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


Alarcon-Martinez L., Yilmaz-Ozcan S., Yemisci M., Schallek J., Kilic K., Can A., ...More

ELIFE, vol.7, 2018 (SCI-Expanded) identifier identifier identifier

  • Publication Type: Article / Article
  • Volume: 7
  • Publication Date: 2018
  • Doi Number: 10.7554/elife.34861
  • Journal Name: ELIFE
  • Journal Indexes: Science Citation Index Expanded (SCI-EXPANDED), Scopus
  • Hacettepe University Affiliated: Yes

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 alpha-smooth muscle actin (alpha-SMA), has questioned these findings. Using strategies that allow rapid filamentous-actin (F-actin) fixation (i.e. snap freeze fixation with methanol at -20 degrees 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 alpha-SMA. Junctional pericytes were more frequently alpha-SMA-positive relative to pericytes on linear capillary segments. Intravitreal administration of short interfering RNA (alpha-SMA-siRNA) suppressed alpha-SMA expression preferentially in high order branch capillary pericytes, confirming the existence of a smaller pool of alpha-SMA in distal capillary pericytes that is quickly lost by depolymerization. We conclude that capillary pericytes do express alpha-SMA, which rapidly depolymerizes during tissue fixation thus evading detection by immunolabeling.