Damage assessment of the equine sperm membranes by fluorimetric technique

Author(s)

Carvalho Celeghini, Eneiva Carla ^[1] ; Cesar de Andrade, Andre Furugen ^[1] ; Raphael, Claudia Fernandes ^[1] ; Nascimento, Juliana ^[1] ; Ticianelli, Janahi Souza ^[1] ; de Arruda, Rubens Paes ^[1]

Affiliation

^[1] Univ Sao Paulo, Lab Biotecnol Semen & Androl, Dept Anim Reprod, Fac Med Vet & Zootecn, BR-13635900 Pirassununga, SP - Brazil

• Document type: Journal article
• Source: Brazilian Archives of Biology and Technology; v. 53, n. 6, p. 1285-1292, NOV-DEC 2010.
• Web of Science Citations: 6

Abstract

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. (AU)