Abstract
In the present study, it was aimed to evaluate the effects of semen extender with added insulin on ram semen at the post-thaw stage. Semen was collected from 5 Tuj rams five times with an electro-ejaculator. Experimental groups were designated as control group and groups containing 15 IU and 20 IU insulin lispro added semen extenders. At the post-thaw stage, motility and plasma membrane integrity were evaluated by microscopy. Acrosome integrity (Fitc-Peanut Agglutinin) and mitochondrial membrane potential (Rhodamine 123) were assessed by dual staining with propidium iodide (PI) using flow cytometry. It was found that motility and plasma membrane functional integrity were better preserved in the control group (p<0.05). Acrosome integrity results were similar between the control and 15 IU insulin groups (p>0.05), but acrosome integrity was negatively affected in the 20 IU insulin group (p<0.05). Mitochondrial membrane potential was higher in the group containing 20 IU insulin (p<0.05). As a result, it was thought that energy metabolism was stimulated in ram semen frozen with insulin-added extenders, and the semen was better preserved than the control group.
Keywords
Supporting Institution
Kafkas University Scientific Research Projects Unit
Project Number
2022-TS-55
References
- Aitken RJ, Curry BJ, Shokri S, Pujianto DA, Gavriliouk D, Gibb Z, Whiting S, Connaughton HS, Nixon B, Salamonsen LA, Baker MA. Evidence that extrapancreatic insulin production is involved in the mediation of sperm survival. Mol Cell Endocrinol. 2021; 526: 111193.
- Alamo A, De Luc C, Mongioì LM, Barbagallo F, Cannarella R, La Vignera S, Calogero AE, Condorelli RA. Mitochondrial membrane potential predicts 4-hour sperm motility. Biomedicines. 2020; 8 (7): 196.
- Alcay S, Aktar A, Koca D, Kilic MA, Akkasoglu M, Sagirkaya H. Positive effect of autologous platelet rich plasma on Saanen buck semen cryopreservation in non-breeding season. Cryobiology. 2021; 103: 45-48.
- Alçay S, Cakmak S, Cakmak I, Aktar A, Ustuner B, Yilmaz M, Duman M, Ozkan H, Akkaya Y, Sağırkaya H, Nur, Z. Honey bee drone (Apis mellifera) sperm cryopreservation with rainbow trout seminal plasma supplemented extenders. J Hellenic Vet Med Soc. 2022; 73 (1): 3751-3756.
- Alcay S, Gokce E, Toker MB, Onder NT, Ustuner B, Uzabacı E, Gul Z, Cavus S. Freeze-dried egg yolk based extenders containing various antioxidants improve post-thawing quality and incubation resilience of goat spermatozoa. Cryobiology. 2016; 72 (3): 269-273.
- Aquila S, Giordano F, Guido C, Rago V, Carpino A. Insulin affects sperm capacity in pig through nitric oxide. Asian J Androl. 2013; 15 (6): 835-837.
- Bruning JC, Gautam D, Burks DJ, Gillette J, Schubert M, Orban PC, Klein R, Krone W, Müller-Wieland D, Kahn CR. Role of brain insulin receptor in control of body weight and reproduction. Science. 2000; 289 (5487): 2122-2125.
- Dias TR, Alves MG, Silva BM, Oliveira PF. Sperm glucose transport and metabolism in diabetic individuals. Mol Cell Endocrinol. 2014; 396 (1-2): 37-45.
- Gürler H, Malama E, Heppelmann M, Calisici O, Leiding C, Kastelic JP, Bollwein H. Effects of cryopreservation on sperm viability, synthesis of reactive oxygen species, and DNA damage of bovine sperm. Theriogenology. 2016; 86 (2): 562-571.
- Holleman F, Hoekstra JB. Insulin lispro. N Engl J Med, 1997; 337 (3): 176-183.
- Karamitsos DT. The story of insulin discovery. Diabetes Res Clin Pract. 2011; 93: 2-8.
- Koyunlu A. Spor ve Egzersiz Metabolizmasına Güncel Bakış, In: İnsülin Metabolizması ve Egzersiz İlişkisi, Ed; Pancar Z, 1st Ed., İstanbul, 2023; pp. 73-89.
- 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.
- Lunze K, Singh T, Walter M, Brendel MD, Leonhardt S. Blood glucose control algorithms for type 1 diabetic patients: A methodological review. Biomed Signal Process Control. 2013; 8 (2): 107-119.
