Indian Journal of Animal Sciences 71 (6): 519·520, June 200 I In vitro capacitation and acrosome reaction of goat sperms in Tyrode's modified solution* S CHEEMA I, G K SANGHA2 and S S GURA Y A3 Punjab Agricultural University, Ludhiana, Punjab 14i 004 india Received: I June 2000; Accepted: 28 November 2000 ABSTRACT Per cent motility, hyperactive motility (capacitation) and acrosome reaction in goat epididymal spermatozoa during incubation in vitro at 38.5°C in m-TALP (Tyrode's modified solution containing albumin, lactate and pyruvate) medium supplemented with taurine and epinephrine as motility factors plus BSA under liquid paraffin. in a <;;02 inCUbator, were. studied. The per cent capacitation and acrosome reaction can be estimated by the onset of hyPeractive motility and shedding of acrosome reaction under an inverted microscope. The percentage of hyperactive motile spel1\latoz,oa i is maximum. (72.40:!: 1.43) at 4hr of incubation, The per cent acrosome reaction increased progressively with of ± 0.79 at fifth hr of incubation. The results suggested that the goat epididymal spermatozoa can be Induced to undergo capacitation and acrosome reaction /1'1 vitro in a chemically defined medium. Key words: Acrosome reaction, Capacitation, Epididymis, Goat, Sperm and are disti*ct and distinguishable processes which are of fundamental. tance in fertilization of an oocyte by a spermatozoa (Dunbar and O'Rand 1991, Guraya 2000). Capacitation was plished in vitro in various well defined media in several mammalian species ( Bavistor 1986, Guraya 2000). For in vitro fertilization and production of embryos under laboratory conditions, a need for sperm capacitation under in vitro con- dition arises. No such work has been carried out previously in goats which are important for meat and milk production in Asian and African countries. Therefore the present study was undertaken to study the capacitation and acrosome reaction process in goat in vitro. MATERIALS AND METHODS Goat testis along with epididymis were procured from a IOC,I} slaughterhouse, brought inunediately to the laboratory in phosphate buffer saline (pH 7.4). The epididymis was carefully separated out and the epididymal spermatozoa were obtained by incising the cauda portion with a sharp blade. The viscous cauda epididymal spermatozoa were released into normal saline containing glucose 1 mg/ml (pH 7.4) in a polypropylene petri plate and were kept for 30·45 min at 38.5"C. Hyperactive motile spermatozoa were collected by Present address: I Research Associate? Assistant Physiologist (Reproduction),' former Professor .and Head, Department of Zoology. swim up procedure using TALPmerliuJl1J i;e. TYFode's modified sohuion adjllsted i to pH.7.4 . .Bamples'with<motility >80% • were used, for. further experimeptatlolt Finally. the spermatozoa were ittOllbated in T ALP medium contaUiing BSA and motility factors under liquid paraffm in a 002 bator for 6·8hr. An hourly assessment of sperm nwti4ity, hyperactive motility of capacitation (Charactetized by whiplash movement of tail) and acrosome reaction (using dual staining technique of Sidhu et al.1992) was estimated under inverted microscope. RESULTS Goat cauda epididymal spermatozoa, collected in !.lOnna1 saline (pH 7.4) at 38.5°C, when seen under low power (lOx) showed very fast movements with waves and eddies and very high concenb"ation of cells. Only 10% agglutination of goat spermatozoa was observed as most of the sperm were seen moving very fast. Alliquotes of incubated semen samples were examined after each hour for percentage of sperm motility under a phase contrast microscope ( 40 x ): The per cent motility of goat epididymal spermatozoa at different hours of incubation is shown in Table 1. The sperm per cent motility estimated to be 80% at Obr of incubation slightly decreased at 1hr of incubation and then again increased at.2hr of incubation, but' started decreasing thereafter (Table '1). The sperm per cent capacitation. started at 3hr and was assessed by analyzing the number ofhyperactivt: in a zig zag manner showing ; . :' ,:' ' _" - , .;,"",: " I , , "::' ,: ,"