Deficiencies of the Lipid-Signaling Enzymes Phospholipase D1 and D2 Alter Cytoskeletal Organization, Macrophage Phagocytosis, and Cytokine-Stimulated Neutrophil Recruitment

Wahida H. Ali, Qin Chen, Kathleen E. Delgiorno, Wenjuan Su, Jason C. Hall, Tsunaki Hongu, Huasong Tian, Yasunori Kanaho, Gilbert Di Paolo, Howard C. Crawford, Michael A. Frohman

Research output: Contribution to journalArticle

38 Citations (Scopus)

Abstract

Cell migration and phagocytosis ensue from extracellular-initiated signaling cascades that orchestrate dynamic reorganization of the actin cytoskeleton. The reorganization is mediated by effector proteins recruited to the site of activity by locally-generated lipid second messengers. Phosphatidic acid (PA), a membrane phospholipid generated by multiple enzyme families including Phospholipase D (PLD), has been proposed to function in this role. Here, we show that macrophages prepared from mice lacking either of the classical PLD isoforms PLD1 or PLD2, or wild-type macrophages whose PLD activity has been pharmacologically inhibited, display isoform-specific actin cytoskeleton abnormalities that likely underlie decreases observed in phagocytic capacity. Unexpectedly, PA continued to be detected on the phagosome in the absence of either isoform and even when all PLD activity was eliminated. However, a disorganized phagocytic cup was observed as visualized by imaging PA, F-actin, Rac1, an organizer of the F-actin network, and DOCK2, a Rac1 activator, suggesting that PLD-mediated PA production during phagocytosis is specifically critical for the integrity of the process. The abnormal F-actin reorganization additionally impacted neutrophil migration and extravasation from the vasculature into interstitial tissues. Although both PLD1 and PLD2 were important in these processes, we also observed isoform-specific functions. PLD1-driven processes in particular were observed to be critical in transmigration of macrophages exiting the vasculature during immune responses such as those seen in acute pancreatitis or irritant-induced skin vascularization.

Original languageEnglish (US)
Article numbere55325
JournalPLoS One
Volume8
Issue number1
DOIs
StatePublished - Jan 28 2013

Fingerprint

Phospholipase D
phospholipase D
Neutrophil Infiltration
Macrophages
phagocytosis
Phosphatidic Acids
Phagocytosis
neutrophils
Actins
macrophages
cytokines
Cytokines
Lipids
Protein Isoforms
actin
Enzymes
lipids
enzymes
acids
microfilaments

ASJC Scopus subject areas

  • Agricultural and Biological Sciences(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Medicine(all)

Cite this

Deficiencies of the Lipid-Signaling Enzymes Phospholipase D1 and D2 Alter Cytoskeletal Organization, Macrophage Phagocytosis, and Cytokine-Stimulated Neutrophil Recruitment. / Ali, Wahida H.; Chen, Qin; Delgiorno, Kathleen E.; Su, Wenjuan; Hall, Jason C.; Hongu, Tsunaki; Tian, Huasong; Kanaho, Yasunori; Di Paolo, Gilbert; Crawford, Howard C.; Frohman, Michael A.

In: PLoS One, Vol. 8, No. 1, e55325, 28.01.2013.

Research output: Contribution to journalArticle

Ali, WH, Chen, Q, Delgiorno, KE, Su, W, Hall, JC, Hongu, T, Tian, H, Kanaho, Y, Di Paolo, G, Crawford, HC & Frohman, MA 2013, 'Deficiencies of the Lipid-Signaling Enzymes Phospholipase D1 and D2 Alter Cytoskeletal Organization, Macrophage Phagocytosis, and Cytokine-Stimulated Neutrophil Recruitment', PLoS One, vol. 8, no. 1, e55325. https://doi.org/10.1371/journal.pone.0055325
Ali, Wahida H. ; Chen, Qin ; Delgiorno, Kathleen E. ; Su, Wenjuan ; Hall, Jason C. ; Hongu, Tsunaki ; Tian, Huasong ; Kanaho, Yasunori ; Di Paolo, Gilbert ; Crawford, Howard C. ; Frohman, Michael A. / Deficiencies of the Lipid-Signaling Enzymes Phospholipase D1 and D2 Alter Cytoskeletal Organization, Macrophage Phagocytosis, and Cytokine-Stimulated Neutrophil Recruitment. In: PLoS One. 2013 ; Vol. 8, No. 1.
@article{022bb3f05e614baabc46d2ce5e36ed55,
title = "Deficiencies of the Lipid-Signaling Enzymes Phospholipase D1 and D2 Alter Cytoskeletal Organization, Macrophage Phagocytosis, and Cytokine-Stimulated Neutrophil Recruitment",
abstract = "Cell migration and phagocytosis ensue from extracellular-initiated signaling cascades that orchestrate dynamic reorganization of the actin cytoskeleton. The reorganization is mediated by effector proteins recruited to the site of activity by locally-generated lipid second messengers. Phosphatidic acid (PA), a membrane phospholipid generated by multiple enzyme families including Phospholipase D (PLD), has been proposed to function in this role. Here, we show that macrophages prepared from mice lacking either of the classical PLD isoforms PLD1 or PLD2, or wild-type macrophages whose PLD activity has been pharmacologically inhibited, display isoform-specific actin cytoskeleton abnormalities that likely underlie decreases observed in phagocytic capacity. Unexpectedly, PA continued to be detected on the phagosome in the absence of either isoform and even when all PLD activity was eliminated. However, a disorganized phagocytic cup was observed as visualized by imaging PA, F-actin, Rac1, an organizer of the F-actin network, and DOCK2, a Rac1 activator, suggesting that PLD-mediated PA production during phagocytosis is specifically critical for the integrity of the process. The abnormal F-actin reorganization additionally impacted neutrophil migration and extravasation from the vasculature into interstitial tissues. Although both PLD1 and PLD2 were important in these processes, we also observed isoform-specific functions. PLD1-driven processes in particular were observed to be critical in transmigration of macrophages exiting the vasculature during immune responses such as those seen in acute pancreatitis or irritant-induced skin vascularization.",
author = "Ali, {Wahida H.} and Qin Chen and Delgiorno, {Kathleen E.} and Wenjuan Su and Hall, {Jason C.} and Tsunaki Hongu and Huasong Tian and Yasunori Kanaho and {Di Paolo}, Gilbert and Crawford, {Howard C.} and Frohman, {Michael A.}",
year = "2013",
month = "1",
day = "28",
doi = "10.1371/journal.pone.0055325",
language = "English (US)",
volume = "8",
journal = "PLoS One",
issn = "1932-6203",
publisher = "Public Library of Science",
number = "1",

