Abstract
It is well known that cell surface glycans or glycocalyx play important roles in sperm motility, maturation and fertilization. A comprehensive profile of the sperm surface glycans will greatly facilitate both basic research (sperm glycobiology) and clinical studies, such as diagnostics of infertility. As a group of natural glycan binders, lectin is an ideal tool for cell surface glycan profiling. However, because of the lack of effective technology, only a few lectins have been tested for lectin-sperm binding profiles. To address this challenge, we have developed a procedure for high-throughput probing of mammalian sperm with 91 lectins on lectin microarrays. Normal sperm from human, boar, bull, goat and rabbit were collected and analyzed on the lectin microarrays. Positive bindings of a set of ~50 lectins were observed for all the sperm of 5 species, which indicated a wide range of glycans are on the surface of mammalian sperm. Species specific lectin bindings were also observed. Clustering analysis revealed that the distances of the five species according to the lectin binding profiles are consistent with that of the genome sequence based phylogenetic tree except for rabbit. The procedure that we established in this study could be generally applicable for sperm from other species or defect sperm from the same species. We believe the lectin binding profiles of the mammalian sperm that we established in this study are valuable for both basic research and clinical studies.
Highlights
The membrane surface of sperm is coated with a thick layer of glycans, i.e., the sperm glycocalyx, which is the first interface between sperm and the environment [1]
To obtain a comprehensive profile of the accessible glycans on mammalian sperm surface, we have developed a procedure for high-throughput probing of mammalian sperm on a lectin microarray with 91 lectins
In addition to traditional steps for probing intact cells on lectin microarray, i.e., labeling, binding, washing and readout, a fixation step is included for sperm analysis
Summary
The membrane surface of sperm is coated with a thick layer of glycans, i.e., the sperm glycocalyx, which is the first interface between sperm and the environment [1]. Enzyme, biotin or fluorescent dye conjugated lectin was individually applied for staining and the binding results were readout by microscope or flow-cytometry [16], the basic methodologies for lectin based analysis are immunocytochemistry, immunohistochemistry and flow cytometry, which are labor-intensive, time-consuming, and often require cumbersome sample pre-treatment. These methodologies are usually carried out in a one by one fashion, which is not suitable for global and highthroughput analysis
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