A new histochemical approach for studying sperm cell surfaces

Ngo, Lylla; Barajas, Marcella; Weerasinghe, Gayani; Zem, Gregory; Oppenheimer, Steven B.

doi:10.1078/0065-1281-00689

Cited by 4 publications

(3 citation statements)

References 7 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…However, as we note above, it seems likely that binding by all four lectins was not completely specific to their target monosaccharides (see also Fallis et al 46 for similar finding in other mussel species). Such findings are perhaps not surprising given that the biological ligands of the lectins are more complex than single monosaccharides and lectins have significantly higher binding affinity on such complex carbohydrate structures 39 46 47 48 49 . Thus, the demonstration of truly non-specific binding would require the use of more complex inhibiting carbohydrates.…”

Section: Discussionmentioning

confidence: 98%

“…This conclusion is also supported by the fact that both the overall binding affinity (fluorescence intensity) and monosaccharide non-specificity was found to be highest for WGA, a lectin which is known to have especially broad carbohydrate specificity 50 . Furthermore, the high salt concentration of the seawater can prevent binding on structurally simple monosaccharides (such as inhibiting hapten sugars), but may not severely impede binding to more complex sperm surface carbohydrates as these bindings are structurally stronger 47 48 . Taken together, although we cannot exclude the possibility of some non-specific binding, it is likely that a large majority of the observed lectin binding was specific to their target glycan structures.…”

Section: Discussionmentioning

confidence: 99%

See 1 more Smart Citation

Lectin staining and flow cytometry reveals female-induced sperm acrosome reaction and surface carbohydrate reorganization

Kekäläinen

Larma

Linden

et al. 2015

Sci Rep

View full text Add to dashboard Cite

All cells are covered by glycans, an individually unique layer of oligo- and polysaccharides that are critical moderators of self-recognition and other cellular-level interactions (e.g. fertilization). The functional similarity between these processes suggests that gamete surface glycans may also have an important, but currently overlooked, role in sexual selection. Here we develop a user-friendly methodological approach designed to facilitate future tests of this possibility. Our proposed method is based on flow cytometric quantification of female-induced sperm acrosome reaction and sperm surface glycan modifications in the Mediterranean mussel Mytilus galloprovincialis. In this species, as with many other taxa, eggs release water-soluble factors that attract conspecific sperm (chemoattraction) and promote potentially measurable changes in sperm behavior and physiology. We demonstrate that flow cytometry is able to identify sperm from other seawater particles as well as accurately measure both acrosome reaction and structural modifications in sperm glycans. This methodological approach can increase our understanding of chemically-moderated gamete-level interactions and individual-specific gamete recognition in Mytilus sp. and other taxa with similar, easily identifiable acrosome structure. Our approach is also likely to be applicable to several other species, since carbohydrate-mediated cellular-level interactions between gametes are universal among externally and internally fertilizing species.

show abstract

Section: Discussionmentioning

confidence: 98%

Section: Discussionmentioning

confidence: 99%

Lectin staining and flow cytometry reveals female-induced sperm acrosome reaction and surface carbohydrate reorganization

Kekäläinen

Larma

Linden

et al. 2015

Sci Rep

View full text Add to dashboard Cite

show abstract

“…Over the past decade we have used microbeads derivatized with over 100 different compounds to study cell surface properties (Ngo et al, 2003;Khurrum et al, 2003;Navarro et al, 2002;Salbilla et al, 1999;Latham and Oppenheimer, 1999;Heinrich et al, 2005;Welty et al 2005;Latham et al 1995;1995a). We are unaware of any studies that have defined the microbead system as we have done in the present paper.…”

Section: Discussionmentioning

confidence: 99%

Microbead analysis of cell binding to immobilized lectin: an alternative to microarrays in the development of carbohydrate drugs and diagnostic tests

et al. 2006

Self Cite

View full text Add to dashboard Cite

SummaryMicroarray technology is currently used in the development of carbohydrate drugs and diagnostic tests. Here we model an inexpensive alternative to microarrays using derivatized microbeads. In this model we examine the binding of mannose-rich yeast to microbeads derivatized with concanavalin A (Con A), a mannose-binding lectin, in the presence of 30 different sugars and 9 different pH conditions. We developed a listing of effective saccharide inhibitors of immobilized Con A based on 3901 replicates. We suggest that this is the most extensive saccharide inhibitor list ever developed for this lectin and it may be useful to use this listing to replace the less extensive lists that have been in the literature for decades. Information is also provided on pH effects on immobilized Con A binding based on 918 trials. Two assays to study binding, one which qualitatively scores more or less binding than control in thousands of replicate samples, and another that quantitatively evaluates binding by counting the number of cells bound to each bead, are also modeled here. We know of no previous studies that provide such as extensive information on saccharide inhibition and pH effects on the binding of immobilized Con A. We suggest that this microbead approach, using beads derivatized with lectins or sugars, and the two simple assays presented here, can in some cases, substitute for more expensive microarray technology in the development of carbohydrate drugs and diagnostic tests. If, for example, our model Saccharomyces cerevisiae was a pathogen, these studies show that it binds via cell surface mannose residues and drugs to prevent binding could be developed using the inhibitors of binding identified here. The beads could be also used in the development of diagnostic tests that identify the presence of the organism in blood samples, etc. in much the same way as microarray technology is being used today.

show abstract