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Volume 16, Number 14, Issue of July 15, 1996 pp. 4370-4375
Copyright ©1996 Society for Neuroscience

Expression of a Kinesin-Related Motor Protein Induces Sf9 Cells to Form Dendrite-Like Processes with Nonuniform Microtubule Polarity Orientation

Received March 1, 1996; revised April 23, 1996; accepted April 29, 1996.

David J. Sharp¹, Ryoko Kuriyama², and Peter W. Baas¹

¹ Department of Anatomy and Program in Neuroscience, The University of Wisconsin Medical School, Madison, Wisconsin 53706, and ² Department of Cell Biology and Neuroanatomy, The University of Minnesota Medical School, Minneapolis, Minnesota 55455

The microtubules (MTs) within neuronal processes are highly organized with regard to their polarity and yet are not attached to any detectable nucleating structure. Axonal MTs are uniformly oriented with their plus ends distal to the cell body, whereas dendritic MTs are of both orientations. Here, we sought to test the capacity of motor-driven MT transport to organize distinct MT patterns during process outgrowth. We focused on CHO1/MKLP1, a kinesin-related protein present in the midzonal region of the mitotic spindle where MTs of opposite orientation overlap. Insect ovarian Sf9 cells induced to express the N-terminal portion of the molecule form MT-rich processes with a morphology similar to that of neuronal dendrites (). Nascent processes contain uniformly plus-end-distal MTs, but these are joined by minus-end-distal MTs as the processes continue to develop. Thus, this CHO1/MKLP1 fragment establishes a nonuniform MT polarity pattern and does so by a similar sequence of events as occurs with the dendrite, the antecedent of which is a short process with a uniform MT polarity orientation. Two lines of evidence suggest that these results are elicited by motor-driven MT transport. First, there is a depletion of MTs from the cell body during process outgrowth. Second, the same polarity pattern is obtained when net MT assembly is suppressed pharmacologically during process formation. Collectively, these findings provide precedent for the idea that motor-driven transport can organize MTs into distinct patterns of polarity orientation during process outgrowth.

Key words: microtubule; kinesin; dendrite; neuron; mitosis; spindle

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