}

TY - JOUR

T1 - Deficiencies of the Lipid-Signaling Enzymes Phospholipase D1 and D2 Alter Cytoskeletal Organization, Macrophage Phagocytosis, and Cytokine-Stimulated Neutrophil Recruitment

AU - Ali, Wahida H.

AU - Chen, Qin

AU - Delgiorno, Kathleen E.

AU - Su, Wenjuan

AU - Hall, Jason C.

AU - Hongu, Tsunaki

AU - Tian, Huasong

AU - Kanaho, Yasunori

AU - Di Paolo, Gilbert

AU - Crawford, Howard C.

AU - Frohman, Michael A.

PY - 2013/1/28

Y1 - 2013/1/28

N2 - Cell migration and phagocytosis ensue from extracellular-initiated signaling cascades that orchestrate dynamic reorganization of the actin cytoskeleton. The reorganization is mediated by effector proteins recruited to the site of activity by locally-generated lipid second messengers. Phosphatidic acid (PA), a membrane phospholipid generated by multiple enzyme families including Phospholipase D (PLD), has been proposed to function in this role. Here, we show that macrophages prepared from mice lacking either of the classical PLD isoforms PLD1 or PLD2, or wild-type macrophages whose PLD activity has been pharmacologically inhibited, display isoform-specific actin cytoskeleton abnormalities that likely underlie decreases observed in phagocytic capacity. Unexpectedly, PA continued to be detected on the phagosome in the absence of either isoform and even when all PLD activity was eliminated. However, a disorganized phagocytic cup was observed as visualized by imaging PA, F-actin, Rac1, an organizer of the F-actin network, and DOCK2, a Rac1 activator, suggesting that PLD-mediated PA production during phagocytosis is specifically critical for the integrity of the process. The abnormal F-actin reorganization additionally impacted neutrophil migration and extravasation from the vasculature into interstitial tissues. Although both PLD1 and PLD2 were important in these processes, we also observed isoform-specific functions. PLD1-driven processes in particular were observed to be critical in transmigration of macrophages exiting the vasculature during immune responses such as those seen in acute pancreatitis or irritant-induced skin vascularization.

AB - Cell migration and phagocytosis ensue from extracellular-initiated signaling cascades that orchestrate dynamic reorganization of the actin cytoskeleton. The reorganization is mediated by effector proteins recruited to the site of activity by locally-generated lipid second messengers. Phosphatidic acid (PA), a membrane phospholipid generated by multiple enzyme families including Phospholipase D (PLD), has been proposed to function in this role. Here, we show that macrophages prepared from mice lacking either of the classical PLD isoforms PLD1 or PLD2, or wild-type macrophages whose PLD activity has been pharmacologically inhibited, display isoform-specific actin cytoskeleton abnormalities that likely underlie decreases observed in phagocytic capacity. Unexpectedly, PA continued to be detected on the phagosome in the absence of either isoform and even when all PLD activity was eliminated. However, a disorganized phagocytic cup was observed as visualized by imaging PA, F-actin, Rac1, an organizer of the F-actin network, and DOCK2, a Rac1 activator, suggesting that PLD-mediated PA production during phagocytosis is specifically critical for the integrity of the process. The abnormal F-actin reorganization additionally impacted neutrophil migration and extravasation from the vasculature into interstitial tissues. Although both PLD1 and PLD2 were important in these processes, we also observed isoform-specific functions. PLD1-driven processes in particular were observed to be critical in transmigration of macrophages exiting the vasculature during immune responses such as those seen in acute pancreatitis or irritant-induced skin vascularization.

UR - http://www.scopus.com/inward/record.url?scp=84873801908&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84873801908&partnerID=8YFLogxK

U2 - 10.1371/journal.pone.0055325

DO - 10.1371/journal.pone.0055325

M3 - Article

C2 - 23383154

AN - SCOPUS:84873801908

VL - 8

JO - PLoS One

JF - PLoS One

SN - 1932-6203

IS - 1

M1 - e55325

ER -