Capillary pericytes express α-smooth muscle actin, which requires prevention of filamentous-actin depolymerization for detection

Elife. 2018 Mar 21:7:e34861. doi: 10.7554/eLife.34861.

Abstract

Recent evidence suggests that capillary pericytes are contractile and play a crucial role in the regulation of microcirculation. However, failure to detect components of the contractile apparatus in capillary pericytes, most notably α-smooth muscle actin (α-SMA), has questioned these findings. Using strategies that allow rapid filamentous-actin (F-actin) fixation (i.e. snap freeze fixation with methanol at -20°C) or prevent F-actin depolymerization (i.e. with F-actin stabilizing agents), we demonstrate that pericytes on mouse retinal capillaries, including those in intermediate and deeper plexus, express α-SMA. Junctional pericytes were more frequently α-SMA-positive relative to pericytes on linear capillary segments. Intravitreal administration of short interfering RNA (α-SMA-siRNA) suppressed α-SMA expression preferentially in high order branch capillary pericytes, confirming the existence of a smaller pool of α-SMA in distal capillary pericytes that is quickly lost by depolymerization. We conclude that capillary pericytes do express α-SMA, which rapidly depolymerizes during tissue fixation thus evading detection by immunolabeling.

Keywords: F-actin; alpha-smooth muscle actin; capillary; mouse; neuroscience; retinal pericytes.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Actins / genetics
  • Actins / metabolism*
  • Animals
  • Capillaries / cytology
  • Capillaries / metabolism*
  • Immunohistochemistry
  • Mice, Transgenic
  • Muscle, Smooth / metabolism
  • Pericytes / metabolism*
  • Polymerization
  • RNA Interference
  • Retinal Vessels / metabolism*

Substances

  • Acta2 protein, mouse
  • Actins