Toward a new insight of calcium oxalate stones in Drosophila by micro-computerized tomography
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We previously developed an animal model of calcium oxalate (CaOx) deposition on the Malphigian tubules of Drosophila melanogaster as a model of urolithiasis. Here, we introduce a new tool for the study of anatomical structure for Drosophila. As a consequence of technical development, the invention of micro-computerized tomography (CT) has been introduced to the small animal, such as rat and mice. We used Drosophila as a model organism and fed the flies 0.5% lithogenic agent ethylene glycol for 3 weeks. Samples were simply prepared for further scanned by micro-CT to scan samples at 800 nm resolution. CT scanning was performed at 40 kVp of voltage, 250 μA of current, and 1750 ms of exposure time and without filter. Reconstruction of sections was carried out with the GPU-based scanner software. Specific region of interests was further analyzed by DataViewer software. Area with high radiologic density level was defined as CaOx deposition for further 3D analysis. Image of whole lithogenic Drosophila was compared with control. High radiologic density level was detected in the region of Malphigian tubules which can be identified as CaOx stones. There was no stone image in the control group. The image was the same as human non-contrast CT for the diagnosis of stone disease. Micro-CT clearly demonstrated the calcium oxalate calcifications in the Malphigian tubules of fruit fly. The image system provides that a new vision on study animal will facilitate further study of stone disease. With the development of new technology on micro-CT, more delicate and advanced image will be presented in the future.
KeywordsCalcium oxalate Drosophila melanogaster Ethylene glycol Micro-computerized tomography Nephrolithiasis/urolithiasis
This study was supported in part by the Taiwan Ministry of Health and Welfare Clinical Trial and Research Center of Excellence (MOHW106-TDU-B-212-113004), Taiwan Ministry of Science and Technology (MOST104-2320-B-039-016-MY3), China Medical University Hospital (DMR-105-048), and CMU under the Aim for Top University Plan of the Taiwan Ministry of Education (A-5-2-A). Y.H.C. and W.Y.L contributed equally to this study. The authors thank Hygene Biotech Co. LTD. for experimental assistance.
WCC and YHC took the primary role in writing the manuscript. WCC designed most of the experiments, with HYC, PCL, SJW, WYL, and YHC, and supervised the final experiments. WCC obtained the funding and conceived, designed, and supervised the study. WCC provided the original ideas for this study.
Compliance with ethical standards
Conflict of interest
All the authors declared no competing interests.
Movie S1. Three-dimensional dynamic micro-CT imaging of the lithogenic Drosophila (MP4 32768 KB)
Movie S2. Three-dimensional dynamic micro-CT imaging of the control Drosophila (MP4 31149 KB)
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