Cofilin mediates ATP depletion-induced endothelial cell actin alterations

Maria V. Suurna, Sharon L. Ashworth, Melanie Hosford, Ruben M. Sandoval, Sarah E. Wean, Bijal M. Shah, James R. Bamburg, Bruce Molitoris

Research output: Contribution to journalArticle

24 Citations (Scopus)

Abstract

Ischemia and sepsis lead to endothelial cell damage, resulting in compromised microvascular flow in many organs. Much remains to be determined regarding the intracellular structural events that lead to endothelial cell dysfunction. To investigate potential actin cytoskeletal-related mechanisms, ATP depletion was induced in mouse pancreatic microvascular endothelial cells (MS1). Fluorescent imaging and biochemical studies demonstrated a rapid and progressive increase in F-actin along with a decrease in G-actin at 60 min. Confocal microscopic analysis showed ATP depletion resulted in destruction of actin stress fibers and accumulation of F-actin aggregates. We hypothesized these actin alterations were secondary to dephosphorylation/activation of actin-depolymerizing factor (ADF)/cofilin proteins. Cofilin, the predominant isoform expressed in MS1 cells, was rapidly dephosphorylated/activated during ATP depletion. To directly investigate the role of cofilin activation on the actin cytoskeleton during ischemia, MS1 cells were infected with adenoviruses containing the cDNAs for wild-type Xenopus laevis ADF/cofilin green fluorescent protein [XAC(wt)-GFP], GFP, and the constitutively active and inactive isoforms XAC(S3A)-GFP and XAC(S3E)-GFP. The rate and extent of cortical actin destruction and actin aggregate formation were increased in ATP-depleted XAC(wt)-GFP- and XAC(S3A)-GFP-expressing cells, whereas increased actin stress fibers were observed in XAC(S3E)-GFP-expressing cells. To investigate the upstream signaling pathway of ADF/cofilin, LIM kinase 1-GFP (LIMK1-GFP) was expressed in MS1 cells. Cells expressing LIMK1-GFP protein had higher levels of phosphorylated ADF/cofilin, increased stress fibers, and delayed F-actin cytoskeleton destruction during ATP depletion. These results strongly support the importance of cofilin regulation in ischemia-induced endothelial cell actin cytoskeleton alterations leading to cell damage and microvascular dysfunction.

Original languageEnglish
JournalAmerican Journal of Physiology - Renal Physiology
Volume290
Issue number6
DOIs
StatePublished - Jun 2006

Fingerprint

Actin Depolymerizing Factors
Actins
Endothelial Cells
Adenosine Triphosphate
Destrin
Stress Fibers
Lim Kinases
Actin Cytoskeleton
Ischemia
Protein Isoforms
Xenopus laevis
Green Fluorescent Proteins
Adenoviridae
Sepsis
Complementary DNA

Keywords

  • Ischemia
  • Microvascular dysfunction
  • Sepsis

ASJC Scopus subject areas

  • Physiology

Cite this

Cofilin mediates ATP depletion-induced endothelial cell actin alterations. / Suurna, Maria V.; Ashworth, Sharon L.; Hosford, Melanie; Sandoval, Ruben M.; Wean, Sarah E.; Shah, Bijal M.; Bamburg, James R.; Molitoris, Bruce.

In: American Journal of Physiology - Renal Physiology, Vol. 290, No. 6, 06.2006.

Research output: Contribution to journalArticle

Suurna, Maria V. ; Ashworth, Sharon L. ; Hosford, Melanie ; Sandoval, Ruben M. ; Wean, Sarah E. ; Shah, Bijal M. ; Bamburg, James R. ; Molitoris, Bruce. / Cofilin mediates ATP depletion-induced endothelial cell actin alterations. In: American Journal of Physiology - Renal Physiology. 2006 ; Vol. 290, No. 6.
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AU - Shah, Bijal M.

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