title: Fibroblast growth factor receptor 3 regulates microtubule formation and cell surface mechanical properties in the developing organ of Corti.
abstract: Fibroblast Growth Factor (Fgf) signaling is involved in the exquisite cellular patterning of the developing cochlea, and is necessary for proper hearing function. Our previous data indicate that Fgf signaling disrupts actin, which impacts the surface stiffness of sensory outer hair cells (OHCs) and non-sensory supporting pillar cells (PCs) in the organ of Corti. Here, we used Atomic Force Microscopy (AFM) to measure the impact of loss of function of Fgf-receptor 3, on cytoskeletal formation and cell surface mechanical properties. We find a 50% decrease in both OHC and PC surface stiffness, and a substantial disruption in microtubule formation in PCs. Moreover, we find no change in OHC electromotility of Fgfr3-deficient mice. To further understand the regulation by Fgf-signaling on microtubule formation, we treated wild-type cochlear explants with Fgf-receptor agonist Fgf2, or antagonist SU5402, and find that both treatments lead to a significant reduction in β-Tubulin isotypes I&II. To identify downstream transcriptional targets of Fgf-signaling, we used QPCR arrays to probe 84 cytoskeletal regulators. Of the 5 genes significantly upregulated following treatment, Clasp2, Mapre2 and Mark2 impact microtubule formation. We conclude that microtubule formation is a major downstream effector of Fgf-receptor 3, and suggest this pathway impacts the formation of fluid spaces in the organ of Corti.
authors.affiliation: Auditory Mechanics Section, Laboratory of Cellular Biology, National Institute on Deafness and Other Communication Disorders, National Institutes of Health, Bethesda, MD, USA. szaramak@nidcd.nih.gov
authors.forename: Katherine B
authors.initials: KB
authors.lastname: Szarama
authors.forename: Ruben
authors.initials: R
authors.lastname: Stepanyan
authors.forename: Ronald S
authors.initials: RS
authors.lastname: Petralia
authors.forename: Nuria
authors.initials: N
authors.lastname: Gavara
authors.forename: Gregory I
authors.initials: GI
authors.lastname: Frolenkov
authors.forename: Matthew W
authors.initials: MW
authors.lastname: Kelley
authors.forename: Richard S
authors.initials: RS
authors.lastname: Chadwick
mesh_terms: D000818:Animals; D001696:Biomechanical Phenomena; D002462:Cell Membrane / Q000378:metabolism*; D005346:Fibroblast Growth Factors / Q000378:metabolism; D018072:Hair Cells, Auditory, Outer / Q000378:metabolism / Q000473:pathology / Q000648:ultrastructure; D051379:Mice; D018345:Mice, Knockout; D008870:Microtubules / Q000378:metabolism*; D009925:Organ of Corti / Q000254:growth & development* / Q000378:metabolism* / Q000648:ultrastructure; D051498:Receptor, Fibroblast Growth Factor, Type 3 / Q000378:metabolism*; D015398:Signal Transduction
chemical_list: D005346:Fibroblast Growth Factors; D051498:Receptor, Fibroblast Growth Factor, Type 3