- Noble SL, Johnston E, Walton B. Insulin lispro: a fast-acting insulin analog. American Family Physician, 1998; 57 (2): 279-286.
- Onder NT, Alcay S, Nur Z. Effects of alpha‐lipoic acid on ram semen cryopreservation and post‐thaw life span. Andrologia. 2022; 54 (1): e14249.
- Silvestroni L, Modesti A, Sartori C. Insulin-sperm interaction: effects on plasma membrane and binding to acrosome. Arch Androl. 1992; 28 (3): 201-211.
- van Tilburg MF, de Oliveira RV, Gadelha CRF, Fagundes B, Dias AJB, Quirino CR, Silva JSF. Effects of insulin on sperm cell quality in ram semen cooled at 5° C. Semina Cien Agrar, 2021; 42 (6): 3803-3812.
- 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.
- Wang X, Sharma RK, Gupta A, George V, Thomas Jr AJ, Falcone T, Agarwal A. Alterations in mitochondria membrane potential and oxidative stress in infertile men: a prospective observational study. Fertil Steril. 2003; 80: 844-850.
- Weerakoon, W. W. P. N., Sakase, M., Kohama, N., & Kawate, N. Plasma estradiol-17β, cortisol, and insulin concentrations and serum biochemical parameters surrounding puberty in Japanese Black beef bulls with normal and abnormal semen. Theriogenology. 2020; 148: 18-26.
- Yildiz S, Öztürkler Y, Ari UC, Lehimcioglu NC, Atakisi E, Kulaksiz R. The effects of l-ergothioneine, n-acetylcystein and cystein on freezing of ram semen. Kafkas Univ Vet Fak Derg. 2015; 21 (1): 81-86.
Abstract
Sunulan çalışmada, insülin ilave edilen sperma sulandırıcısının koç sperması üzerindeki etkilerininin çözdürme sonrası aşamada değerlendirilmesi amaçlanmıştır. Tuj ırkı 5 adet koçtan beş kez elektro-ejakülatör ile sperma alındı. Deneme grupları, kontrol grubu, 15 IU ve 20 IU insulin lispro ilave edilen sperma sulandırıcıları ile oluşturuldu. Çözdürme sonrası, motilite ve plazma membran bütünlüğü mikroskopi ile değerlendirildi. Akrozom bütünlüğü (Fitc-Peanut Agglutinin) ve mitokondrial membran potansiyeli (Rhodamine 123) akış sitometrisi kullanılarak, propidium iodide (PI) ile ikili boyama yapılarak değerlendirildi. Motilite ve plazma membran fonksiyonel bütünlüğünün kontrol grubuna göre daha iyi korunduğu bulundu (p<0.05). Akrozom bütünlüğü sonuçları kontrol ve 15 IU insülin grubu arasında benzer olduğu (p>0.05), ancak 20 IU insulin grubunda akrozom bütünlüğünün olumsuz etkilediği bulundu (p<0.05). Mitokondrial membran potansiyeli, 20 IU insulin içeren grupta daha yüksek bulundu (p<0.05). Sonuç olarak, insulin ilave edilmiş sulandırıcılar ile dondurulan koç spermasında enerji metabolizmasının uyarıldığı ve spermanın kontrol grubuna göre daha iyi korunduğu düşünülmüştür.
Keywords
Project Number
2022-TS-55
References
- Aitken RJ, Curry BJ, Shokri S, Pujianto DA, Gavriliouk D, Gibb Z, Whiting S, Connaughton HS, Nixon B, Salamonsen LA, Baker MA. Evidence that extrapancreatic insulin production is involved in the mediation of sperm survival. Mol Cell Endocrinol. 2021; 526: 111193.
- Alamo A, De Luc C, Mongioì LM, Barbagallo F, Cannarella R, La Vignera S, Calogero AE, Condorelli RA. Mitochondrial membrane potential predicts 4-hour sperm motility. Biomedicines. 2020; 8 (7): 196.
- Alcay S, Aktar A, Koca D, Kilic MA, Akkasoglu M, Sagirkaya H. Positive effect of autologous platelet rich plasma on Saanen buck semen cryopreservation in non-breeding season. Cryobiology. 2021; 103: 45-48.
