Ca2+-dependent myosin II activation is required for uropod retraction during neutrophil migration

Robert J. Eddy, Lynda M. Pierini, Fumio Matsumura, Frederick R. Maxfield

Research output: Contribution to journalArticle

164 Citations (Scopus)

Abstract

Buffering of intracellular Ca2+ transients in human neutrophils leads to reduced motility due to defective uropod detachment on fibronectin and vitronectin-coated surfaces. Since one potential target of a rise in [Ca2+](i) is the activation of myosin II, we characterized the role of myosin II during motility. Treatment of neutrophils with a myosin inhibitor (2,3-butanedione monoxime), or myosin light chain kinase inhibitors (ML-7, ML-9, or KT5926) resulted in impaired uropod retraction and a dose-dependent decrease in chemokinesis following stimulation with N-formyl-Met-Leu-Phe (fMLP). Treatment with ML-9 resulted in a redistribution of F-actin and talin to the non-retracted uropods, mimicking the redistribution observed during [Ca2+](i) buffering. Impairment of uropod retraction and redistribution of F-actin and talin by myosin II inhibition was only observed on adhesive substrates such as fibronectin and not on poorly adhesive substrates such as human serum-coated glass. At higher concentrations of ML-9, cell polarization was inhibited and pseudopod extension occurred radially. Using an antibody specific for serine 19-phosphorylated regulatory light chain of myosin II, regions of activated myosin II were found at the leading edge as well as the uropod in motile fMLP-stimulated cells. [Ca2+](i) depletion caused a 50% decrease in the level of serine 19-phosphorylated myosin II suggesting that activation of myosin II by intracellular Ca2+ transients may be an essential step in establishing a polarized pseudopod and providing the force required for uropod retraction during PMN motility on adhesive surfaces.

Original languageEnglish (US)
Pages (from-to)1287-1298
Number of pages12
JournalJournal of Cell Science
Volume113
Issue number7
StatePublished - 2000
Externally publishedYes

Fingerprint

Myosin Type II
Neutrophils
Talin
Adhesives
Pseudopodia
Fibronectins
Serine
Actins
Myosin-Light-Chain Kinase
Vitronectin
Myosins
Glass
Light
Antibodies
Serum

Keywords

  • Motility
  • Myosin II
  • Neutrophil

ASJC Scopus subject areas

  • Cell Biology

Cite this

Ca2+-dependent myosin II activation is required for uropod retraction during neutrophil migration. / Eddy, Robert J.; Pierini, Lynda M.; Matsumura, Fumio; Maxfield, Frederick R.

In: Journal of Cell Science, Vol. 113, No. 7, 2000, p. 1287-1298.

Research output: Contribution to journalArticle

Eddy, RJ, Pierini, LM, Matsumura, F & Maxfield, FR 2000, 'Ca2+-dependent myosin II activation is required for uropod retraction during neutrophil migration', Journal of Cell Science, vol. 113, no. 7, pp. 1287-1298.
Eddy, Robert J. ; Pierini, Lynda M. ; Matsumura, Fumio ; Maxfield, Frederick R. / Ca2+-dependent myosin II activation is required for uropod retraction during neutrophil migration. In: Journal of Cell Science. 2000 ; Vol. 113, No. 7. pp. 1287-1298.
@article{a288a47d702d4139b43afe4d128a0ff8,
title = "Ca2+-dependent myosin II activation is required for uropod retraction during neutrophil migration",
abstract = "Buffering of intracellular Ca2+ transients in human neutrophils leads to reduced motility due to defective uropod detachment on fibronectin and vitronectin-coated surfaces. Since one potential target of a rise in [Ca2+](i) is the activation of myosin II, we characterized the role of myosin II during motility. Treatment of neutrophils with a myosin inhibitor (2,3-butanedione monoxime), or myosin light chain kinase inhibitors (ML-7, ML-9, or KT5926) resulted in impaired uropod retraction and a dose-dependent decrease in chemokinesis following stimulation with N-formyl-Met-Leu-Phe (fMLP). Treatment with ML-9 resulted in a redistribution of F-actin and talin to the non-retracted uropods, mimicking the redistribution observed during [Ca2+](i) buffering. Impairment of uropod retraction and redistribution of F-actin and talin by myosin II inhibition was only observed on adhesive substrates such as fibronectin and not on poorly adhesive substrates such as human serum-coated glass. At higher concentrations of ML-9, cell polarization was inhibited and pseudopod extension occurred radially. Using an antibody specific for serine 19-phosphorylated regulatory light chain of myosin II, regions of activated myosin II were found at the leading edge as well as the uropod in motile fMLP-stimulated cells. [Ca2+](i) depletion caused a 50{\%} decrease in the level of serine 19-phosphorylated myosin II suggesting that activation of myosin II by intracellular Ca2+ transients may be an essential step in establishing a polarized pseudopod and providing the force required for uropod retraction during PMN motility on adhesive surfaces.",
keywords = "Motility, Myosin II, Neutrophil",
author = "Eddy, {Robert J.} and Pierini, {Lynda M.} and Fumio Matsumura and Maxfield, {Frederick R.}",
year = "2000",
language = "English (US)",
volume = "113",
pages = "1287--1298",
journal = "Journal of Cell Science",
issn = "0021-9533",
publisher = "Company of Biologists Ltd",
number = "7",

