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* Department of Biology, University of California at San Diego, La Jolla, California 92093-0322; and Peroxisomes in living CV1 cells were visualized by targeting the green fluorescent protein (GFP)
to this subcellular compartment through the addition of
a COOH-terminal peroxisomal targeting signal 1 (GFP-PTS1). The organelle dynamics were examined
and analyzed using time-lapse confocal laser scanning
microscopy. Two types of movement could be distinguished: a relatively slow, random, vibration-like movement displayed by the majority (~95%) of the peroxisomes, and a saltatory, fast directional movement displayed by a small subset (~5%) of the peroxisomes.
In the latter instance, peak velocities up to 0.75 µm/s
and sustained directional velocities up to 0.45 µm/s
over 11.5 µm were recorded. Only the directional type
of motion appeared to be energy dependent, whereas the vibrational movement continued even after the
cells were depleted of energy. Treatment of cells, transiently expressing GFP-PTS1, with microtubule-destabilizing agents such as nocodazole, vinblastine, and demecolcine clearly altered peroxisome morphology and
subcellular distribution and blocked the directional
movement. In contrast, the microtubule-stabilizing
compound paclitaxel, or the microfilament-destabilizing drugs cytochalasin B or D, did not exert these effects. High resolution confocal analysis of cells expressing GFP-PTS1 and stained with anti-tubulin antibodies
revealed that many peroxisomes were associated with
microtubules. The GFP-PTS1-labeled peroxisomes
were found to distribute themselves in a stochastic,
rather than ordered, manner to daughter cells at the
time of mitosis.
National Center for
Microscopy and Imaging Research, Department of Neurosciences, University of California at San Diego, La Jolla, California
92093-0608
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