Journal of Andrology
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Published-Ahead-of-Print November 15, 2006, DOI:10.2164/jandrol.106.001073

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Characterizing the Glycocalyx of Poultry Spermatozoa: I. Identification and Distribution of Carbohydrate Residues using Flow Cytometry and Epifluorescence Microscopy

Jesus Pelaez and Julie A Long *

* To whom correspondence should be addressed. E-mail: jlong{at}anri.barc.usda.gov.

The aim of the present work was to use a battery of lectins to (1) delineate the carbohydrate content of sperm glycocalyx in the turkey and chicken using flow cytometry analysis and (2) evaluate the distribution of existing sugars over the sperm plasma membrane surface with epifluorescent microscopy. Carbohydrate groups (corresponding lectins) that were investigated included galactose (GS-I, Jacalin, RCA-I, PNA), glucose and/or mannose (Con A, PSA, GNA), N-acetyl-glucosamine (GS-II, s-WGA, STA), N-acetyl-galactosamine (SBA, WFA), fucose (Lotus, UEA-I), sialic acid (LFA, LPA) and N-acetyl-lactosamine (ECA). Sperm were assessed before and after treatment with neuraminidase to remove sialic acid. Mean Fluorescence Intensity (MnFI) was used as indicator of lectin binding for flow cytometry analysis. Non-treated sperm from both species showed high MnFI when incubated with RCA-I, Con A, LFA and LPA, as did chicken sperm incubated with s-WGA. Neuraminidase treatment increased the MnFI for most lectins except LFA and LPA, as expected. Differences in MnFI between species included higher values for s-WGA and ECA in chicken sperm and for WFA in turkey sperm. Microscopy revealed segregation of some sugar residues into membrane-specific domains; however, the two staining techniques (cell suspension versus fixed preparation) differed in identifying lectin binding patterns, with fixed preparations yielding a high degree of non-specific binding. We conclude that (1) the glycocalyx of turkey and chicken sperm contains a diversity of carbohydrate groups, (2) these residues are extensively masked by sialic acid, (3) the glycocalyx composition is species-specific, and (4) some glycoconjugates appear to be segregated into membrane-specific domains. Characterization of the poultry sperm glycocalyx is the first step in identifying the physiological impact of semen storage on sperm fucntion.


Key words: Semen • chicken • glycoconjugate • lectin • turkey




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J. Pelaez and J. A. Long
Characterizing the Glycocalyx of Poultry Spermatozoa: II. In Vitro Storage of Turkey Semen and Mobility Phenotype Affects the Carbohydrate Component of Sperm Membrane Glycoconjugates
J Androl, July 1, 2008; 29(4): 431 - 439.
[Abstract] [Full Text] [PDF]


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