title: The roles of troponin C isoforms in the mechanical function of Drosophila indirect flight muscle.
abstract: Stretch activation (SA) is a fundamental property of all muscle types that increases power output and efficiency, yet its mechanism is unknown. Recently, studies have implicated troponin isoforms as important in the SA mechanism. The highly stretch-activated Drosophila IFMs express two isoforms of the Ca(2+)-binding subunit of troponin (TnC). TnC1 (TnC-F2 in Lethocerus IFM) has two calcium binding sites, while an unusual isoform, TnC4 (TnC-F1 in Lethocerus IFM), has only one binding site. We investigated the roles of these two TnC isoforms in Drosophila IFM by targeting RNAi to each isoform. IFMs with TnC4 expression (normally ~90% of total TnC) replaced by TnC1 did not generate isometric tension, power or display SA. However, TnC4 knockdown resulted in sarcomere ultrastructure disarray, which could explain the lack of mechanical function and thus make interpretation of the influence of TnC4 on SA difficult. Elimination of TnC1 expression (normally ~10% of total TnC) by RNAi resulted in normal muscle structure. In these IFMs, fiber power generation, isometric tension, stretch-activated force and calcium sensitivity were statistically identical to wild type. When TnC1 RNAi was driven by an IFM specific driver, there was no decrease in flight ability or wing beat frequency, which supports our mechanical findings suggesting that TnC1 is not essential for the mechanical function of Drosophila IFM. This finding contrasts with previous work in Lethocerus IFM showing TnC1 is essential for maximum isometric force generation. We propose that differences in TnC1 function in Lethocerus and Drosophila contribute to the ~40-fold difference in IFM isometric tension generated between these species.
authors.affiliation: Department of Biology & Center for Biotechnology and Interdisciplinary Studies, Rensselaer Polytechnic Institute, Troy, NY, 12180, USA.
authors.forename: Catherine C
authors.initials: CC
authors.lastname: Eldred
authors.forename: Anja
authors.initials: A
authors.lastname: Katzemich
authors.forename: Monica
authors.initials: M
authors.lastname: Patel
authors.forename: Belinda
authors.initials: B
authors.lastname: Bullard
authors.forename: Douglas M
authors.initials: DM
authors.lastname: Swank
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified; D004330:Drosophila; D029721:Drosophila Proteins / Q000502:physiology*; D005426:Flight, Animal / Q000502:physiology*; D009119:Muscle Contraction / Q000502:physiology*; D020033:Protein Isoforms / Q000502:physiology; D019209:Troponin C / Q000502:physiology*
chemical_list: D029721:Drosophila Proteins; D020033:Protein Isoforms; D019209:Troponin C