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Polarized migration of lymphatic endothelial cells is critically dependent on podoplanin regulation of Cdc42
- Source :
- American journal of physiology. Lung cellular and molecular physiology. 300(1)
- Publication Year :
- 2010
-
Abstract
- We have shown previously that T1α/podoplanin is required for capillary tube formation by human lung microvascular lymphatic endothelial cells (HMVEC-LLy) and that cells with decreased podoplanin expression fail to properly activate the small GTPase RhoA shortly after the beginning of the lymphangiogenic process. The objective of this study was to determine whether podoplanin regulates HMVEC-LLy migration and whether this regulation is via modulation of small GTPase activation. In analysis of scratch wound assays, we found that small interfering RNA (siRNA) depletion of podoplanin expression in HMVEC-LLy inhibits VEGF-induced microtubule-organizing center (MTOC) and Golgi polarization and causes a dramatic reduction in directional migration compared with control siRNA-transfected cells. In addition, a striking redistribution of cortical actin to fiber networks across the cell body is observed in these cells, and, remarkably, it returns to control levels if the cells are cotransfected with a dominant-negative mutant of Cdc42. Moreover, cotransfection of a dominant-negative construct of Cdc42 into podoplanin knockdown HMVEC-LLy completely abrogated the effect of podoplanin deficiency, rescuing MTOC and Golgi polarization and cell migration to control level. Importantly, expression of constitutively active Cdc42 construct, like podoplanin knockdown, decreased RhoA-GTP level in HMVEC-LLy, demonstrating cross talk between both GTPases. Taken together, the results indicate that polarized migration of lymphatic endothelial cells in response to VEGF is mediated via a pathway of podoplanin regulation of small GTPase activities, in particular Cdc42.
- Subjects :
- Pulmonary and Respiratory Medicine
Small interfering RNA
RHOA
Physiology
Golgi Apparatus
CDC42
GTP Phosphohydrolases
Cell Movement
Physiology (medical)
Cell polarity
Humans
Small GTPase
cdc42 GTP-Binding Protein
Lung
Monomeric GTP-Binding Proteins
Membrane Glycoproteins
biology
Microcirculation
Endothelial Cells
Cell migration
Cell Biology
Cell biology
Endothelial stem cell
Podoplanin
biology.protein
GTP Phosphohydrolase Activators
rhoA GTP-Binding Protein
Subjects
Details
- ISSN :
- 15221504
- Volume :
- 300
- Issue :
- 1
- Database :
- OpenAIRE
- Journal :
- American journal of physiology. Lung cellular and molecular physiology
- Accession number :
- edsair.doi.dedup.....59100f8f895a5def6c431309b9c4b506