title: Thin filament length in the cardiac sarcomere varies with sarcomere length but is independent of titin and nebulin.
abstract: Thin filament length (TFL) is an important determinant of the force-sarcomere length (SL) relation of cardiac muscle. However, the various mechanisms that control TFL are not well understood. Here we tested the previously proposed hypothesis that the actin-binding protein nebulin contributes to TFL regulation in the heart by using a cardiac-specific nebulin cKO mouse model (αMHC Cre Neb cKO). Atrial myocytes were studied because nebulin expression has been reported to be most prominent in this cell type. TFL was measured in right and left atrial myocytes using deconvolution optical microscopy and staining for filamentous actin with phalloidin and for the thin filament pointed-end with an antibody to the capping protein Tropomodulin-1 (Tmod1). Results showed that TFLs in Neb cKO and littermate control mice were not different. Thus, deletion of nebulin in the heart does not alter TFL. However, TFL was found to be ~0.05μm longer in the right than in the left atrium and Tmod1 expression was increased in the right atrium. We also tested the hypothesis that the length of titin's spring region is a factor controlling TFL by studying the Rbm20(ΔRRM) mouse which expresses titins that are ~500kDa (heterozygous mice) and ~1000kDa (homozygous mice) longer than in control mice. Results revealed that TFL was not different in Rbm20(ΔRRM) mice. An unexpected finding in all genotypes studied was that TFL increased as sarcomeres were stretched (~0.1μm per 0.35μm of SL increase). This apparent increase in TFL reached a maximum at a SL of ~3.0μm where TFL was ~1.05μm. The SL dependence of TFL was independent of chemical fixation or the presence of cardiac myosin-binding protein C (cMyBP-C). In summary, we found that in cardiac myocytes TFL varies with SL in a manner that is independent of the size of titin or the presence of nebulin.
authors.affiliation: Department of Cellular and Molecular Medicine, Sarver Molecular Cardiovascular Research Program, University of Arizona, Tucson, AZ, USA.
authors.forename: Justin
authors.initials: J
authors.lastname: Kolb
authors.affiliation: Department of Cellular and Molecular Medicine, Sarver Molecular Cardiovascular Research Program, University of Arizona, Tucson, AZ, USA.
authors.forename: Frank
authors.initials: F
authors.lastname: Li
authors.affiliation: Department of Cellular and Molecular Medicine, Sarver Molecular Cardiovascular Research Program, University of Arizona, Tucson, AZ, USA.
authors.forename: Mei
authors.initials: M
authors.lastname: Methawasin
authors.affiliation: Department of Cellular and Molecular Medicine, Sarver Molecular Cardiovascular Research Program, University of Arizona, Tucson, AZ, USA.
authors.forename: Maya
authors.initials: M
authors.lastname: Adler
authors.affiliation: Department of Cellular and Molecular Medicine, Sarver Molecular Cardiovascular Research Program, University of Arizona, Tucson, AZ, USA.
authors.forename: Yael-Natalie
authors.initials: YN
authors.lastname: Escobar
authors.affiliation: Department of Cellular and Molecular Medicine, Sarver Molecular Cardiovascular Research Program, University of Arizona, Tucson, AZ, USA.
authors.forename: Joshua
authors.initials: J
authors.lastname: Nedrud
authors.affiliation: Department of Cellular and Molecular Medicine, Sarver Molecular Cardiovascular Research Program, University of Arizona, Tucson, AZ, USA.
authors.forename: Christopher T
authors.initials: CT
authors.lastname: Pappas
authors.affiliation: Department of Cellular and Molecular Medicine, Sarver Molecular Cardiovascular Research Program, University of Arizona, Tucson, AZ, USA.
authors.forename: Samantha P
authors.initials: SP
authors.lastname: Harris
authors.affiliation: Department of Cellular and Molecular Medicine, Sarver Molecular Cardiovascular Research Program, University of Arizona, Tucson, AZ, USA. Electronic address: granzier@email.arizona.edu.
authors.forename: Henk
authors.initials: H
authors.lastname: Granzier
mesh_terms: D000818:Animals; D064211:Connectin / Q000378:metabolism*; D051379:Mice; D018345:Mice, Knockout; D008840:Microfilament Proteins; D008853:Microscopy; D009124:Muscle Proteins / Q000172:deficiency / Q000235:genetics / Q000378:metabolism*; D032383:Myocytes, Cardiac / Q000378:metabolism*; D009210:Myofibrils; D018995:Myosin Heavy Chains / Q000172:deficiency / Q000235:genetics; D012518:Sarcomeres / Q000502:physiology*
chemical_list: D064211:Connectin; D008840:Microfilament Proteins; D009124:Muscle Proteins; C035620:nebulin; D018995:Myosin Heavy Chains
keywords: Cardiac myocyte; Length regulation; Myofilament; Nebulin; Thin filament; Titin