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CIL:8059*  Cite 

Brefeldin A (BFA) is a fungal metabolite that reversibly blocks protein transport from the endoplasmic reticulum (ER) to the Golgi apparatus. This effect is accompanied by a distinctive morphological change, the reversible collapse of the Golgi apparatus. The disassembly of the Golgi apparatus is accompanied by its dramatic tubulation, forming a network extending throughout the cytoplasm. While the content of the Golgi apparatus is redistributed to the ER, the content of the trans-Golgi network (TGN) collapses in the vicinity of the MTOC. This video shows the effect of BFA in an NRK cell stably expressing GFP-GMx33a, a protein associated to the TGN. NRK cells stably expressing GFP-GMx33a and plated on 35-mm glass-bottom culture dishes (MatTek), were treated with 5 microgram/ml BFA and maintained in phenol red-free buffered medium at 37C by using an ASI 400 Air Stream stage Incubator (Nevtek, Burnsville, VA). After 10 minutes in the presence of BFA time-lapse fluorescence images were acquired with an Ultraview Confocal Scanner (Perkin Elmer-Cetus, Norwalk, CT) in a Nikon Eclipse TE2000-S inverted microscope (Nikon, Melville, NY) equipped with a PlanApo 100x oil immersion objective (NA 1.40; Nikon), and a 12-bit charge-coupled device (CCD) camera, ORCA (Hamamatsu, Bridgewater, NJ). Image capture and data acquisition were performed using Ultraview LCI software (Perkin Elmer-Cetus). Images were acquired in binning 2 x 2 modes at approximately 0.25-s intervals, during approximately 175 s. Sequence images were exported as single TIFF files, and processed with ImageJ 1.36b (Rasband, 1997, 2007; Lippincott-Schwartz J, Donaldson JG, Schweizer A, Berger EG, Hauri HP, Yuan LC, Klausner RD. Microtubule-dependent retrograde transport of proteins into the ER in the presence of brefeldin A suggests an ER recycling pathway. Cell [serial online]. 1990;60:821-36. Lippincott-Schwartz J, Yuan L, Tipper C, Amherdt M, Orci L, Klausner RD. Brefeldin A's effects on endosomes, lysosomes, and the TGN suggest a general mechanism for regulating organelle structure and membrane traffic. Cell [serial online]. 1991;67:601-16. Snyder CM, Mardones GA, Ladinsky MS, Howell KE. GMx33 associates with the trans-Golgi matrix in a dynamic manner and sorts within tubules exiting the Golgi. Mol Biol Cell [serial online]. 2006;17:511-24.

Biological Sources
NCBI Organism Classification
Rattus norvegicus
Cell Type
permanent cell line cell
Cell Line
Cellular Component
Golgi apparatus part
Biological Context
Biological Process
Mardones, Gonzalo A.
Digital Object Identifier (DOI)
Archival Resource Key (ARK)
Grouping This image is part of a group.
Sample Preparation
living tissue
Relation To Intact Cell
whole mounted tissue
Spatial Axis Image Size Pixel Size
X 328px 150nm
Y 392px 150nm
Time 175 seconds