title: Flightin is necessary for length determination, structural integrity, and large bending stiffness of insect flight muscle thick filaments.
abstract: Despite the fundamental role of thick filaments in muscle contraction, little is known about the mechanical behavior of these filaments and how myosin-associated proteins dictate differences between muscle types. In this study, we used atomic force microscopy to study the morphological and mechanical properties of fully hydrated native thick filaments isolated from indirect flight muscle (IFM) of normal and mutant Drosophila lacking flightin (fln(0)). IFM thick filaments from newly eclosed (0-1 h old) wild-type flies have a mean length of 3.04+/-0.05 microm. In contrast, IFM thick filaments from newly eclosed fln(0) flies are more variable in length and, on average, are significantly longer (3.90+/-1.33 microm) than wild-type filaments from flies of the same age. In the absence of flightin, thick filaments can attain lengths >300% of wild-type filaments, indicating that flightin is required for setting the proper filament length in vivo. Filaments lacking flightin are structurally compromised, and filament preparations from fully matured 3- to 5-day-old adult fln(0) IFM yielded fragments of variable length much shorter than 3.20+/-0.04 microm, the length obtained from wild-type flies of similar age. The persistence length, an index of bending stiffness, was calculated from measurements of filament end-to-end length and contour length. We show that the presence of flightin increases persistence length by more than 40% and that wild-type filaments increase in stiffness with age. These results indicate that flightin fulfills an essential role in defining the structural and mechanical properties of IFM thick filaments.
authors.affiliation: Department of Biology, University of Vermont, Burlington, VT 05405, USA.
authors.forename: John L
authors.initials: JL
authors.lastname: Contompasis
authors.forename: Lori R
authors.initials: LR
authors.lastname: Nyland
authors.forename: David W
authors.initials: DW
authors.lastname: Maughan
authors.forename: Jim O
authors.initials: JO
authors.lastname: Vigoreaux
mesh_terms: D000818:Animals; D001696:Biomechanical Phenomena; D029721:Drosophila Proteins / Q000172:deficiency / Q000235:genetics / Q000502:physiology*; D004331:Drosophila melanogaster / Q000235:genetics / Q000502:physiology* / Q000648:ultrastructure; D064448:Filamins; D005426:Flight, Animal / Q000502:physiology; D017344:Genes, Insect; D018625:Microscopy, Atomic Force; D020409:Molecular Motor Proteins / Q000172:deficiency / Q000235:genetics / Q000502:physiology; D009124:Muscle Proteins / Q000172:deficiency / Q000235:genetics / Q000502:physiology*; D018482:Muscle, Skeletal / Q000502:physiology* / Q000648:ultrastructure; D009154:Mutation
chemical_list: D029721:Drosophila Proteins; D064448:Filamins; D020409:Molecular Motor Proteins; D009124:Muscle Proteins; C081072:fln protein, Drosophila