title: Tmod1 and CP49 synergize to control the fiber cell geometry, transparency, and mechanical stiffness of the mouse lens.
abstract: The basis for mammalian lens fiber cell organization, transparency, and biomechanical properties has contributions from two specialized cytoskeletal systems: the spectrin-actin membrane skeleton and beaded filament cytoskeleton. The spectrin-actin membrane skeleton predominantly consists of α₂β₂-spectrin strands interconnecting short, tropomyosin-coated actin filaments, which are stabilized by pointed-end capping by tropomodulin 1 (Tmod1) and structurally disrupted in the absence of Tmod1. The beaded filament cytoskeleton consists of the intermediate filament proteins CP49 and filensin, which require CP49 for assembly and contribute to lens transparency and biomechanics. To assess the simultaneous physiological contributions of these cytoskeletal networks and uncover potential functional synergy between them, we subjected lenses from mice lacking Tmod1, CP49, or both to a battery of structural and physiological assays to analyze fiber cell disorder, light scattering, and compressive biomechanical properties. Findings show that deletion of Tmod1 and/or CP49 increases lens fiber cell disorder and light scattering while impairing compressive load-bearing, with the double mutant exhibiting a distinct phenotype compared to either single mutant. Moreover, Tmod1 is in a protein complex with CP49 and filensin, indicating that the spectrin-actin network and beaded filament cytoskeleton are biochemically linked. These experiments reveal that the spectrin-actin membrane skeleton and beaded filament cytoskeleton establish a novel functional synergy critical for regulating lens fiber cell geometry, transparency, and mechanical stiffness.
authors.affiliation: Department of Cell Biology, The Scripps Research Institute, La Jolla, California, United States of America.
authors.forename: David S
authors.initials: DS
authors.lastname: Gokhin
authors.forename: Roberta B
authors.initials: RB
authors.lastname: Nowak
authors.forename: Nancy E
authors.initials: NE
authors.lastname: Kim
authors.forename: Ernest E
authors.initials: EE
authors.lastname: Arnett
authors.forename: Albert C
authors.initials: AC
authors.lastname: Chen
authors.forename: Robert L
authors.initials: RL
authors.lastname: Sah
authors.forename: John I
authors.initials: JI
authors.lastname: Clark
authors.forename: Velia M
authors.initials: VM
authors.lastname: Fowler
mesh_terms: D000199:Actins / Q000378:metabolism; D000818:Animals; D001696:Biomechanical Phenomena; D048430:Cell Shape; D003599:Cytoskeleton / Q000502:physiology / Q000648:ultrastructure; D005136:Eye Proteins / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D007381:Intermediate Filament Proteins / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D007908:Lens, Crystalline / Q000166:cytology* / Q000502:physiology / Q000648:ultrastructure; D051379:Mice; D008810:Mice, Inbred C57BL; D008815:Mice, Inbred Strains; D016297:Mutagenesis, Site-Directed; D055095:Optics and Photonics; D051348:Tropomodulin / Q000235:genetics / Q000378:metabolism / Q000502:physiology*
chemical_list: D000199:Actins; D005136:Eye Proteins; D007381:Intermediate Filament Proteins; C495831:Tmod1 protein, mouse; D051348:Tropomodulin; C084808:phakinin