Interaction of microtubules with peroxisomes. Tubular and spherical peroxisomes in HepG2 cells and their alterations induced by microtubule-active drugs.
We have studied the interaction of microtubules with peroxisomes and the influence of changes in the microtubular network on the peroxisomal compartment. From the several cell lines analyzed for this purpose, HepG2 cells proved to be the best candidate exhibiting both a well-developed cytoskeleton and a peroxisomal compartment with great plasticity. Three distinct types of peroxisomes: small spherical (0.1-0.3 micron), rod-shaped (0.5 micron) and elongated tubular (up to 5 microns) ones were identified in this cell line. A shift of the elongated tubular forms to spherical particles was noted by increasing the density of cells in culture, whereas no correlation between the distinct peroxisomal forms and the cellular proliferation could be observed. At time points when the elongated tubular peroxisomes were disappearing, many spherical peroxisomes arranged like 'chains of beads on a string' were observed, suggesting that the fission of elongated tubular forms may give rise to newly developing spherical peroxisomes. A clear association of spherical peroxisomes with microtubules was visualized by double immunofluorescence in combination with confocal laser scanning microscopy (CLSM). Treatment with a variety of microtubule-depolymerizing drugs (colcemid, nocodazole, vinblastine) induced a significant increase in the frequency of tubular peroxisomes and led to the formation of peroxisomal clusters. These effects were reversible since already 1 to 2 h after removal of the drugs from the culture medium, a uniform distribution of spherical peroxisomes was reestablished. Taxol, a microtubule-stabilizing drug, on the other hand exerted no significant effects on the peroxisomal compartment. The direct interaction of microtubules with peroxisomes in vitro was demonstrated using highly purified rat liver peroxisomes and taxol-stabilized microtubules from bovine or pig brain. The binding of peroxisomes to microtubules was visualized by video-enhanced contrast microscopy (VECM) and was abolished by pretreatment of peroxisomes with 100 mM KCl ('stripping'), proteinase K or trypsin. Incubation with cytosol restored the binding capacity of KCl-treated peroxisomes, but did not complement the protease treatment. The data presented provide for the first time evidence for a direct interaction of microtubules with the peroxisomal compartment indicating that this cytoskeletal system plays an important role in the morphogenesis and intracellular distribution of peroxisomes.
Schrader
M
M
Institute for Anatomy and Cell Biology II, University of Heidelberg, Germany.
Burkhardt
J K
JK
Baumgart
E
E
Lüers
G
G
Spring
H
H
Völkl
A
A
Fahimi
H D
HD
D008841:Actin Cytoskeleton / Q000378:metabolism; D000818:Animals; D016466:CHO Cells; D006224:Cricetinae; D003571:Cytochalasin B / Q000494:pharmacology; D015638:Cytochalasin D / Q000494:pharmacology; D003703:Demecolcine / Q000494:pharmacology; D006571:Heterocyclic Compounds / Q000494:pharmacology*; D006801:Humans; D008830:Microbodies / Q000378:metabolism* / Q000648:ultrastructure; D018613:Microscopy, Confocal; D008870:Microtubules / Q000187:drug effects / Q000378:metabolism* / Q000502:physiology; D015739:Nocodazole / Q000494:pharmacology; D017239:Paclitaxel / Q000494:pharmacology; D051381:Rats; D014407:Tumor Cells, Cultured; D014747:Vinblastine / Q000494:pharmacology
D006571:Heterocyclic Compounds; D015638:Cytochalasin D; D003571:Cytochalasin B; D014747:Vinblastine; D017239:Paclitaxel; D015739:Nocodazole; D003703:Demecolcine