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Identification of fucosylated glycoconjugates in Xenopus laevis testis by lectin histochemistry

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Abstract

Glycoconjugates play roles in many physiological and pathological processes. Previous works have shown important functions mediated by glycans in spermatogenesis, and the carbohydrate composition of testis has been studied by several approaches, including lectin-histochemical methods. However, the testis of Xenopus laevis, an animal model extensively employed in biochemical, cell and developmental research, has not yet been analysed. The aim of this work was to carry out a histochemical study of the fucose (Fuc)-containing glycoconjugates of Xenopus testis by means of lectins, combined with deglycosylation pretreatments. Four Fuc-binding lectins were used: orange peel (Aleuria aurantia) lectin (AAL), gorse seed (Ulex europaeus) agglutinin-I (UEA-I), fresh water eel (Anguilla anguilla) agglutinin (AAA), and asparagus pea (Lotus tetragonolobus) agglutinin (LTA), each recognizing different forms of fucosylated glycans. Labelling with UEA-I, which preferably binds Fucα(1,2) containing oligosaccharides, did not show any appreciable staining. LTA, specific for Fucα(1,3), and AAA, which binds Fucα(1,2), labelled spermatocytes and spermatids, but no labelling was seen when the histochemical procedure was carried out after either β-elimination (which removes O-linked oligosaccharides) or incubation with PNGase F (which removes N-linked oligosaccharides), suggesting that fucosylated glycans are of both N- and O-linked types. AAL, which has its highest affinity to Fucα(1,6), but also recognizes Fucα(1,2) and Fucα(1,3), labelled the whole testis, and the staining remained when the histochemical method was performed after either β-elimination or incubation with PNGase F. Labelling with AAL could be explained by the fact that this lectin could be binding to diverse fucosylated glycans in N- and O-glycans, and even in glycolipids. The importance of these glycans is discussed.

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Acknowledgments

This work was supported by grants of the UPV/EHU (1/UPV00075.310-E-14847/2002 and 1/UPV00077.310-E-15927/2004) and Fundación Séneca (Comunidad Autónoma de la Región de Murcia, grant number 04542/GERM/06). G.V. was a fellowship from the UPV/EHU. Mrs M. Portuondo and Mrs. C. Otamendi contributed to sample preparation. We thank Mrs M.J. Aldasoro for her support in the office work.

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  1. Departamento de Biología Celular e Histología, Facultad de Medicina y Odontología, Universidad del País Vasco/Euskal Herriko Unibertsitatea, Bº Sarriena s/n, 48940, Leioa (Vizcaya), Spain

    Galder Valbuena & Francisco José Sáez

  2. Department of Cell Biology and Histology, University of Murcia, Espinardo (Murcia), Spain

    Juan Francisco Madrid & Francisco Hernández

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  1. Galder Valbuena
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Correspondence to Francisco José Sáez.

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Valbuena, G., Madrid, J.F., Hernández, F. et al. Identification of fucosylated glycoconjugates in Xenopus laevis testis by lectin histochemistry. Histochem Cell Biol 134, 215–225 (2010). https://doi.org/10.1007/s00418-010-0722-x

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