Global and local regulation of clathrin-coated pit dynamics detected on patterned substrates

Biophys J. 2009 Aug 19;97(4):1038-47. doi: 10.1016/j.bpj.2009.06.003.

Abstract

Live-cell imaging of individual clathrin-coated pit (CCP) dynamics has revealed a broad variation in their internalization kinetics, but the functional significance and mechanistic underpinnings of this heterogeneity remain unknown. One contributing factor may be the spatial variations in the underlying actin cortex. To test this, we cultured cells on fibronectin (Fn) micropatterned substrates to vary the cortical actin mechanics in a defined manner. Under these conditions, stress fibers became organized to bridge adhesive islands, creating spatial heterogeneity in the cortical actin architecture. CCP lifetimes within the Fn-coated islands were selectively prolonged. This differential effect was not due to adherence to Fn-coated surfaces, and was not observed in cells grown on patterned surfaces that did not induce organized stress fiber assembly. Pharmacological agents that lower cortical tension selectively lowered CCP lifetimes within Fn islands, thus abolishing the spatial heterogeneity in the CCP dynamics. Although we cannot rule out the possibility that other factors might locally affect CCP dynamics at Fn islands, our data suggest that localized modulation in cortical tension may spatially regulate clathrin-mediated endocytosis.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Animals
  • Cell Adhesion / drug effects
  • Cell Adhesion / physiology
  • Cell Line
  • Clathrin / chemistry*
  • Clathrin / pharmacology*
  • Coated Materials, Biocompatible / chemistry*
  • Coated Materials, Biocompatible / pharmacology*
  • Epithelial Cells / drug effects
  • Epithelial Cells / physiology*
  • Haplorhini
  • Kidney / drug effects
  • Kidney / physiology*
  • Surface Properties

Substances

  • Clathrin
  • Coated Materials, Biocompatible