Effect of Caffeine and Reduced Temperature (20°C) on the Organization of the Pre-Golgi and the Golgi Stack Membranes

• Published in: The Journal of cell biology 1993-03, Vol.120 (6), p.1321-1335
• Main Authors: Jäntti, Jussi, Kuismanen, Esa
• Format: Article
• Language: English
• Subjects: Animals; Anti-Bacterial Agents - pharmacology; Antibodies; Biological transport; Biological Transport - drug effects; Brefeldin A; Caffeine; Caffeine - pharmacology; Cell Line; Cells; Cellular biology; Cellular immunity; Cricetinae; Cyclopentanes - pharmacology; Endoplasmic Reticulum - drug effects; Endoplasmic Reticulum - metabolism; Endoplasmic Reticulum - ultrastructure; Fluorescent antibody techniques; Glycoproteins; Golgi apparatus; Golgi Apparatus - drug effects; Golgi Apparatus - metabolism; Golgi Apparatus - ultrastructure; Intracellular Membranes - drug effects; Intracellular Membranes - metabolism; Intracellular Membranes - ultrastructure; Kidney; Kidney cells; Microscopy, Electron; P branes; String theory; Temperature
• ISSN: 0021-9525; 1540-8140
• DOI: 10.1083/jcb.120.6.1321

In the present study we have dissected the transport pathways between the ER and the Golgi complex using a recently introduced inhibition of transport by caffeine at 20°C. Recovery of the Golgi complex from brefeldin A (BFA) treatment was inhibited by caffeine at reduced temperature (20°C) suggesting that caffeine inhibits the membrane traffic between the ER and the Golgi complex. Caffeine at 20°C did not inhibit the BFA-induced retrograde movement of the Golgi membranes. Further, incubation of the cells in 10 mM caffeine at 20°C had profound effects on the distribution and the organization of the pre-Golgi and the Golgi stack membranes. Caffeine treatment at 20°C resulted in a selective and reversible translocation of the pre- and cis-Golgi marker protein (p58) to the periphery of the cell. This caffeine-induced effect on the Golgi complex was different from that induced by BFA, since mannosidase II, a Golgi stack marker, remained perinuclearly located and the Golgi stack coat protein, β-COP, was not detached from Golgi membranes in the presence of 10 mM caffeine at 20°C. Electron microscopic analysis showed that, in the presence of caffeine at 20°C, the morphology of the Golgi stack was altered and accumulation of numerous small vesicles in the Golgi region was observed. The results in the present study suggest that caffeine at reduced temperature (20°C) reveals a functional interface between the pre-Golgi and the Golgi stack.