The calcium-conducting ion channel transient receptor   potential canonical 6 is involved in macrophage   inflammatory protein-2-induced migration of mouse   neutrophils.
OBJECTIVE
The role of the calcium-conducting ion channel transient receptor potential canonical 6 (TRPC6) in macrophage inflammatory protein-2 (MIP-2) induced migration of mouse neutrophils was investigated.


METHODS
Neutrophil granulocytes isolated from murine bone marrow of wild-type (TRPC6+/+) and TRPC6 knockout (TRPC6)/)) mice were tested for the presence of TRPC6 channel expression using quantitative real-time polymerase chain reactions and immunocytochemistry. The effect of different stimuli (e.g. MIP-2, 1-oleoyl-2-acetyl-sn-glycerol, formyl-methionyl-leucyl-phenylalanin) on migration of isolated neutrophils was tested by two-dimensional (2D) migration assays, phalloidin staining and intracellular calcium imaging.


RESULTS
We found that neutrophil granulocytes express TRPC6 channels. MIP-2 induced fast cell migration of isolated neutrophils in a 2D celltracking system. Strikingly, MIP-2 was less potent in neutrophils derived from TRPC6)/) mice. These cells showed less phalloidin-coupled fluorescence and the pattern of cytosolic calcium transients was altered.


CONCLUSIONS
We describe in this paper for the first time a role for transient receptor potential (TRP) channels in migration of native lymphocytes as a new paradigm for the universal functional role of TRPs. Our data give strong evidence that TRPC6 operates downstream to CXC-type Gq-protein-coupled chemokine receptors upon stimulation with MIP-2 and is crucial for the arrangement of filamentous actin in migrating neutrophils. This is a novel cell function of TRP channel beyond their well-recognized role as universal cell sensors.
Damann
N
N
Department of Molecular Cell Biology, Laboratory of Ion Channel Research, KU Leuven, Leuven, Belgium.
Owsianik
G
G
Li
S
S
Poll
C
C
Nilius
B
B
D000818:Animals; D002118:Calcium / Q000378:metabolism*; D002465:Cell Movement; D054426:Chemokine CXCL2 / Q000235:genetics / Q000378:metabolism*; D005786:Gene Expression Regulation / Q000502:physiology*; D051379:Mice; D018345:Mice, Knockout; D009504:Neutrophils / Q000166:cytology / Q000502:physiology*; D010590:Phalloidine; D011485:Protein Binding; D050052:TRPC Cation Channels / Q000235:genetics / Q000378:metabolism*; D000074287:TRPC6 Cation Channel
D054426:Chemokine CXCL2; D050052:TRPC Cation Channels; D000074287:TRPC6 Cation Channel; C492146:Trpc6 protein, mouse; D010590:Phalloidine; D002118:Calcium