Real-time observation of the disassembly of stable neuritic microtubules induced by laser transection: possible mechanisms of microtubule stabilization in neurites.

Abstract

By dissolving the membrane with detergent perfusion, we have shown that the established neurites of dorsal root ganglion cells cultured for more than 5 days contained microtubules which persisted outside the cell for a few minutes to more than 1h [Tashiro et al., 1997: J. Neurosci. Res. 50:81-93]. To investigate their stabilization mechanism, we transected the exposed microtubules by laser microbeam irradiation and observed their length changes with video-enhanced differential interference contrast microscopy. Microtubule fragments started to shorten on both sides of the transection site. more rapidly from the newly generated plus ends than from the minus ends. The maximal rate as well as the pattern of shortening correlated with the time of transection; microtubules transected later than 30 min after membrane removal shortened at rates less than 20 microm/min and typically with intermittent pauses, while the more labile microtubules included in the earlier transections shortened continuously at higher rates. Microtubules in neurites were thus stabilized by 1) stopping disassembly at local sites including the plus ends, and 2) slowing disassembly along the length. Transection also suggested the presence of specialized points along microtubules which are involved in anchoring microtubules to the substratum. Cell Motil. Cytoskeleton 42:87-100, 1999.