title: Differential volume regulation and calcium signaling in two ciliary body cell types is subserved by TRPV4 channels.
abstract: Fluid secretion by the ciliary body plays a critical and irreplaceable function in vertebrate vision by providing nutritive support to the cornea and lens, and by maintaining intraocular pressure. Here, we identify TRPV4 (transient receptor potential vanilloid isoform 4) channels as key osmosensors in nonpigmented epithelial (NPE) cells of the mouse ciliary body. Hypotonic swelling and the selective agonist GSK1016790A (EC50 ∼33 nM) induced sustained transmembrane cation currents and cytosolic [Formula: see text] elevations in dissociated and intact NPE cells. Swelling had no effect on [Formula: see text] levels in pigment epithelial (PE) cells, whereas depolarization evoked [Formula: see text] elevations in both NPE and PE cells. Swelling-evoked [Formula: see text] signals were inhibited by the TRPV4 antagonist HC067047 (IC50 ∼0.9 μM) and were absent in Trpv4(-/-) NPE. In NPE, but not PE, swelling-induced [Formula: see text] signals required phospholipase A2 activation. TRPV4 localization to NPE was confirmed with immunolocalization and excitation mapping approaches, whereas in vivo MRI analysis confirmed TRPV4-mediated signals in the intact mouse ciliary body. Trpv2 and Trpv4 were the most abundant vanilloid transcripts in CB. Overall, our results support a model whereby TRPV4 differentially regulates cell volume, lipid, and calcium signals in NPE and PE cell types and therefore represents a potential target for antiglaucoma medications.
authors.affiliation: Department of Ophthalmology & Visual Sciences, University of Utah School of Medicine, Salt Lake City, UT 84132;
authors.forename: Andrew O
authors.initials: AO
authors.lastname: Jo
authors.affiliation: Department of Ophthalmology & Visual Sciences, University of Utah School of Medicine, Salt Lake City, UT 84132;
authors.forename: Monika
authors.initials: M
authors.lastname: Lakk
authors.affiliation: Department of Ophthalmology & Visual Sciences, University of Utah School of Medicine, Salt Lake City, UT 84132;
authors.forename: Amber M
authors.initials: AM
authors.lastname: Frye
authors.affiliation: Department of Ophthalmology & Visual Sciences, University of Utah School of Medicine, Salt Lake City, UT 84132;
authors.forename: Tam T T
authors.initials: TT
authors.lastname: Phuong
authors.affiliation: Department of Ophthalmology & Visual Sciences, University of Utah School of Medicine, Salt Lake City, UT 84132;
authors.forename: Sarah N
authors.initials: SN
authors.lastname: Redmon
authors.affiliation: Department of Anatomy and Cell Biology, Wayne State University, Detroit, MI 48202;
authors.forename: Robin
authors.initials: R
authors.lastname: Roberts
authors.affiliation: Department of Anatomy and Cell Biology, Wayne State University, Detroit, MI 48202; Department of Ophthalmology, Wayne State University, Detroit, MI 48202;
authors.forename: Bruce A
authors.initials: BA
authors.lastname: Berkowitz
authors.affiliation: Department of Ophthalmology & Visual Sciences, University of Utah School of Medicine, Salt Lake City, UT 84132;
authors.forename: Oleg
authors.initials: O
authors.lastname: Yarishkin
authors.affiliation: Department of Ophthalmology & Visual Sciences, University of Utah School of Medicine, Salt Lake City, UT 84132; Center for Translational Medicine, Moran Eye Institute, University of Utah, Salt Lake City, UT 84132; Department of Neurobiology & Anatomy, University of Utah, Salt Lake City, UT 84132; Department of Bioengineering, University of Utah, Salt Lake City, UT 84132 david.krizaj@hsc.utah.edu.
authors.forename: David
authors.initials: D
authors.lastname: Križaj
mesh_terms: D000818:Animals; D002118:Calcium / Q000378:metabolism; D020013:Calcium Signaling / Q000187:drug effects*; D048429:Cell Size; D002478:Cells, Cultured; D002924:Ciliary Body / Q000502:physiology*; D004789:Enzyme Activation; D004847:Epithelial Cells / Q000378:metabolism*; D004848:Epithelium / Q000378:metabolism*; D005901:Glaucoma / Q000473:pathology*; D007930:Leucine / Q000031:analogs & derivatives / Q000494:pharmacology; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D009997:Osmotic Pressure / Q000502:physiology; D054467:Phospholipases A2 / Q000378:metabolism; D013449:Sulfonamides / Q000494:pharmacology; D050916:TRPV Cation Channels / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism*; D014785:Vision, Ocular / Q000502:physiology*
chemical_list: C530602:N-(1-((4-(2-(((2,4-dichlorophenyl)sulfonyl)amino)-3-hydroxypropanoyl)-1-piperazinyl)carbonyl)-3-methylbutyl)-1-benzothiophene-2-carboxamide; D013449:Sulfonamides; D050916:TRPV Cation Channels; C486576:Trpv4 protein, mouse; D054467:Phospholipases A2; D007930:Leucine; D002118:Calcium
keywords: TRPV4; aqueous humor; ciliary body; glaucoma; intraocular pressure