LOCOMOTION OF CV-1 CYTOPLASTS CONTAINING OR LACKING CENTROSOME
A. V. Burakov
The A. N. Belozersky Institute for Physico-Chemical Biology, Moscow State University;
e-mail: antburakov@cmail.ru
The movement of cultured cells along the substratum is a convenient model for studying cell movement in the
organism, occurring during embryogenesis, angiogenesis, metastasis, wound closure, etc. The moving cells must control
their pseudopodial activity along the perimeter, regulate the attachment and reattachment to the substratum, and
pull their body following pseudopodium during their movement along the substratum. As proven by numerous
investigations, these processes directly depend on the actomyosin system of cells. The role of microtubules as
components of cytoskeleton in cell locomotion still remains obscure. The role of microtubules in cell movement is
commonly investigated using microtubule-destructive drugs. Therefore in the final results the accessory drug effect
on, for example, signal cascades cannot be excluded. Another mode of action on the microtubule dynamics is centrosome
removal from the cells, which is easily realized by their removal together with the nucleus. It has been shown that
in cytoplasts of centrosome containing fibroblasts, dynamic instability of microtubules remains. Unlike, in
non-centriolar cytoplasts tread milling is observed. Some literature evidence suggests that cytoplasts of cultured
cells move along the substratum not worse that intact cells do. In this study cytoplasts with and without centrosome
were obtained and identified, and parameters of movement along the substratum (speed, direction) were registered for
both these two populations of cytoplasts, and for control intact cells and cells involved in the experiment. The
model of experimental wound of monolayer was used, because it guaranteed cell synchronization in respect to
movement direction and speed. Centrosome-containing CV-1 cytoplasts displayed radial microtubule array, and
centrosome-lacking cytoplasts exhibited chaotic distribution of microtubules, which is characteristic of microtubule
tread milling. In addition, both kinds of cytoplasts appeared to move along the substratum much slower than the whole
cells. No difference was found is speed and keeping direction between centriolar and non-centriolar cytoplasts.
Key words: cytoskeleton, microtubules, locomotion, centrosome, cytoplasts
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