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Changes in binding of lectins to epididymal, ejaculated, and capacitated spermatozoa of the rhesus monkey
D Navaneetham1, P Sivashanmugam, M Rajalakshmi
1Department of Reproductive Biology, All India Institute of Medical Science, New Delhi, India.
Background:
The present study was undertaken to evaluate the changes in rhesus monkey sperm surface glycoconjugates during maturation, ejaculation, and capacitation in order to provide background information that would help in evaluating adverse effects, if any, caused by the use of contraceptive agents.
Methods:
Adult sexually mature rhesus monkeys were castrated under ketamine anaesthesia. Percoll purified sperm from different epididymal segments and motile-ejaculated spermatozoa prior to and following in vitro capacitation were exposed to FITC-labeled lectins. Live or sperm prefixed with paraformaldehyde/alcohol were used. Quantitative analysis of changes in lectin binding was done immunoassay.
Results:
The majority of live spermatozoa did not show binding of Con A and PNA, whereas uniform labeling of WGA to sperm from corpus epididymidis onward was seen. Unfixed spermatozoa showed marked variation in the pattern of lectin binding. The majority of spermatozoa fixed with paraformaldehyde or alcohol showed maximum lectin labeling over the acrosome, but the postacrosomal area invariably did not bind the lectins. Major changes in the localization of Con A, PNA, or WGA to prefixed spermatozoa were not seen during epididymal transit and in the ejaculate. In capacitated acrosome-reacted spermatozoa, Con A, PNA, and WGA were localized mainly in the equatorial segment. Binding of Con A and PNA was abolished by using lectins preincubated with appropriate inhibitor saccharides. Sperm, exposed to FITC-WGA preincubated with inhibitor sugar, did not show complete inhibition. Quantitative analysis of lectin binding by immunoassay showed increase in binding of lectins during epididymal transit with maximum binding in sperm from the corpus epididymidis.
Conclusions:
The variations in lectin labeling of live spermatozoa could be due to redistribution of sperm surface sugars or membrane damage. The changes in lectin labeling during maturation and capacitation may be associated with their role in ovum recognition and fusion.