Identification of fucosylated glycoconjugates in Xenopus laevis testis by lectin histochemistry.
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 Fucalpha(1,2) containing oligosaccharides, did not show any appreciable staining. LTA, specific for Fucalpha(1,3), and AAA, which binds Fucalpha(1,2), labelled spermatocytes and spermatids, but no labelling was seen when the histochemical procedure was carried out after either beta-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 Fucalpha(1,6), but also recognizes Fucalpha(1,2) and Fucalpha(1,3), labelled the whole testis, and the staining remained when the histochemical method was performed after either beta-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.
Valbuena
Galder
G
Departamento de Biología Celular e Histología, Facultad de Medicina y Odontología, Universidad del País Vasco/Euskal Herriko Unibertsitatea, 48940 Leioa (Vizcaya), Spain.
Madrid
Juan Francisco
JF
Hernández
Francisco
F
Sáez
Francisco José
FJ
D000818:Animals; D005643:Fucose / Q000032:analysis / Q000378:metabolism; D006001:Glycoconjugates / Q000032:analysis* / Q000378:metabolism; D006651:Histocytochemistry / Q000379:methods; D037102:Lectins; D008297:Male; D037121:Plant Lectins; D013087:Spermatids / Q000737:chemistry; D013090:Spermatocytes / Q000737:chemistry; D013091:Spermatogenesis; D013737:Testis / Q000737:chemistry*; D014982:Xenopus laevis
D006001:Glycoconjugates; D037102:Lectins; D037121:Plant Lectins; C027112:fucose-binding lectin; D005643:Fucose