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Vol. 13, Issue 8, 2810-2825, August 2002

Structure of the Golgi and Distribution of Reporter Molecules at 20°C Reveals the Complexity of the Exit Compartments

Mark S. Ladinsky,^* Christine C. Wu, Shane McIntosh, J. Richard McIntosh,^* and Kathryn E. Howell^*§

 ^*Boulder Laboratory for 3-D Fine Structure, Department of Molecular, Cellular, and Developmental Biology, University of Colorado, Boulder, Colorado 80309; and  Department of Cellular and Structural Biology, University of Colorado Medical School, University of Colorado, Denver, Colorado 80262

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.

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^§ Corresponding author. E-mail address: Kathryn.Howell{at}UCHSC. edu.

Present address: Department of Cell Biology, Scripps Research Institute, La Jolla, CA 92037.

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Molecular Biology of the Cell
Vol. 13, 2810-2825, August 2002
Copyright © 2002 by The American Society for Cell Biology

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