Damage assessment of the equine sperm membranes by fluorimetric technique

Brazilian Archives of Biology and Technology

Eneiva Carla Carvalho Celeghini¹  André Furugen Cesar De Andrade¹  Cláudia Fernandes Raphael¹  Juliana Nascimento¹  Janahi Souza Ticianelli¹  Rubens Paes De Arruda¹ 

[1],Universidade de São Paulo Faculdade de Medicina Veterinária e Zootecnia Departamento de Reprodução AnimalPirassununga SP ,Brasil

关键词: Stallion;    acrosome;    plasma membrane;    mitochondria;    fluorescent probes;   

DOI  :  10.1590/S1516-89132010000600004

来源: SciELO

【 摘 要 】

To validate a practical technique of simultaneous evaluation of the plasma, acrosomal and mitochondrial membranes in equine spermatozoa three fluorescent probes (PI, FITC-PSA and MITO) were associated. Four ejaculates from three stallions (n=12) were diluted in TALP medium and split into 2 aliquots, 1 aliquot was flash frozen in liquid nitrogen to induce damage in cellular membranes. Three treatments were prepared with the following fixed ratios of fresh semen: flash frozen semen: 100:0 (T100), 50:50 (T50), and 0:100 (T0). A 150-µL aliquot of diluted semen of each treatment was added of 2 µL of PI, 2 µL of MITO and 80 µL of FITC-PSA; incubated at 38.5ºC/8 min, and sperm cells were evaluated by epifluorescent microscopy. Based in regression analysis, this could be an efficient and practical technique to assess damage in equine spermatozoa, as it was able to determine the sperm percentage more representative of the potential to fertilize the oocyte.

【 授权许可】

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