Activation of cytosolic phospholipase A2 downstream of the Src-phospholipase D1 (PLD1)-Protein kinase C γ (PKCγ) signaling axis is required for hypoxia-induced pathological retinal angiogenesis

Qiuhua Zhang, Dong Wang, Nikhlesh Singh, Venkatesh Kundumani-Sridharan, Laxmisilpa Gadiparthi, Ch Mohan Rao, Rao Gadiparthi

Research output: Contribution to journalArticle

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Abstract

In view of understanding the mechanisms of retinal neovascularization, we had reported previously that vascular endothelial growth factor (VEGF)-induced pathological retinal angiogenesis requires the activation of Src-PLD1-PKCγ signaling. In the present work, we have identified cytosolic phospholipase A2 (cPLA2) as an effector molecule of Src-PLD1-PKCγ signaling in the mediation of VEGF-induced pathological retinal angiogenesis based on the following observations. VEGF induced cPLA2 phosphorylation in a time-dependent manner in human retinal microvascular endothelial cells (HRMVECs). VEGF also induced arachidonic acid (AA) release in a dose-, time-, and cPLA2-dependent manner. Depletion of cPLA 2 levels inhibited VEGF-induced HRMVEC DNA synthesis, migration, and tube formation. In addition, the exogenous addition of AA rescued VEGF-induced HRMVEC DNA synthesis, migration, and tube formation from inhibition by down-regulation of cPLA2. Inhibition of Src, PLD1, or PKCγ attenuated VEGF-induced cPLA2 phosphorylation and AA release. Consistent with these findings, hypoxia induced cPLA2 phosphorylation and activity in VEGF-Src-PLD1-PKCγ-dependent manner in a mouse model of oxygen-induced retinopathy. In addition, siRNA-mediated down-regulation of cPLA2 levels in the retina abrogated hypoxia-induced retinal endothelial cell proliferation and neovascularization. These observations suggest that cPLA2-dependent AA release is required for VEGF-induced Src-PLD1-PKCγ-mediated pathological retinal angiogenesis.

Original languageEnglish (US)
Pages (from-to)22489-22498
Number of pages10
JournalJournal of Biological Chemistry
Volume286
Issue number25
DOIs
StatePublished - Jun 24 2011

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Pathologic Neovascularization
Cytosolic Phospholipases A2
Protein Kinase C
Vascular Endothelial Growth Factor A
Chemical activation
Endothelial cells
Arachidonic Acid
Phosphorylation
Endothelial Cells
Down-Regulation
Retinal Neovascularization
Hypoxia
phospholipase D1
DNA
Cell proliferation
Small Interfering RNA
Retina
Cell Proliferation
Oxygen

All Science Journal Classification (ASJC) codes

  • Biochemistry
  • Molecular Biology
  • Cell Biology

Cite this

Activation of cytosolic phospholipase A2 downstream of the Src-phospholipase D1 (PLD1)-Protein kinase C γ (PKCγ) signaling axis is required for hypoxia-induced pathological retinal angiogenesis. / Zhang, Qiuhua; Wang, Dong; Singh, Nikhlesh; Kundumani-Sridharan, Venkatesh; Gadiparthi, Laxmisilpa; Rao, Ch Mohan; Gadiparthi, Rao.

In: Journal of Biological Chemistry, Vol. 286, No. 25, 24.06.2011, p. 22489-22498.

Research output: Contribution to journalArticle

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abstract = "In view of understanding the mechanisms of retinal neovascularization, we had reported previously that vascular endothelial growth factor (VEGF)-induced pathological retinal angiogenesis requires the activation of Src-PLD1-PKCγ signaling. In the present work, we have identified cytosolic phospholipase A2 (cPLA2) as an effector molecule of Src-PLD1-PKCγ signaling in the mediation of VEGF-induced pathological retinal angiogenesis based on the following observations. VEGF induced cPLA2 phosphorylation in a time-dependent manner in human retinal microvascular endothelial cells (HRMVECs). VEGF also induced arachidonic acid (AA) release in a dose-, time-, and cPLA2-dependent manner. Depletion of cPLA 2 levels inhibited VEGF-induced HRMVEC DNA synthesis, migration, and tube formation. In addition, the exogenous addition of AA rescued VEGF-induced HRMVEC DNA synthesis, migration, and tube formation from inhibition by down-regulation of cPLA2. Inhibition of Src, PLD1, or PKCγ attenuated VEGF-induced cPLA2 phosphorylation and AA release. Consistent with these findings, hypoxia induced cPLA2 phosphorylation and activity in VEGF-Src-PLD1-PKCγ-dependent manner in a mouse model of oxygen-induced retinopathy. In addition, siRNA-mediated down-regulation of cPLA2 levels in the retina abrogated hypoxia-induced retinal endothelial cell proliferation and neovascularization. These observations suggest that cPLA2-dependent AA release is required for VEGF-induced Src-PLD1-PKCγ-mediated pathological retinal angiogenesis.",
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