Microtubule dynamics in serum-starved and serum-stimulated Swiss 3T3 mouse fibroblasts: implications for the relationship between serum-induced contractility and microtubules.
It has been established that cell contractility can be stimulated with low or depolymerizing doses of microtubule (MT) poisons. In addition, low doses of nocodazole and vinblastine have recently been shown to decrease MT dynamics in vivo. In this study, investigated whether there is a direct, or reciprocal feedback-type relationship between contractility and microtubule dynamics, by examining MT dynamic behavior in live cells under conditions where contractility is known to be altered. Quiescent, serum-starved Swiss 3T3 mouse fibroblasts have been shown to be weakened in their contractility; serum stimulation increases cell contractility and causes the formation of stress fibers and adhesion plaques. Growing (control), quiescent (Go), and serum-stimulated cells were injected with rhodamine-tubulin, and MT dynamics were determined by analysis of MT length changes obtained from digitized images of the extreme periphery of the cells, where the MT ends were readily apparent. The MTs in quiescent cells were less dynamic than those in control cells: the growth and shortening rates were reduced by 30% and 45%, respectively. Dynamicity decreased by 47%, and the MTs spent more time in pause. After serum stimulation, MT growth rate, dynamicity, and time spent in pause returned to control cell levels. Although the shortening rate increased by 28%, it remained significantly lower than in control cells. In this system, the serum-induced increase in contractility was accompanied by an increase in MT dynamics. However, increased contractility stimulated with low doses of MT poisons is known to be accompanied by a decrease in MT dynamics. These results suggest that the relationship between MT dynamics and contractility is an indirect one.
Danowski
B A
BA
Department of Biology, Union College, Schenectady, New York 12308, USA. danowskb@union.edu
D016475:3T3 Cells; D000818:Animals; D002455:Cell Division; D016895:Culture Media, Serum-Free; D004351:Drug Resistance; D005347:Fibroblasts / Q000187:drug effects / Q000502:physiology*; D051379:Mice; D008870:Microtubules / Q000187:drug effects / Q000502:physiology*; D015739:Nocodazole / Q000494:pharmacology; D012710:Serum Albumin, Bovine / Q000494:pharmacology
D016895:Culture Media, Serum-Free; D012710:Serum Albumin, Bovine; D015739:Nocodazole