Wheat germ agglutinin stains dispersed post-golgi vesicles after treatment with the cytokinesis inhibitor psychosine.
The galactosylsphingosine psychosine (Psy) is one of the sphingolipids and induce the formation of multinuclear cells in several cell lines by inhibiting cytokinesis. In the present report, we show that intracellular organelles, including wheat germ agglutinin (WGA)-positive vesicles and early endosomes, are selectively dispersed by Psy. WGA is a conventional Golgi marker and WGA-positive vesicles appeared to co-localize with the Golgi apparatus in untreated cells. Psy treatment induced the dispersal of WGA-positive vesicles without affecting the structure of the Golgi apparatus, resulting in discrimination of WGA-positive vesicles from the Golgi apparatus. In sharp contrast to this effect of Psy, WGA-positive vesicles were not affected by brefeldin A treatment, which induced the disappearance of the Golgi apparatus. Immunostaining with anti-TGN46 antibodies revealed that a large portion of the WGA-positive vesicles were derived from the trans-Golgi network. Notably, the dispersed WGA-positive vesicles did not stain with anti-syntaxin 6, another marker of the trans-Golgi network. During cytokinesis, WGA-positive vesicles in the cytoplasm decreased, and WGA staining accumulated at the cleavage furrow, which was apparently inhibited by the presence of Psy. These data suggest that the transport of WGA-positive vesicles to the cleavage furrow is associated with the progression of cytokinesis.
Kanazawa
Takayuki
T
Laboratory of Membrane Biochemistry and Biophysics, Graduate School of Biostudies, Kyoto University, Kyoto, Japan.
Takematsu
Hiromu
H
Yamamoto
Akitsugu
A
Yamamoto
Harumi
H
Kozutsumi
Yasunori
Y
D000818:Animals; D000906:Antibodies; D020126:Brefeldin A / Q000494:pharmacology; D019556:COS Cells; D002522:Chlorocebus aethiops; D048749:Cytokinesis / Q000187:drug effects*; D022162:Cytoplasmic Vesicles / Q000648:ultrastructure; D004705:Endocytosis / Q000187:drug effects; D011992:Endosomes / Q000187:drug effects / Q000648:ultrastructure; D006056:Golgi Apparatus / Q000187:drug effects / Q000648:ultrastructure*; D007158:Immunologic Techniques; D008562:Membrane Glycoproteins / Q000276:immunology; D008854:Microscopy, Electron; D008870:Microtubules / Q000187:drug effects / Q000648:ultrastructure; D015388:Organelles / Q000187:drug effects / Q000648:ultrastructure*; D011609:Psychosine / Q000494:pharmacology*; D013194:Staining and Labeling / Q000379:methods; D014909:Wheat Germ Agglutinins; D021601:trans-Golgi Network / Q000648:ultrastructure
D000906:Antibodies; D008562:Membrane Glycoproteins; C072237:TGOLN2 protein, human; D014909:Wheat Germ Agglutinins; D020126:Brefeldin A; D011609:Psychosine