title: Heteromultimeric TRPC6-TRPC7 channels contribute to arginine vasopressin-induced cation current of A7r5 vascular smooth muscle cells.
abstract: The molecular identity of receptor-operated, nonselective cation channels (ROCs) of vascular smooth muscle (VSM) cells is not known for certain. Mammalian homologues of the Drosophila canonical transient receptor potential channels (TRPCs) are possible candidates. This study tested the hypothesis that heteromultimeric TRPC channels contribute to ROC current of A7r5 VSM cells activated by [Arg(8)]-vasopressin. A7r5 cells expressed transcripts encoding TRPC1, TRPC4beta, TRPC6, and TRPC7. TRPC4, TRPC6, and TRPC7 protein expression was confirmed by immunoblotting and association of TRPC6 with TRPC7, but not TRPC4beta, was detected by coimmunoprecipitation. The amplitude of arginine vasopressin (AVP)-induced ROC current was suppressed by dominant-negative mutant TRPC6 (TRPC6(DN)) but not TRPC5 (TRPC5(DN)) mutant subunit expression. These data indicate a role for TRPC6- and/or TRPC7-containing channels and rule a more complex subunit composition including TRPC1 and TRPC4. Increasing extracellular Ca(2+) concentration ([Ca(2+)](o)) from 0.05 to 1 mmol/L suppressed currents owing to native, TRPC7, and heteromultimeric TRPC6-TRPC7 channels, but not TRPC6 current, which was slightly enhanced. The relative changes in native and heteromultimeric TRPC6-TRPC7 current amplitudes for [Ca(2+)](o) between approximately 0.01 and 1 mmol/L were identical, but the changes in homomultimeric TRPC6 and TRPC7 currents were significantly less and greater, respectively, compared with the native channels. Taken together, the data provide biochemical and functional evidence supporting the view that heteromultimeric TRPC6-TRPC7 channels contribute to receptor-activated, nonselective cation channels of A7r5 VSM cells.
authors.affiliation: Smooth Muscle Research Group, Faculty of Medicine, University of Calgary, Alberta, Canada.
authors.forename: Yoshiaki
authors.initials: Y
authors.lastname: Maruyama
authors.forename: Yuko
authors.initials: Y
authors.lastname: Nakanishi
authors.forename: Emma J
authors.initials: EJ
authors.lastname: Walsh
authors.forename: David P
authors.initials: DP
authors.lastname: Wilson
authors.forename: Donald G
authors.initials: DG
authors.lastname: Welsh
authors.forename: William C
authors.initials: WC
authors.lastname: Cole
mesh_terms: D001127:Arginine Vasopressin / Q000494:pharmacology*; D002118:Calcium / Q000378:metabolism; D002412:Cations / Q000378:metabolism*; D002460:Cell Line; D045604:Extracellular Fluid / Q000378:metabolism; D005799:Genes, Dominant; D006801:Humans; D007473:Ion Channels / Q000187:drug effects / Q000502:physiology*; D009131:Muscle, Smooth, Vascular / Q000378:metabolism*; D009154:Mutation; D032389:Myocytes, Smooth Muscle / Q000378:metabolism*; D009994:Osmolar Concentration; D011499:Protein Processing, Post-Translational; D012333:RNA, Messenger / Q000378:metabolism; D011994:Recombinant Proteins / Q000378:metabolism; D050052:TRPC Cation Channels / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D000074287:TRPC6 Cation Channel
chemical_list: D002412:Cations; D007473:Ion Channels; D012333:RNA, Messenger; D011994:Recombinant Proteins; D050052:TRPC Cation Channels; D000074287:TRPC6 Cation Channel; C117032:TRPC6 protein, human; C498501:TRPC7 protein, human; D001127:Arginine Vasopressin; D002118:Calcium