Functional TRPV4 channels are expressed in human airway smooth muscle cells.
Hypotonic stimulation induces airway constriction in normal and asthmatic airways. However, the osmolarity sensor in the airway has not been characterized. TRPV4 (also known as VR-OAC, VRL-2, TRP12, OTRPC4), an osmotic-sensitive cation channel in the transient receptor potential (TRP) channel family, was recently cloned. In the present study, we show that TRPV4 mRNA was expressed in cultured human airway smooth muscle cells as analyzed by RT-PCR. Hypotonic stimulation induced Ca(2+) influx in human airway smooth muscle cells in an osmolarity-dependent manner, consistent with the reported biological activity of TRPV4 in transfected cells. In cultured muscle cells, 4alpha-phorbol 12,13-didecanoate (4-alphaPDD), a TRPV4 ligand, increased intracellular Ca(2+) level only when Ca(2+) was present in the extracellular solution. The 4-alphaPDD-induced Ca(2+) response was inhibited by ruthenium red (1 microM), a known TRPV4 inhibitor, but not by capsazepine (1 microM), a TRPV1 antagonist, indicating that 4-alphaPDD-induced Ca(2+) response is mediated by TRPV4. Verapamil (10 microM), an L-type voltage-gated Ca(2+) channel inhibitor, had no effect on the 4-alphaPDD-induced Ca(2+) response, excluding the involvement of L-type Ca(2+) channels. Furthermore, hypotonic stimulation elicited smooth muscle contraction through a mechanism dependent on membrane Ca(2+) channels in both isolated human and guinea pig airways. Hypotonicity-induced airway contraction was not inhibited by the L-type Ca(2+) channel inhibitor nifedipine (1 microM) or by the TRPV1 inhibitor capsazepine (1 microM). We conclude that functional TRPV4 is expressed in human airway smooth muscle cells and may act as an osmolarity sensor in the airway.
Jia
Yanlin
Y
Neurobiology, Schering-Plough Research Institute, Kenilworth, NJ 07033, USA. yanlin.jia@spcorp.com)
Wang
Xin
X
Varty
LoriAnn
L
Rizzo
Charles A
CA
Yang
Richard
R
Correll
Craig C
CC
Phelps
P Tara
PT
Egan
Robert W
RW
Hey
John A
JA
D000818:Animals; D001980:Bronchi / Q000166:cytology*; D002118:Calcium / Q000378:metabolism; D027682:Cation Transport Proteins / Q000235:genetics* / Q000378:metabolism*; D002478:Cells, Cultured; D015870:Gene Expression; D006168:Guinea Pigs; D006801:Humans; D007038:Hypotonic Solutions / Q000494:pharmacology; D007473:Ion Channels / Q000235:genetics* / Q000378:metabolism*; D009119:Muscle Contraction / Q000187:drug effects; D032389:Myocytes, Smooth Muscle / Q000166:cytology / Q000502:physiology*; D050916:TRPV Cation Channels; D014132:Trachea / Q000166:cytology / Q000502:physiology; D014882:Water-Electrolyte Balance / Q000502:physiology
D027682:Cation Transport Proteins; D007038:Hypotonic Solutions; D007473:Ion Channels; D050916:TRPV Cation Channels; C416986:TRPV4 protein, human; D002118:Calcium