Damage Assessment of the Equine Sperm Membranes by Fluorimetric Technique

作者：Carvalho Celeghini Eneiva Carla; Cesar de Andrade Andre Furugen; Raphael Claudia Fernandes; Nascimento Juliana; Ticianelli Janahi Souza; de Arruda Rubens Paes^*

来源：Brazilian Archives of Biology and Technology, 2010, 53(6): 1285-1292.

DOI：10.1590/S1516-89132010000600004

• Stallion
• acrosome
• plasma membrane
• mitochondria
• fluorescent probes

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-mu L aliquot of diluted semen of each treatment was added of 2 mu L of PI, 2 mu L of MITO and 80 mu L of FITC-PSA; incubated at 38.5 degrees C/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.

• 出版日期2010-12