Ülkü Nur CANBOLAT, İlhami ÇELİK, Rukiye ERDOĞAN

Taze ve dondurulup-çözdürülen boğa sperm akrozomlarının alfa-naftil aseteat esteraz aktivitelerinin belirlenmesi

Amaç: Akrozomal enzimlerin seminal plazmaya sızıp sızmadığının belirlenmesi, suni tohumlama için sperma işlenmesi sırasında oluşan akrozom hasarının öncü bir göstergesi olabilir. Alfa-naftil asetat esteraz enzimi bu amaçla kullanılabilecek olan akrozomal enzimlerden biridir. Bu çalışmada, taze ve dondurup-çözdürülen boğa spermatozoonlarındaki alfa-naftil asetat esteraz enziminin pozitivite oranları ve lokalizasyonunun ışık mikroskobuyla belirlenmesi amaçlanmıştır. Gereç ve Yöntem: Çalışmada materyal olarak 7 boğadan alınan sperma örneği kullanıldı. Bu amaçla, taze ve dondurulup-çözdürülen sperma örneklerinden sürme frotiler hazırlandı. Bu frotilerde alfa-naftil asetat esteraz enzimi sitokimyasal metotla demonstre edildi. Hazırlanan preparatlar ışık mikroskobuyla incelenerek alfa-naftil-asetat esteraz pozitivite oranları, enzimin lokalizasyon ve dağılım tarzı belirlendi. Bulgular: Alfa-naftil asetat esteraz enzimi pozitivitesi, spermatozoonun akrozomunda lokalize olan kırmızımsı-kahve renkli reaksiyon ürünü halinde gözlendi. Bazı hücrelerde enzimatik reaksiyon akrozom zarında dar bir band halinde gözlendi. Taze sperma örneklerinin alfa-naftil asetat esteraz pozitivitesinde belirgin bireysel farklılıklar gözlendi. Enzimatik reaksiyon, hücrelerin tamamında aynı şiddette değildi. Bazı hücrelerdeki reaksiyon nispeten daha zayıftı. Düşük spermatozoon vitalitesi ve motilitesi tesbit edilen taze sperma örneklerinin alfa-naftil asetat esteraz enzim pozitivitesi de düşüktü. Dondurup- çözme işlemi, örneklerin alfa-naftil asetat esteraz pozitivitesinde önemli düşüşe yol açtı. Öneri: Akrozomal alfa-naftil asetat esteraz enzimi, boğa spermasının fertilite potansiyelinin ve dondurma işlemine uygunluğunun değerlendirilmesinde ve önemli bir parametre olarak dikkate alınabileceği sonucuna varılmıştır.

Determination of alpha-napthyl acetate esterase activity of native and frozen-thawed bull sperm acrosome

Aim: The assay of the leakage of acrosomal enzymes into seminal plasmamight provide an early and sensitive evidence for membrane damage ocurredduring freezing of semen for artificial insemination. Alpha-naphthyl acetateesterase is one of the acrosomal enzymes that can be used for this purpose.In the the present study, the positivity rates and localization pattern of alphanaphthylacetate esterase in the spermatozoa of both native and frozen-thawedbull semen samples were determined by light microscopy.Materials and Methods: Semen samples of 7 bull were used as material. Smearswere prepared from native and frozen-thawed semen samples. The enzymewas demonstrated by a cytochemical method. Specimens were observedunder ligth microscope. Positivity rates of alpha-naphtyl acetate eseterasepositivespermatozoa, distribution and localization pattern of the enzymaticreaction were determined.Results: The reddish-brown enzymatic reaction was peculiar to spermatozoonacrosome. In some cells, the enzymatic reaction was observed as a narrowband located on acrosomal membrane. The enzymatic reaction intesity wasnot similar in all cells and was weaker in some cells. The reaction productprimarily located in the acrosome. There were large individual differences inalpha-naphthyl acetate esterase positivity rates of the native sperm samples.The native semen samples with low spermatozoon vitality and motility tendedto display lower of alpha-naphthyl acetate esterase positivity rates. Freeze-thawing process significantly decreased alpha-naphthyl acetate esterasepositivity rates of the samples.Conclusion: Acrosomal alpha-naphtyl acetate esterase might be consideredas a significant parameter in the asssessment of availibility for freezing and todetermine fertility potential of the bull semen.

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