Ultrastructural localization of polysaccharides and N-acetyl-D-galactosamine in the secretory pathway of green algae (Demidiaceae)
1998
Brosch-Salomon, S. | Hoftberger, M. | Holzinger, A. | Lutz-Meindl, U.
In order to characterize the secretory pathway leading to multipolar tip growth in green algae of the family Desmidiaceae, different general polysaccharide stainings, such as the periodic acid-silver hexamine method and the periodic acid-silver proteinate method as well as different lectins specific to defined sugar residues have been employed. General polysaccharide stainings label different kinds of secretory vesicles starting from the onset of vesicle production up to their delivery into the primary cell wall, however, the discrimination of Golgi products is possible using lectins. Both gold-labelled lectins from Helix pomatia and from Glycine max with affinity to N-acetyl-D-galactosamine only produce labelling of primary wall material containing 'dark vesicles' on ultrathin sections of high-pressure frozen and freeze-substituted Micrasterias cells, whereas other vesicle types remain unstained. 'Dark vesicles' are labelled when still attached to trans-Golgi compartments, when distributed throughout the cytoplasm or when fusing with the plasma membrane with the same staining intensity which indicates that the sugars detected by the methods used are present from the onset of visible vesicle production. Gold-labelling of N-acetyl-D-galactosamine is also observed in the primary cell wall. In control experiments the staining vanishes when ultrathin sections are pre-incubated with N-acetyl-D-galactosamine. Various other lectins with affinity to different sugar residues than N-acetyl-D-galactosamine do not produce any staining of the cell wall nor of any kind of secretory vesicles. As N-acetyl-D-galactosamine is usually not present in N-linked polysaccharides the results point towards the presence of 0-linked-glycoproteins in the primary cell wall of desmids.
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