title: The native TRPP2-dependent channel of murine renal primary cilia.
abstract: Autosomal dominant polycystic kidney disease (ADPKD) is the most common life-threatening monogenic renal disease. ADPKD results from mutations in either of two proteins: polycystin-1 (also known as PC1 or PKD1) or transient receptor potential cation channel, subfamily P, member 2 (TRPP2, also known as polycystin-2, PC2, or PKD2). Each of these proteins is expressed in the primary cilium that extends from many renal epithelial cells. Existing evidence suggests that the cilium can promote renal cystogenesis, while PC1 and TRPP2 counter this cystogenic effect. To better understand the function of TRPP2, we investigated its electrophysiological properties in the native ciliary membrane. We recorded directly from the cilia of mIMCD-3 cells, a murine cell line of renal epithelial origin. In one-third of cilia examined, a large-conductance channel was observed. The channel was not permeable to Cl¯ but conducted cations with permeability ratios PK:PCa:PNaof 1:0.55:0.14. The single-channel conductance ranged from 97 pS in typical physiological solutions to 189 pS in symmetrical 145 mM KCl. Open probability of the channel was very sensitive to membrane depolarization or increasing cytoplasmic free Ca2+in the low micromolar range, with the open probability increasing in either case. Knocking out TRPP2 by CRISPR/Cas9 genome editing eliminated the channel current, establishing it as TRPP2 dependent. Possible mechanisms for activating the TRPP2-dependent channel in the renal primary cilium are discussed.
authors.affiliation: Department of Molecular and Cellular Physiology, University of Cincinnati, Cincinnati, Ohio steve@syrano.acb.uc.edu.
authors.forename: Steven J
authors.identifier: 0000-0001-7663-3590
authors.initials: SJ
authors.lastname: Kleene
authors.affiliation: Department of Molecular and Cellular Physiology, University of Cincinnati, Cincinnati, Ohio.
authors.forename: Nancy K
authors.initials: NK
authors.lastname: Kleene
mesh_terms: D000818:Animals; D002118:Calcium / Q000378:metabolism; D002460:Cell Line; D002923:Cilia / Q000378:metabolism*; D003593:Cytoplasm / Q000378:metabolism; D004847:Epithelial Cells / Q000378:metabolism*; D055786:Gene Knockout Techniques / Q000379:methods; D007668:Kidney / Q000378:metabolism*; D051379:Mice; D016891:Polycystic Kidney, Autosomal Dominant / Q000378:metabolism; D050396:TRPP Cation Channels / Q000235:genetics / Q000378:metabolism*
chemical_list: D002118:Calcium; D050396:TRPP Cation Channels
keywords: PC2; TRPP2; polycystic kidney disease; polycystin; primary cilium