ULTRASTRUCTURAL ANALYSIS OF RAT SPERMATOZOA FOLLOWING TREATMENT WITH CO-POLYMER STYRENE MALEIC ANHYDRIDE (SMA) IN VITRO
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Date
2019-12
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Abstract
Administration of co-polymer SMA dissolved in the vehicle Dimethyl Sulfoxide (DMSO), into the lumen
of the vas deferens is a recent and very effective vas deferens-based fertility control procedure that blocks the
lumen of the vas deferens and also impairs the fertilizing ability of the spermatozoa that pass in the vicinity
of the lumen because of its electrical charge and pH-lowering action (Lohiya et al., 1998). In the present
study, the micro structural alterations caused by treatment of rat spermatozoa with Styrene Maleic Anhydride
(SMA) in vitro were analysed by Atomic Force Microscopy (AFM) and Scanning Electron Microscopy
(SEM). Spermatozoa recovered by mincing method from sacrificed adult male Wistar rats were utilized
for the study. The recovered sperms were microscopically examined for their motility, diluted with TRIS
buffer and incubated for 60 minutes with co-polymer SMA prepared by Reversible Addition-Fragmentation
chain-Transfer (RAFT) polymerization technique@ 2 mg in 100 μl of DMSO. Washed epididymal sperms
were fixed with 2.5% Glutaraldehyde in PBS (pH 7.4) for 30 min at 37°C and immobilized on 0.01% Poly-
L-Lysine coated slides prior to analysis. Tapping mode AFM of untreated rat spermatozoa showed regularacrosome, mid-piece, post-acrosomal segment and flagellum that were clearly visible due to local height
variations whereas SMA treated rat epididymal sperms revealed disintegrated surface topology with complete
loss of smoothness. For SEM, a thin film of spermatozoa was smeared on a SEM stub with silver paint, airdried,
sputter-coated with gold and observed under scanning electron microscope. Untreated spermatozoa
had regular smooth sperm head with distinct acrosome, principle piece and flagellum whereas SMA treated
spermatozoa revealed distorted acrosome. Hence, it is concluded that SMA can be a great breakthrough in
canine contraception when injected through intravasal route which is already in phase III clinical trials in
humans.
Description
TNV_35thVCRI_CA_Dec-2019_EPWO-27
Keywords
Veterinary Science