Research Article

Identification of mannose moieties in N- and O-linked oligosaccharides of the primordial germ cells of Xenopus embryos

  1. Edurne Alonso1,
  2. Laura Gómez1,
  3. Juan Francisco Madrid2,
  4. Francisco José Sáez1,*

Article first published online: 23 MAY 2006

DOI: 10.1002/jemt.20318

Issue

Microscopy Research and Technique

Microscopy Research and Technique

Special Issue: “Nanomaterials Characterization Using Microscopy—Part II”

Volume 69, Issue 7, pages 595–599, July 2006

Additional Information

How to Cite

Alonso, E., Gómez, L., Madrid, J. F. and Sáez, F. J. (2006), Identification of mannose moieties in N- and O-linked oligosaccharides of the primordial germ cells of Xenopus embryos. Microsc. Res. Tech., 69: 595–599. doi: 10.1002/jemt.20318

Author Information

  1. 1

    Departamento de Biología Celular e Histología, Facultad de Ciencia y Tecnología, Universidad del País Vasco/Euskal Herriko Unibertsitatea, B° Sarriena s/n, E-48940 Leioa (Vizcaya), Spain

  2. 2

    Departamento de Biología Celular, Facultad de Medicina, Universidad de Murcia, E-30100 Espinardo (Murcia), Spain

*Universidad del País Vasco, Departamento de Biología Celular e Histología, B° Sarriena s/n, E-48940 Leioa (Vizcaya), Spain

Publication History

  1. Issue published online: 26 JUN 2006
  2. Article first published online: 23 MAY 2006
  3. Manuscript Accepted: 31 JAN 2006
  4. Manuscript Received: 28 SEP 2005

Funded by

  • Universidad del País Vasco/Euskal Herriko Unibertsitatea. Grant Numbers: 075.310-EA137/97, 075.327-G10/99
  • Spanish Ministerio de Educación y Ciencia

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Keywords:

  • glycans;
  • glycome;
  • lectins;
  • histochemistry;
  • deglycosylation;
  • cell migration;
  • glycoconjugates

Abstract

The presence of mannose (Man) in the glycoconjugates of primordial germ cells (PGCs) of Xenopus embryos was elucidated by lectin histochemistry with Concanavalin A (Con A) and snowdrop (Galanthus nivalis) bulb lectin (GNA), in combination with deglycosylative pretreatments: β-elimination, which removes O-linked oligosaccharides, and incubation with Peptide N glycosidase F (PNGase F), which removes N-linked glycan chains. In addition, histochemistry with Con A, which binds to Man and glucose (Glc), was also performed after glucose-oxidase incubation, which converts Glc into gluconic acid, and GNA was carried out after acid hydrolysis, which removes terminal sialic acid (NeuAc) moieties. PGCs were analyzed during their migration over the mesentery until the genital ridge, and after colonization of this gonad anlage. The results showed that for both lectins: (1) the PGCs and other surrounding tissue showed a similar binding pattern, and (2) the staining in the PGCs was similar in the developmental stages studied. Labeling with Con A was due to Man, and not to Glc, as shown after incubation with glucose-oxidase, and it was assumed that Man was in N-linked oligosaccharides. However, GNA labeling was mainly due to O-linked oligosaccharides, because the pretreatment of β-elimination turned cells negative. Moreover, acid hydrolysis pretreatment gave rise to a stronger GNA-staining, suggesting that either Man was also in subterminal position to NeuAc or some Man-containing glycans were unmasked after removal of NeuAc from other oligosaccharide chains. Microsc. Res. Tech., 2006. © 2006 Wiley-Liss, Inc.

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