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Vol. 13, Issue 8, 2810-2825, August 2002
*Boulder Laboratory for 3-D Fine Structure, Department of
Molecular, Cellular, and Developmental Biology, University of Colorado,
Boulder, Colorado 80309; and Incubating cells at 20°C blocks transport out of the Golgi
complex and amplifies the exit compartments. We have used the
20°C block, followed by EM tomography and serial section
reconstruction, to study the structure of Golgi exit sites in NRK
cells. The dominant feature of Golgi structure in temperature-blocked
cells is the presence of large bulging domains on the three trans-most
cisternae. These domains extend laterally from the stack and are
continuous with "cisternal" domains that maintain normal thickness
and alignment with the other stacked Golgi cisternae. The bulging
domains do not resemble the perpendicularly extending tubules
associated with the trans-cisternae of control cells. Such tubules are
completely absent in temperature-blocked cells. The three cisternae
with bulging domains can be identified as trans by their association with specialized ER and the presence of clathrin-coated buds on the
trans-most cisterna only. Immunogold labeling and
immunoblots show a significant degradation of a medial- and
a trans-Golgi marker with no evidence for their redistribution within
the Golgi or to other organelles. These data suggest that exit from the Golgi occurs directly from three trans-cisternae and that specialized ER plays a significant role in trans-Golgi function.
Department of Cellular and
Structural Biology, University of Colorado Medical School, University
of Colorado, Denver, Colorado 80262
Present address: Department of Cell Biology,
Scripps Research Institute, La Jolla, CA 92037.
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