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双色光遗传学工具可使黑腹果蝇心脏停搏、心动过缓和心动过速起搏。

Dual color optogenetic tool enables heart arrest, bradycardic, and tachycardic pacing in Drosophila melanogaster.

机构信息

Department of Biomedical Engineering, Washington University in St Louis, 1 Brookings Dr, St Louis, MO, USA.

Department of Computer Science and Engineering, Washington University in St Louis, 1 Brookings Dr, St Louis, MO, USA.

出版信息

Commun Biol. 2024 Aug 27;7(1):1056. doi: 10.1038/s42003-024-06703-7.

Abstract

In order to facilitate cardiovascular research to develop non-invasive optical heart pacing methods, we have generated a double-transgenic Drosophila melanogaster (fruit fly) model suitable for optogenetic pacing. We created a fly stock with both excitatory H134R-ChR2 and inhibitory eNpHR2.0 opsin transgenes. Opsins were expressed in the fly heart using the Hand-GAL4 driver. Here we describe Hand > H134R-ChR2; eNpHR2.0 model characterization including bi-directional heart control (activation and inhibition) upon illumination of light with distinct wavelengths. Optical control and real-time visualization of the heart function were achieved non-invasively using an integrated light stimulation and optical coherence microscopy (OCM) system. OCM produced high-speed and high-resolution imaging; simultaneously, the heart function was modulated by blue (470 nm) or red (617 nm) light pulses causing tachycardia, bradycardia and restorable cardiac arrest episodes in the same animal. The irradiance power levels and illumination schedules were optimized to achieve successful non-invasive bi-directional heart pacing in Drosophila larvae and pupae.

摘要

为了促进心血管研究以开发非侵入性的光学心脏起搏方法,我们构建了一个适合光遗传学起搏的双转基因黑腹果蝇(果蝇)模型。我们创建了一个带有兴奋性 H134R-ChR2 和抑制性 eNpHR2.0 光敏感蛋白转基因的果蝇品系。通过 Hand-GAL4 驱动子在果蝇心脏中表达这些光敏感蛋白。在这里,我们描述了 Hand > H134R-ChR2; eNpHR2.0 模型的特征,包括在不同波长的光照下进行双向心脏控制(激活和抑制)。使用集成的光刺激和光相干显微镜(OCM)系统,实现了对心脏功能的非侵入性光学控制和实时可视化。OCM 产生高速和高分辨率的图像;同时,通过蓝色(470nm)或红色(617nm)光脉冲调节心脏功能,在同一动物中引起心动过速、心动过缓和可恢复性心脏骤停发作。优化辐照度功率水平和照明方案,以实现在果蝇幼虫和蛹中的成功的非侵入性双向心脏起搏。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/dcd4/11349975/774eaa584e90/42003_2024_6703_Fig1_HTML.jpg

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