title: TRPV4 participates in the establishment of trailing adhesions and directional persistence of migrating cells.
abstract: Calcium signaling participates in different cellular processes leading to cell migration. TRPV4, a non-selective cation channel that responds to mechano-osmotic stimulation and heat, is also involved in cell migration. However, the mechanistic involvement of TRPV4 in cell migration is currently unknown. We now report that expression of the mutant channel TRPV4-(121)AAWAA (lacking the phosphoinositide-binding site (121)KRWRK(125) and the response to physiological stimuli) altered HEK293 cell migration. Altered migration patterns included periods of fast and persistent motion followed by periods of stalling and turning, and the extension of multiple long cellular protrusions. TRPV4-WT overexpressing cells showed almost complete loss of directionality with frequent turns, no progression, and absence of long protrusions. Traction microscopy revealed higher tractions forces in the tail of TRPV4-(121)AAWAA than in TRPV4-WT expressing cells. These results are consistent with a defective and augmented tail retraction in TRPV4-(121)AAWAA- and TRPV4-WT-expressing cells, respectively. The activity of calpain, a protease implicated in focal adhesion (FA) disassembly, was decreased in TRPV4-(121)AAWAA compared with TRPV4-WT-expressing cells. Consistently, larger focal adhesions were seen in TRPV4-(121)AAWAA compared with TRPV4-WT-expressing HEK293 cells, a result that was also reproduced in T47D and U87 cells. Similarly, overexpression of the pore-dead mutant TRPV4-M680D resumed the TRPV4-(121)AAWAA phenotype presenting larger FA. The migratory phenotype obtained in HEK293 cells overexpressing TRPV4-(121)AAWAA was mimicked by knocking-down TRPC1, a cationic channel that participates in cell migration. Together, our results point to the participation of TRPV4 in the dynamics of trailing adhesions, a function that may require the interplay of TRPV4 with other cation channels or proteins present at the FA sites.
authors.affiliation: Laboratory of Molecular Physiology and Channelopathies, Dept. of Experimental and Health Sciences, Universitat Pompeu Fabra, C/ Dr. Aiguader 88, Barcelona, 08003, Spain.
authors.forename: Sanela
authors.initials: S
authors.lastname: Mrkonjić
authors.affiliation: Laboratory of Molecular Physiology and Channelopathies, Dept. of Experimental and Health Sciences, Universitat Pompeu Fabra, C/ Dr. Aiguader 88, Barcelona, 08003, Spain.
authors.forename: Anna
authors.initials: A
authors.lastname: Garcia-Elias
authors.affiliation: Laboratory of Molecular Physiology and Channelopathies, Dept. of Experimental and Health Sciences, Universitat Pompeu Fabra, C/ Dr. Aiguader 88, Barcelona, 08003, Spain.
authors.forename: Carlos
authors.initials: C
authors.lastname: Pardo-Pastor
authors.affiliation: Institute for Bioengineering of Catalonia, Barcelona, Barcelona, Spain.|Facultat de Medicina, Universitat de Barcelona, and Ciber Enfermedades Respiratorias, Barcelona, Barcelona, Spain.
authors.forename: Elsa
authors.initials: E
authors.lastname: Bazellières
authors.affiliation: Institute for Bioengineering of Catalonia, Barcelona, Barcelona, Spain.|Facultat de Medicina, Universitat de Barcelona, and Ciber Enfermedades Respiratorias, Barcelona, Barcelona, Spain.|Institució Catalana de Recerca i Estudis Avançats (ICREA), Barcelona, Barcelona, Spain.
authors.forename: Xavier
authors.initials: X
authors.lastname: Trepat
authors.affiliation: Laboratory of Ion Channels and TRP Research Platform Leuven, KU Leuven, Leuven, Belgium.
authors.forename: Joris
authors.initials: J
authors.lastname: Vriens
authors.affiliation: Laboratory of Ion Channels and TRP Research Platform Leuven, KU Leuven, Leuven, Belgium.
authors.forename: Debapriya
authors.initials: D
authors.lastname: Ghosh
authors.affiliation: Laboratory of Ion Channels and TRP Research Platform Leuven, KU Leuven, Leuven, Belgium.
authors.forename: Thomas
authors.initials: T
authors.lastname: Voets
authors.affiliation: Laboratory of Molecular Physiology and Channelopathies, Dept. of Experimental and Health Sciences, Universitat Pompeu Fabra, C/ Dr. Aiguader 88, Barcelona, 08003, Spain.
authors.forename: Rubén
authors.initials: R
authors.lastname: Vicente
authors.affiliation: Laboratory of Molecular Physiology and Channelopathies, Dept. of Experimental and Health Sciences, Universitat Pompeu Fabra, C/ Dr. Aiguader 88, Barcelona, 08003, Spain. miguel.valverde@upf.edu.
authors.forename: Miguel A
authors.initials: MA
authors.lastname: Valverde
mesh_terms: D001665:Binding Sites; D002154:Calpain / Q000378:metabolism; D002448:Cell Adhesion; D045744:Cell Line, Tumor; D021961:Cell Membrane Structures / Q000378:metabolism*; D002465:Cell Movement; D057809:HEK293 Cells; D006801:Humans; D009154:Mutation; D010716:Phosphatidylinositols / Q000378:metabolism; D011485:Protein Binding; D050916:TRPV Cation Channels / Q000737:chemistry / Q000235:genetics / Q000378:metabolism*
chemical_list: D010716:Phosphatidylinositols; D050916:TRPV Cation Channels; C416986:TRPV4 protein, human; D002154:Calpain
keywords: Calcium; Calpain; Focal adhesion; Migration; TRPV4; Traction forces