Unloaded shortening velocity in single permeabilized vascular smooth muscle cells is independent of microtubule status.
Microtubules may influence smooth muscle contraction either via involvement in signal transduction processes or by serving as an internal load that opposes contraction. To test the latter hypothesis, microtubule distribution and the unloaded shortening velocity were investigated in freshly isolated single vascular smooth muscle cells (VSMCs) treated with microtubule modulating drugs. Immunocytochemical studies showed that microtubules run mainly longitudinally in relaxed VSMCs. They are oriented more obliquely, almost transversely to the long axis of the cells after contraction, suggesting that microtubules are compressed during shortening, and thus might impart an internal passive load. Quantitative immunocytochemical analysis revealed that, relative to the control group, colchicine (15 microM) decreased the microtubule density by 40% while taxol (10 microM) increased the microtubule density by 46%. However, alteration of microtubule polymerization status by these microtubule-modulating drugs did not have a significant effect on unloaded shortening velocity in alpha-toxin permeabilized VSMCs under maximal activating conditions or submaximal activating conditions (about 36% of maximal velocity). These results suggest that microtubules do not present an appreciable internal load to dampen single VSMCs shortening in the present experimental system, and that their influence on smooth muscle contraction is primarily via signal transduction mechanisms.
Zhang
Dahua
D
Department of Anatomy and Cell Biology, Indiana University School of Medicine, Indianapolis, IN 46202, USA.
Sherwood
Jennifer
J
Li
Liang
L
Swartz
Darl R
DR
D000199:Actins / Q000378:metabolism; D000818:Animals; D002118:Calcium / Q000494:pharmacology; D002448:Cell Adhesion / Q000502:physiology; D002463:Cell Membrane Permeability; D048430:Cell Shape / Q000187:drug effects / Q000502:physiology; D003078:Colchicine / Q000494:pharmacology; D007150:Immunohistochemistry; D007700:Kinetics; D008856:Microscopy, Fluorescence; D008870:Microtubules / Q000187:drug effects / Q000378:metabolism / Q000502:physiology*; D009131:Muscle, Smooth, Vascular / Q000166:cytology / Q000187:drug effects / Q000378:metabolism; D032389:Myocytes, Smooth Muscle / Q000187:drug effects / Q000378:metabolism / Q000502:physiology*; D015739:Nocodazole / Q000494:pharmacology; D017239:Paclitaxel / Q000494:pharmacology; D011817:Rabbits
D000199:Actins; D017239:Paclitaxel; D015739:Nocodazole; D003078:Colchicine; D002118:Calcium