Effects of brefeldin A on autophagy in cultured rat fibroblasts
Purhonen, P.; Pursiainen, K.; Reunanen, H.
European Journal of Cell Biology 74(1): 63-67
1997
ISSN/ISBN: 0171-9335 PMID: 9309391 Document Number: 482072
Effects of brefeldin A on cellular autophagy were studied in cultured rat fibroblasts. Brefeldin A inhibits the activation and membrane-binding properties of most ADP-ribosylation factors and causes the redistribution of Golgi proteins into the endoplasmic reticulum. Immunofluorescence and enzyme cytochemical methods revealed the disappearance of the Golgi apparatus and trans-Golgi network during the brefeldin A incubation. The volume fractions of autophagic vacuoles increased about threefold in cells treated with brefeldin A for 4 h and about six-fold in serum-deprived cells as compared with controls. When cells were first treated with brefeldin A for 1 h and were then deprived of serum and treated with brefeldin A for 3 h, the volume fraction of autophagic vacuoles increased about 4.5-fold as compared with untreated cells. The results showed that brefeldin A is unable to prevent serum deprivation-induced accumulation of autophagic vacuoles and that brefeldin A even when acting alone increases the volume fraction of autophagic vacuoles. It was concluded that an intact Golgi apparatus and trans-Golgi network are not essential for the formation of autophagic vacuoles. It seems also probable that ADP-ribosylation factors are not needed when vacuoles are formed.