ROCK activity and the Gβγ complex mediate chemotactic migration of mouse bone marrow-derived stromal cells

Files in This Item:
File Description SizeFormat 
ROCK activity and the GBY complex mediatie chemotactic migration of mouse bone marrow...pdf3.67 MBAdobe PDFDownload
Title: ROCK activity and the Gβγ complex mediate chemotactic migration of mouse bone marrow-derived stromal cells
Authors: Ryan, Caroline M.
Brown, James A. L.
Bourke, Emer
Prendergast, Áine M.
Kavanagh, Claire
Liu, Zhonglin
Owens, Peter
Shaw, Georgina
Kolch, Walter
O'Brien, Timothy
Barry, Frank P.
Permanent link: http://hdl.handle.net/10197/9159
Date: 24-Jul-2015
Abstract: Bone marrow-derived stromal cells (BMSCs), also known as mesenchymal stem cells, are the focus of intensive efforts worldwide to elucidate their function and biology. Despite the importance of BMSC migration for their potential therapeutic uses, the mechanisms and signalling governing stem cell migration are still not fully elucidated. Methods: We investigated and detailed the effects of MCP-1 activation on BMSCs by using inhibitors of G protein-coupled receptor alpha beta (GPCR αβ), ROCK (Rho-associated, coiled-coil containing protein kinase), and PI3 kinase (PI3K). The effects of MCP-1 stimulation on intracellular signalling cascades were characterised by using immunoblotting and immunofluorescence. The effectors of MCP-1-mediated migration were investigated by using migration assays (both two-dimensional and three-dimensional) in combination with inhibitors. Results: We established the kinetics of the MCP-1-activated signalling cascade and show that this cascade correlates with cell surface re-localisation of chemokine (C motif) receptor 2 (CCR2) (the MCP-1 receptor) to the cell periphery following MCP-1 stimulation. We show that MCP-1-initiated signalling is dependent on the activation of βγ subunits from the GPCR αβγ complex. In addition, we characterise a novel role for PI3Kγ signalling for the activation of both PAK and ERK following MCP-1 stimulation. We present evidence that the Gβγ complex is responsible for PI3K/Akt, PAK, and ERK signalling induced by MCP-1 in BMSCs. Importantly, we found that, in BMSCs, inhibition of ROCK significantly inhibits MCP-1-induced chemotactic migration, in contrast to previous reports in other systems.Conclusions: Our results indicate differential chemotactic signalling in mouse BMSCs, which has important implications for the translation of in vivo mouse model findings into human trials. We identified novel components and interactions activated by MCP-1-mediated signalling, which are important for stem cell migration. This work has identified additional potential therapeutic targets that could be manipulated to improve BMSC delivery and homing.
Funding Details: Science Foundation Ireland
Type of material: Journal Article
Publisher: Springer Nature
Copyright (published version): 2015 the Authors
Keywords: ROCKGβγ complexMesenchymal stem cellsBMSCsMCP-1 activationMCP-1 activationGPCR αβChemotactic migrationChemotactic signallingStem cell migration
DOI: 10.1186/s13287-015-0125-y
Language: en
Status of Item: Peer reviewed
Appears in Collections:SBI Research Collection

Show full item record

SCOPUSTM   
Citations 50

6
Last Week
0
Last month
checked on Aug 17, 2018

Google ScholarTM

Check

Altmetric


This item is available under the Attribution-NonCommercial-NoDerivs 3.0 Ireland. No item may be reproduced for commercial purposes. For other possible restrictions on use please refer to the publisher's URL where this is made available, or to notes contained in the item itself. Other terms may apply.