreservation on nitric oxide production by stallion spermatozoa. Biol Reprod.2009;81(6):1106-1111. doi:10.1095/biolreprod.109.078220
  • Herrero MB, Gagnon C. Nitric oxide: A novel mediator of sperm function. J Androl. 2001;22(3):349-356. doi:10.1002/j.1939-4640.2001.tb02188.x
  • Rosselli M, Keller PJ, Dubey RK. Role of nitric oxide in the biology, physiology and pathophysiology of reproduction. Hum Reprod Update. 1998;4(1):3-24. doi:10.1093/humupd/4.1.3
  • Monnier L, Hanefeld M, Schnell O, Colette C, Owens D. Insulin and atherosclerosis: How are they related? Diabetes Metab. 2013;39(2):111-117. doi:10.1016/j.diabet. 2013.02.001
  • López-Fernández C, Fernández JL, Gosálbez A, et al. Dynamics of sperm DNA fragmentation in domestic animals. III. Ram. Theriogenology. 2008;70(6):898-908. doi:10.1016/j.theriogenology.2008.04.055
  • Corona M, Velarde RA, Remolina S, et al. Vitellogenin, juvenile hormone, insulin signaling, and queen honey bee longevity. Proc Natl Acad Sci U S A. 2007;104(17):7128-7133. doi:10.1073/pnas.0701909104
Toplam 54 adet kaynakça vardır.

Ayrıntılar

Birincil Dil Türkçe
Konular Veteriner Bilimleri (Diğer)
Bölüm Research Articles
Yazarlar

Mehmed Berk Toker 0000-0003-4033-9749

Ahmet Aktar 0000-0002-2975-2594

Selvinar Seven-çakmak 0000-0002-2674-0731

İbrahim Çakmak 0000-0002-8000-5770

Mustafa Akkaşoğlu 0000-0001-6876-0821

Selim Alçay 0000-0002-2472-8157

Proje Numarası Bursa Uludağ Üniversitesi Bilimsel Araştırma Projeleri Birimi (BAP) - TGA – 2022 - 718
Yayımlanma Tarihi 2 Eylül 2024
Gönderilme Tarihi 17 Ocak 2024
Kabul Tarihi 27 Mart 2024
Yayımlandığı Sayı Yıl 2024 Cilt: 43 Sayı: 1

Kaynak Göster

APA Toker, M. B., Aktar, A., Seven-çakmak, S., Çakmak, İ., vd. (2024). Arı Sperması (Apis mellifera anatoliaca) ve İnsülin Etkileşiminin Soya Lesitini Temelli Kriyoprezervasyon Sulandırıcılarında Flow Sitometri Olarak incelenmesi. Journal of Research in Veterinary Medicine, 43(1), 46-54. https://doi.org/10.30782/jrvm.1420188
AMA Toker MB, Aktar A, Seven-çakmak S, Çakmak İ, Akkaşoğlu M, Alçay S. Arı Sperması (Apis mellifera anatoliaca) ve İnsülin Etkileşiminin Soya Lesitini Temelli Kriyoprezervasyon Sulandırıcılarında Flow Sitometri Olarak incelenmesi. J Res Vet Med. Eylül 2024;43(1):46-54. doi:10.30782/jrvm.1420188
Chicago Toker, Mehmed Berk, Ahmet Aktar, Selvinar Seven-çakmak, İbrahim Çakmak, Mustafa Akkaşoğlu, ve Selim Alçay. “Arı Sperması (Apis Mellifera Anatoliaca) Ve İnsülin Etkileşiminin Soya Lesitini Temelli Kriyoprezervasyon Sulandırıcılarında Flow Sitometri Olarak Incelenmesi”. Journal of Research in Veterinary Medicine 43, sy. 1 (Eylül 2024): 46-54. https://doi.org/10.30782/jrvm.1420188.
EndNote Toker MB, Aktar A, Seven-çakmak S, Çakmak İ, Akkaşoğlu M, Alçay S (01 Eylül 2024) Arı Sperması (Apis mellifera anatoliaca) ve İnsülin Etkileşiminin Soya Lesitini Temelli Kriyoprezervasyon Sulandırıcılarında Flow Sitometri Olarak incelenmesi. Journal of Research in Veterinary Medicine 43 1 46–54.
IEEE M. B. Toker, A. Aktar, S. Seven-çakmak, İ. Çakmak, M. Akkaşoğlu, ve S. Alçay, “Arı Sperması (Apis mellifera anatoliaca) ve İnsülin Etkileşiminin Soya Lesitini Temelli Kriyoprezervasyon Sulandırıcılarında Flow Sitometri Olarak incelenmesi”, J Res Vet Med, c. 43, sy. 1, ss. 46–54, 2024, doi: 10.30782/jrvm.1420188.
ISNAD Toker, Mehmed Berk vd. “Arı Sperması (Apis Mellifera Anatoliaca) Ve İnsülin Etkileşiminin Soya Lesitini Temelli Kriyoprezervasyon Sulandırıcılarında Flow Sitometri Olarak Incelenmesi”. Journal of Research in Veterinary Medicine 43/1 (Eylül 2024), 46-54. https://doi.org/10.30782/jrvm.1420188.
JAMA Toker MB, Aktar A, Seven-çakmak S, Çakmak İ, Akkaşoğlu M, Alçay S. Arı Sperması (Apis mellifera anatoliaca) ve İnsülin Etkileşiminin Soya Lesitini Temelli Kriyoprezervasyon Sulandırıcılarında Flow Sitometri Olarak incelenmesi. J Res Vet Med. 2024;43:46–54.
MLA Toker, Mehmed Berk vd. “Arı Sperması (Apis Mellifera Anatoliaca) Ve İnsülin Etkileşiminin Soya Lesitini Temelli Kriyoprezervasyon Sulandırıcılarında Flow Sitometri Olarak Incelenmesi”. Journal of Research in Veterinary Medicine, c. 43, sy. 1, 2024, ss. 46-54, doi:10.30782/jrvm.1420188.
Vancouver Toker MB, Aktar A, Seven-çakmak S, Çakmak İ, Akkaşoğlu M, Alçay S. Arı Sperması (Apis mellifera anatoliaca) ve İnsülin Etkileşiminin Soya Lesitini Temelli Kriyoprezervasyon Sulandırıcılarında Flow Sitometri Olarak incelenmesi. J Res Vet Med. 2024;43(1):46-54.