TY - JOUR
T1 - TRPM7 triggers Ca2+ sparks and invadosome formation in neuroblastoma cells
AU - Visser, Daan
AU - Langeslag, Michiel
AU - Kedziora, Katarzyna M
AU - Klarenbeek, Jeffrey
AU - Kamermans, Alwin
AU - Horgen, F David
AU - Fleig, Andrea
AU - van Leeuwen, Frank N
AU - Jalink, Kees
N1 - Copyright © 2013 Elsevier Ltd. All rights reserved.
PY - 2013/12
Y1 - 2013/12
N2 - Cell migration depends on the dynamic formation and turnover of cell adhesions and is tightly controlled by actomyosin contractility and local Ca2+ signals. The divalent cation channel TRPM7 (Transient Receptor Potential cation channel, subfamily Melastatin, member 7) has recently received much attention as a regulator of cell adhesion, migration and (localized) Ca2+ signaling. Overexpression and knockdown of TRPM7 affects actomyosin contractility and the formation of cell adhesions such as invadosomes and focal adhesions, but the role of TRPM7-mediated Ca2+ signals herein is currently not understood. Using Total Internal Reflection Fluorescence (TIRF) Ca2+ fluorometry and a novel automated analysis routine we have addressed the role of Ca2+ in the control of invadosome dynamics in N1E-115 mouse neuroblastoma cells. We find that TRPM7 promotes the formation of highly repetitive and localized Ca2+ microdomains or "Ca2+ sparking hotspots" at the ventral plasma membrane. Ca2+ sparking appears strictly dependent on extracellular Ca2+ and is abolished by TRPM7 channel inhibitors such as waixenicin-A. TRPM7 inhibition also induces invadosome dissolution. However, invadosome formation is (functionally and spatially) dissociated from TRPM7-mediated Ca2+ sparks. Rather, our data indicate that TRPM7 affects actomyosin contractility and invadosome formation independent of Ca2+ influx.
AB - Cell migration depends on the dynamic formation and turnover of cell adhesions and is tightly controlled by actomyosin contractility and local Ca2+ signals. The divalent cation channel TRPM7 (Transient Receptor Potential cation channel, subfamily Melastatin, member 7) has recently received much attention as a regulator of cell adhesion, migration and (localized) Ca2+ signaling. Overexpression and knockdown of TRPM7 affects actomyosin contractility and the formation of cell adhesions such as invadosomes and focal adhesions, but the role of TRPM7-mediated Ca2+ signals herein is currently not understood. Using Total Internal Reflection Fluorescence (TIRF) Ca2+ fluorometry and a novel automated analysis routine we have addressed the role of Ca2+ in the control of invadosome dynamics in N1E-115 mouse neuroblastoma cells. We find that TRPM7 promotes the formation of highly repetitive and localized Ca2+ microdomains or "Ca2+ sparking hotspots" at the ventral plasma membrane. Ca2+ sparking appears strictly dependent on extracellular Ca2+ and is abolished by TRPM7 channel inhibitors such as waixenicin-A. TRPM7 inhibition also induces invadosome dissolution. However, invadosome formation is (functionally and spatially) dissociated from TRPM7-mediated Ca2+ sparks. Rather, our data indicate that TRPM7 affects actomyosin contractility and invadosome formation independent of Ca2+ influx.
KW - Acetates/pharmacology
KW - Actomyosin/metabolism
KW - Animals
KW - Calcium/metabolism
KW - Calcium Signaling
KW - Cell Adhesion/drug effects
KW - Cell Line, Tumor
KW - Cell Membrane/metabolism
KW - Cell Movement/drug effects
KW - Diterpenes/pharmacology
KW - Mice
KW - Neuroblastoma/metabolism
KW - RNA Interference
KW - RNA, Small Interfering/metabolism
KW - TRPM Cation Channels/antagonists & inhibitors
U2 - 10.1016/j.ceca.2013.09.003
DO - 10.1016/j.ceca.2013.09.003
M3 - Article
C2 - 24176224
VL - 54
SP - 404
EP - 415
JO - Cell calcium
JF - Cell calcium
SN - 0143-4160
IS - 6
ER -