}

TY - JOUR

T1 - Ca2+-dependent myosin II activation is required for uropod retraction during neutrophil migration

AU - Eddy, Robert J.

AU - Pierini, Lynda M.

AU - Matsumura, Fumio

AU - Maxfield, Frederick R.

PY - 2000

Y1 - 2000

N2 - Buffering of intracellular Ca2+ transients in human neutrophils leads to reduced motility due to defective uropod detachment on fibronectin and vitronectin-coated surfaces. Since one potential target of a rise in [Ca2+](i) is the activation of myosin II, we characterized the role of myosin II during motility. Treatment of neutrophils with a myosin inhibitor (2,3-butanedione monoxime), or myosin light chain kinase inhibitors (ML-7, ML-9, or KT5926) resulted in impaired uropod retraction and a dose-dependent decrease in chemokinesis following stimulation with N-formyl-Met-Leu-Phe (fMLP). Treatment with ML-9 resulted in a redistribution of F-actin and talin to the non-retracted uropods, mimicking the redistribution observed during [Ca2+](i) buffering. Impairment of uropod retraction and redistribution of F-actin and talin by myosin II inhibition was only observed on adhesive substrates such as fibronectin and not on poorly adhesive substrates such as human serum-coated glass. At higher concentrations of ML-9, cell polarization was inhibited and pseudopod extension occurred radially. Using an antibody specific for serine 19-phosphorylated regulatory light chain of myosin II, regions of activated myosin II were found at the leading edge as well as the uropod in motile fMLP-stimulated cells. [Ca2+](i) depletion caused a 50% decrease in the level of serine 19-phosphorylated myosin II suggesting that activation of myosin II by intracellular Ca2+ transients may be an essential step in establishing a polarized pseudopod and providing the force required for uropod retraction during PMN motility on adhesive surfaces.

AB - Buffering of intracellular Ca2+ transients in human neutrophils leads to reduced motility due to defective uropod detachment on fibronectin and vitronectin-coated surfaces. Since one potential target of a rise in [Ca2+](i) is the activation of myosin II, we characterized the role of myosin II during motility. Treatment of neutrophils with a myosin inhibitor (2,3-butanedione monoxime), or myosin light chain kinase inhibitors (ML-7, ML-9, or KT5926) resulted in impaired uropod retraction and a dose-dependent decrease in chemokinesis following stimulation with N-formyl-Met-Leu-Phe (fMLP). Treatment with ML-9 resulted in a redistribution of F-actin and talin to the non-retracted uropods, mimicking the redistribution observed during [Ca2+](i) buffering. Impairment of uropod retraction and redistribution of F-actin and talin by myosin II inhibition was only observed on adhesive substrates such as fibronectin and not on poorly adhesive substrates such as human serum-coated glass. At higher concentrations of ML-9, cell polarization was inhibited and pseudopod extension occurred radially. Using an antibody specific for serine 19-phosphorylated regulatory light chain of myosin II, regions of activated myosin II were found at the leading edge as well as the uropod in motile fMLP-stimulated cells. [Ca2+](i) depletion caused a 50% decrease in the level of serine 19-phosphorylated myosin II suggesting that activation of myosin II by intracellular Ca2+ transients may be an essential step in establishing a polarized pseudopod and providing the force required for uropod retraction during PMN motility on adhesive surfaces.

KW - Motility

KW - Myosin II

KW - Neutrophil

UR - http://www.scopus.com/inward/record.url?scp=0034107669&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0034107669&partnerID=8YFLogxK

M3 - Article

C2 - 10704379

AN - SCOPUS:0034107669

VL - 113

SP - 1287

EP - 1298

JO - Journal of Cell Science

JF - Journal of Cell Science

SN - 0021-9533

IS - 7

ER -