- Alçay S, Cakmak S, Cakmak I, Aktar A, Ustuner B, Yilmaz M, Duman M, Ozkan H, Akkaya Y, Sağırkaya H, Nur, Z. Honey bee drone (Apis mellifera) sperm cryopreservation with rainbow trout seminal plasma supplemented extenders. J Hellenic Vet Med Soc. 2022; 73 (1): 3751-3756.
- Alcay S, Gokce E, Toker MB, Onder NT, Ustuner B, Uzabacı E, Gul Z, Cavus S. Freeze-dried egg yolk based extenders containing various antioxidants improve post-thawing quality and incubation resilience of goat spermatozoa. Cryobiology. 2016; 72 (3): 269-273.
- Aquila S, Giordano F, Guido C, Rago V, Carpino A. Insulin affects sperm capacity in pig through nitric oxide. Asian J Androl. 2013; 15 (6): 835-837.
- Bruning JC, Gautam D, Burks DJ, Gillette J, Schubert M, Orban PC, Klein R, Krone W, Müller-Wieland D, Kahn CR. Role of brain insulin receptor in control of body weight and reproduction. Science. 2000; 289 (5487): 2122-2125.
- Dias TR, Alves MG, Silva BM, Oliveira PF. Sperm glucose transport and metabolism in diabetic individuals. Mol Cell Endocrinol. 2014; 396 (1-2): 37-45.
- Gürler H, Malama E, Heppelmann M, Calisici O, Leiding C, Kastelic JP, Bollwein H. Effects of cryopreservation on sperm viability, synthesis of reactive oxygen species, and DNA damage of bovine sperm. Theriogenology. 2016; 86 (2): 562-571.
- Holleman F, Hoekstra JB. Insulin lispro. N Engl J Med, 1997; 337 (3): 176-183.
- Karamitsos DT. The story of insulin discovery. Diabetes Res Clin Pract. 2011; 93: 2-8.
- Koyunlu A. Spor ve Egzersiz Metabolizmasına Güncel Bakış, In: İnsülin Metabolizması ve Egzersiz İlişkisi, Ed; Pancar Z, 1st Ed., İstanbul, 2023; pp. 73-89.
- 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.
- Lunze K, Singh T, Walter M, Brendel MD, Leonhardt S. Blood glucose control algorithms for type 1 diabetic patients: A methodological review. Biomed Signal Process Control. 2013; 8 (2): 107-119.
- Noble SL, Johnston E, Walton B. Insulin lispro: a fast-acting insulin analog. American Family Physician, 1998; 57 (2): 279-286.
- Onder NT, Alcay S, Nur Z. Effects of alpha‐lipoic acid on ram semen cryopreservation and post‐thaw life span. Andrologia. 2022; 54 (1): e14249.
- Silvestroni L, Modesti A, Sartori C. Insulin-sperm interaction: effects on plasma membrane and binding to acrosome. Arch Androl. 1992; 28 (3): 201-211.
- van Tilburg MF, de Oliveira RV, Gadelha CRF, Fagundes B, Dias AJB, Quirino CR, Silva JSF. Effects of insulin on sperm cell quality in ram semen cooled at 5° C. Semina Cien Agrar, 2021; 42 (6): 3803-3812.
- 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.
- Wang X, Sharma RK, Gupta A, George V, Thomas Jr AJ, Falcone T, Agarwal A. Alterations in mitochondria membrane potential and oxidative stress in infertile men: a prospective observational study. Fertil Steril. 2003; 80: 844-850.
- Weerakoon, W. W. P. N., Sakase, M., Kohama, N., & Kawate, N. Plasma estradiol-17β, cortisol, and insulin concentrations and serum biochemical parameters surrounding puberty in Japanese Black beef bulls with normal and abnormal semen. Theriogenology. 2020; 148: 18-26.
- Yildiz S, Öztürkler Y, Ari UC, Lehimcioglu NC, Atakisi E, Kulaksiz R. The effects of l-ergothioneine, n-acetylcystein and cystein on freezing of ram semen. Kafkas Univ Vet Fak Derg. 2015; 21 (1): 81-86.
Details
| Primary Language | English |
|---|---|
| Subjects | Veterinary Sciences (Other) |
| Journal Section | RESEARCH ARTICLE |
| Authors | |
| Project Number | 2022-TS-55 |
| Publication Date | September 30, 2023 |
| Acceptance Date | September 25, 2023 |
| Published in Issue | Year 2023 |
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