Using en-face optical coherence tomography to analyse gene function in Drosophila Melanogaster larval heart

A. Bradu, Lisha Ma, J. Bloor, A. Podoleanu
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引用次数: 1

Abstract

In-vivo Optical Coherence Tomography (OCT) imaging of the fruit fly Drosophila Melanogaster larval heart allows non invasive visualizations and assessment of its cardiac function. In order to image Drosophila heart, we have developed a dedicated imaging instrument able to provide simultaneous OCT and Laser Scanning Confocal Microscopy (LSCM) images. With this dual imaging system, the heart can easily be located and visualised within the specimen and the change of the heart shape in a cardiac cycle monitored. Here we have used targeted gene expression to knockdown the myospheroid (mys) gene in the larval heart using a specific RNAi construct. By knocking down a β integrin subunit encoded by mys we have recorded an enlarged heart chamber in both diastolic and systolic states. Also, the fraction of reduction of the chamber diameter was smaller in the knockdown heart. These phenotypic differences indicate that impaired cardiac contractility occurs in the heart where the integrin gene express level is reduced. At our knowledge, this is for the first time when it is shown that integrins have a direct relationship to a dilated heart defect.
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采用正面光学相干断层扫描分析黑腹果蝇幼体心脏基因功能
果蝇黑腹果蝇幼虫心脏的体内光学相干断层扫描(OCT)成像允许无创可视化和评估其心脏功能。为了对果蝇心脏进行成像,我们开发了一种能够同时提供OCT和激光扫描共聚焦显微镜(LSCM)图像的专用成像仪器。使用这种双成像系统,心脏可以很容易地定位和可视化标本内的心脏,并监测心脏周期中心脏形状的变化。在这里,我们使用特定的RNAi构建物,使用靶向基因表达来敲低幼虫心脏中的肌球蛋白(mys)基因。通过敲除mys编码的β整合素亚基,我们记录了舒张和收缩状态下增大的心室。此外,心室直径缩小的比例在打倒心脏更小。这些表型差异表明,心脏收缩能力受损发生在整合素基因表达水平降低的心脏。据我们所知,这是第一次证明整合素与扩张性心脏缺陷有直接关系。
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