Cytoskeleton dynamics in drug-treated platelets

Anal Bioanal Chem. 2015 Apr;407(10):2803-9. doi: 10.1007/s00216-015-8523-7. Epub 2015 Feb 21.

Abstract

Platelet activation is a key process in blood clot formation. During activation, platelets go through both chemical and physical changes, including secretion of chemical messengers and cellular shape change. Platelet shape change is mediated by the two major cytoskeletal elements in platelets, the actin matrix and microtubule ring. Most studies to date have evaluated these structures qualitatively, whereas this paper aims to provide a quantitative method of examining changes in these structures by fluorescently labeling the element of interest and performing single cell image analysis. The method described herein tracks the diameter of the microtubule ring and the circumference of the actin matrix as they change over time. Platelets were incubated with a series of drugs that interact with tubulin or actin, and the platelets were observed for variation in shape change dynamics throughout the activation process. Differences in shape change mechanics due to drug incubation were observable in each case.

MeSH terms

  • Actins / metabolism
  • Actins / ultrastructure
  • Animals
  • Blood Platelets / cytology*
  • Blood Platelets / drug effects*
  • Bridged Bicyclo Compounds, Heterocyclic / pharmacology
  • Cytochalasin D / pharmacology
  • Cytoskeleton / drug effects*
  • Dimethyl Sulfoxide / pharmacology
  • Fluorescent Antibody Technique / methods
  • Image Processing, Computer-Assisted
  • Microtubules / metabolism
  • Microtubules / ultrastructure
  • Paclitaxel / pharmacology
  • Rabbits
  • Thiazolidines / pharmacology
  • Vincristine / pharmacology

Substances

  • Actins
  • Bridged Bicyclo Compounds, Heterocyclic
  • Thiazolidines
  • Cytochalasin D
  • Vincristine
  • Paclitaxel
  • latrunculin A
  • Dimethyl Sulfoxide