Nonuniform microtubular polarity established by CHO1/MKLP1 motor protein is necessary for process formation of podocytes

Podocytes are unique cells that are decisively involved in glomerular filtr ation. They are equipped with a complex process system consisting of major processes and foot processes whose function is insufficiently understood (M undel, P., and W. Kriz. 1995. Anat. Embryol. 192:385-397). The major proces ses of podocytes contain a microtubular cytoskeleton. Taking advantage of a recently established cell culture system for podocytes with preserved abil ity to form processes (Mundel, P,, J. Reiser, A. Zuniga Mejia Borja, H. Pav enstadt, G.R. Davidson, W. Kriz, and R, Zeller, 1997b. Exp. Cell Res. 36:24 8-258), we studied the functional significance of the microtubular system i n major processes. The following data were obtained: (a) Microtubules (MTs) in podocytes show a nonuniform polarity as revealed by hook-decoration. (b ) CHO1/ MKLP1, a kinesin-like motor protein, is associated with MTs in podo cytes. (c) Treatment of differentiating podocytes with CHO1/MKLP1 antisense oligonucleotides abolished the formation of processes and the nonuniform p olarity of MTs. (d) During the recovery from taxol treatment, taxol-stabili zed (nocodazole-resistant) MT fragments were distributed in the cell periph ery along newly assembled nocodazole-sensitive MTs, ii similar distribution pattern of CHO1/MKLP1 was found under these circumstances, indicating its association with MTs. (e) In the recovery phase after complete depolymeriza tion, MTs reassembled exclusively at centrosomes. Taken together, these fin dings lead to the conclusion that the nonuniform MT polarity in podocytes e stablished by CHO1/MKLP1 is necessary for process formation.