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

Mehmed Berk Toker; Ahmet Aktar; Selvinar Seven-çakmak; İbrahim Çakmak; Mustafa Akkaşoğlu; Selim Alçay

Article Details

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

Authors:

Mehmed Berk Toker | Ahmet Aktar | Selvinar Seven-çakmak | İbrahim Çakmak | Mustafa Akkaşoğlu | Selim Alçay

Publisher:

Bursa Uludağ University

Main Topics:

Veterinary Sciences (Other)

Journal:

Journal of Research in Veterinary Medicine Vol.43 Issue 1, pp.46-54

Document Type:

Article

Publication Date:

2/2024

Document Language:

Turkish

Document ID (Org No.):

2001326435

pISSN:

2667-6745

Keywords:

Abstract:

 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-2DA, 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 countrys yield in bee products through auxiliary biotechnological methods as desired.

 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-2DA, 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 countrys yield in bee products through auxiliary biotechnological methods as desired.

 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-2DA, 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 countrys yield in bee products through auxiliary biotechnological methods as desired.

 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-2DA 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.

Article Details

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

Authors:

Mehmed Berk Toker | Ahmet Aktar | Selvinar Seven-çakmak | İbrahim Çakmak | Mustafa Akkaşoğlu | Selim Alçay

Publisher:

Bursa Uludağ University

Main Topics:

Veterinary Sciences (Other)

Journal:

Journal of Research in Veterinary Medicine Vol.43 Issue 1, pp.46-54

Document Type:

Article

Publication Date:

2/2024

Document Language:

Turkish

Document ID (Org No.):

2001326435

pISSN:

2667-6745

Apis mellifera anatoliaca

Drone sperm

Flow-cytometer

Insulin

Soybean lecithin

Sperma cryopreservationApis mellifera anatoliaca

Arı sperması

Flow-sitometri

İnsülin

Soya lesitini

Sperma dondurma

Issues

Article Details

Authors: Mehmed Berk Toker | Ahmet Aktar | Selvinar Seven-çakmak | İbrahim Çakmak | Mustafa Akkaşoğlu | Selim Alçay

Publisher: Bursa Uludağ University

Main Topics: Veterinary Sciences (Other)

Journal: Journal of Research in Veterinary Medicine Vol.43 Issue 1, pp.46-54

Document Type: Article

Publication Date: 2/2024

Document Language: Turkish

Document ID (Org No.): 2001326435

pISSN: 2667-6745

Issues

Authors:

Mehmed Berk Toker | Ahmet Aktar | Selvinar Seven-çakmak | İbrahim Çakmak | Mustafa Akkaşoğlu | Selim Alçay

Publisher:

Bursa Uludağ University

Main Topics:

Veterinary Sciences (Other)

Journal:

Journal of Research in Veterinary Medicine Vol.43 Issue 1, pp.46-54

Document Type:

Article

Publication Date:

2/2024

Document Language:

Turkish

Document ID (Org No.):

2001326435

pISSN:

2667-6745