Cold and metabolic inhibitor effects on cytoplasmic microtubules and the Golgi complex in cultured rat epiphyseal chondrocytes

Abstract

Previous work has shown that exposure of cultured chondrocytes to colchicine leads to disappearance of microtubules and dispersion of the dictyosomes of the Golgi complex throughout the cytoplasm. Here, the effects of cold and metabolic inhibitors on cultured chondrocytes have been investigated in order to characterize further the relationship between these organelle systems. After incubation of cells for 24h at 4 degrees C most, but not all microtubules disappeared, indicating the existence of cold-resistant microtubules. Dictyosomes remained united in one area, until transfer of cultures to 37 degrees C, when they dispersed throughout the cytoplasm in about one-third of the cells. In cells exposed simultaneously to cold and colchicine, microtubules disappeared completely, but spreading of dictyosomes occurred only in some cells and became generalized first upon warming. Application of the metabolic inhibitors sodium azide or sodium fluoride (10(-2) M) or 2-deoxyglucose (5 X 10(-2) M) together with sodium cyanide (10(-2) M) inhibited microtubule removal by colchicine. Consequently, spreading of the Golgi complex was prevented. These findings support the concept of an important role of microtubules in the organization of the Golgi complex. Moreover, depolymerization of microtubules by colchicine appears to be an energy dependent process.