Analysis of sperm carbohydrate recognition molecules involved in fertilization

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It has been shown that a multiple carbohydrate recognition system is working in the gamete interaction. In this project, carbohydrate recognition molecules on boar sperm involved in fertilization were investigated and the following results were obtained.1) Cytochemical analysis using multivalent oligosaccharide probes indicated that two kinds of molecules, one recognizing the N-acetyllactosamine structure and the other recognizing the Le^x structure, are expressed on boar sperm heads.2) These carbohydrate recognition molecules are detected more strongly on capacitated sperm than on uncapacitaed sperm, but not detected on acrosome-reacted sperm.3) When the detergent-solubilized plasma membranes of boar sperm were applied to plastic wells coated with either of the two kind of oligosaccharide probes, several molecules bound to the wells and binding was inhibited by addition of corresponding oligosaccharides.4) A molecule (70 kDa) was shown to have an affinity to the N-acetyllactosamine structure by affinity chromatography. Partial amino acid sequences of N-terminal portion and proteolytic peptide fragments of the protein were determined. On the basis of the sequence, CDNA encoding the protein could be cloned and its nucleotide sequence was determined.These results will help us further analysis of fertilization on the molecular level.