USE OF PROPIDIUM IOIDE AND PISUM-SATIVUM FOR FAST ASSESSMENT OF ACROSOME INTEGRITY IN GOAT SPERMATOZOA

Lectins are used for studies in mammalian sperm surface and organelles because of their ability to specifically link sugar residua. Fluoresc ein labeled Pisum sativum (PSA) has been used successfully in several mammalian spermatozoa, as a marker of the inner acrosomal membrane but , because of interspecific Variations in membrane constituents, its us e requires a preliminary validation. This study compared the results o btained by staining goat spermatozoa with a combination of propidium i oide (PI) and PSA vs those of Bengal Red. Also the interpretation of t he results achieved were compared with the results obtained by two ext ernal technicians. In Experiment I, goat spermatozoa collected by arti ficial vagina were washed once, immediately after collection, and stai ned with PI/PS A and Bengal Rose. Then, to prepare sperm groups with d ifferent proportions of ii ve and dead sperm, spermatozoa were fractio nated and one fraction was damaged by freezing at -196 degrees C witho ut cryoprotectans. In Experiment 2, semen from different males was fro zen in a Tris-base d extender, and thawed either at 37 degrees C for 3 0 sec or at 65 degrees C for 5 sec, and then stained with PI/PSA. Two independent technicians assessed the results of both series of experim ents. Percentage transformations, media comparison and correlation wer e obtained by using the Systat(R) computing package. The results showe d that PI/PSA and Bengal Rose gave similar and positively correlated r esults and that both technicians gave similar interpretations (r = 0.9 8). It can be concluded that PI/PSA is a useful staining for fast asse ssment of goat spermatozoa.