Using Drosophila to study the structural and signaling functions of filamin in muscles.

利用果蝇研究肌肉中细丝蛋白的结构和信号功能。

• 批准号: RGPIN-2022-02984
• 负责人: GonzálezMorales, Nicanor
• 金额: $ 2.04万
• 依托单位: Dalhousie University
• 依托单位国家: 加拿大
• 项目类别: Discovery Grants Program - Individual
• 财政年份: 2022
• 资助国家: 加拿大
• 起止时间: 2022-01-01 至 2023-12-31
• 项目状态: 已结题
• 来源: https://www.nserc-crsng.gc.ca/ase-oro/Details-Detailles_eng.asp?id=745714
• 关键词: Using; Drosophila; study; structural; signaling

The overall objective of this NSERC grant is to understand the mechanisms that mediate muscle contraction. Muscles have an impressive actin cytoskeleton, composed of thousands of repeated contractile units called sarcomeres that allow them to produce large contractile forces. Sarcomeres are composed of antiparallel actin and myosin filaments that slide along each other. We want to know the mechanisms that prevent sarcomeres from breaking apart during muscle contraction. We will focus on the actin-binding protein filamin because of three important observations. It has an elastic region that coils around itself and adopts either a coiled or a linear conformation upon pulling forces. It localizes between two opposing sarcomeres at the perfect spot to resist contractile forces. Most importantly, reducing filamin levels in Drosophila muscles cause the sarcomeres to break. We will use Drosophila to analyze in detail the function of filamin in maintaining sarcomere stability. First, we will use a recently generated scarless genetic engineering method to make mutant flies with specific mutations in the filamin gene. Then, we will analyze the muscles of these mutant flies using confocal, super-resolution and electron microscopy techniques. This combination will allow us to separate the elastic, the structural, and the signalling functions of filamin in muscles. Our preliminary results include the generation of a few mutant fly lines, which demonstrate the feasibility of the genetic manipulation method, and preliminary microscopy images of two filamin mutants in which the elastic property is compromised.

这项NSERC资助的总体目标是了解介导肌肉收缩的机制。肌肉有一个令人印象深刻的肌动蛋白细胞骨架，由数千个重复收缩的单位组成，称为肌节，使它们能够产生巨大的收缩力。肌节由反向平行的肌动蛋白和肌球蛋白细丝组成，它们相互沿着滑动。我们想知道防止肌节在肌肉收缩时破裂的机制。 我们将集中在肌动蛋白结合蛋白细丝蛋白，因为三个重要的观察。它有一个弹性区域，该区域围绕自身盘绕，并且在拉力下采用盘绕或线性构造。它位于两个相对的肌节之间的完美位置以抵抗收缩力。最重要的是，降低果蝇肌肉中的细丝蛋白水平会导致肌节断裂。 我们将使用果蝇来详细分析细丝蛋白在维持肌节稳定性中的功能。首先，我们将使用最近产生的无疤痕基因工程方法，使突变苍蝇与特定的细丝蛋白基因突变。然后，我们将使用共聚焦，超分辨率和电子显微镜技术分析这些突变苍蝇的肌肉。这种结合将使我们能够分离肌肉中细丝蛋白的弹性，结构和信号功能。我们的初步结果包括产生几个突变飞线，这表明遗传操作方法的可行性，和初步的显微镜图像的两个细丝蛋白突变体中的弹性性能受到损害。