Protocol for a two-clone system to study signaling interactions among neuronal cells in a pre-clinical Alzheimer’s disease model
Summary: This protocol outlines a two-clone genetic mosaic approach in Drosophila to study cell signaling using the FLP/FRT (Flippase/Flippase recombination target) and Gal4/UAS (upstream activating sequence)/Gal80 systems. We generated a stable transgenic line expressing human Aβ42, and then we cro...
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| Format: | Article |
| Language: | English |
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Elsevier
2025-09-01
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| Series: | STAR Protocols |
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| Online Access: | http://www.sciencedirect.com/science/article/pii/S2666166725003995 |
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| author | Aditi Singh Prajakta Deshpande Ankita Sarkar Catherine Yeates Anuradha Venkatakrishnan Chimata Madhuri Kango-Singh Amit Singh |
| author_facet | Aditi Singh Prajakta Deshpande Ankita Sarkar Catherine Yeates Anuradha Venkatakrishnan Chimata Madhuri Kango-Singh Amit Singh |
| author_sort | Aditi Singh |
| collection | DOAJ |
| description | Summary: This protocol outlines a two-clone genetic mosaic approach in Drosophila to study cell signaling using the FLP/FRT (Flippase/Flippase recombination target) and Gal4/UAS (upstream activating sequence)/Gal80 systems. We generated a stable transgenic line expressing human Aβ42, and then we crossed it with a GFP-marked FRT line to produce mosaic clones via heat-shock-induced recombination. Then we dissected, stained, and imaged the eye discs to visualize wild-type and mutant clones. Image analysis with Fiji/ImageJ and statistical quantification in GraphPad Prism enable the study of cell competition and intercellular signaling mechanism(s) in human diseases.For complete details on the use and execution of this protocol, please refer to Yeates et al. and Tare et al.1,2 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. |
| format | Article |
| id | doaj-art-4e9c8588ea7f457799860bb67e6f7449 |
| institution | Kabale University |
| issn | 2666-1667 |
| language | English |
| publishDate | 2025-09-01 |
| publisher | Elsevier |
| record_format | Article |
| series | STAR Protocols |
| spelling | doaj-art-4e9c8588ea7f457799860bb67e6f74492025-08-20T03:59:37ZengElsevierSTAR Protocols2666-16672025-09-016310399310.1016/j.xpro.2025.103993Protocol for a two-clone system to study signaling interactions among neuronal cells in a pre-clinical Alzheimer’s disease modelAditi Singh0Prajakta Deshpande1Ankita Sarkar2Catherine Yeates3Anuradha Venkatakrishnan Chimata4Madhuri Kango-Singh5Amit Singh6Department of Biology, University of Dayton, Dayton, OH 45469, USADepartment of Biology, University of Dayton, Dayton, OH 45469, USADepartment of Biology, University of Dayton, Dayton, OH 45469, USADepartment of Biology, University of Dayton, Dayton, OH 45469, USADepartment of Biology, University of Dayton, Dayton, OH 45469, USADepartment of Biology, University of Dayton, Dayton, OH 45469, USA; Premedical Program, University of Dayton, Dayton, OH, USA; Integrative Science and Engineering (ISE), University of Dayton, Dayton, OH, USA; Center for Genomic Advocacy (TCGA), Indiana State University, Terre Haute, IN, USADepartment of Biology, University of Dayton, Dayton, OH 45469, USA; Premedical Program, University of Dayton, Dayton, OH, USA; Integrative Science and Engineering (ISE), University of Dayton, Dayton, OH, USA; Corresponding authorSummary: This protocol outlines a two-clone genetic mosaic approach in Drosophila to study cell signaling using the FLP/FRT (Flippase/Flippase recombination target) and Gal4/UAS (upstream activating sequence)/Gal80 systems. We generated a stable transgenic line expressing human Aβ42, and then we crossed it with a GFP-marked FRT line to produce mosaic clones via heat-shock-induced recombination. Then we dissected, stained, and imaged the eye discs to visualize wild-type and mutant clones. Image analysis with Fiji/ImageJ and statistical quantification in GraphPad Prism enable the study of cell competition and intercellular signaling mechanism(s) in human diseases.For complete details on the use and execution of this protocol, please refer to Yeates et al. and Tare et al.1,2 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.http://www.sciencedirect.com/science/article/pii/S2666166725003995Cell BiologyDevelopmental biologyGeneticsMicroscopy |
| spellingShingle | Aditi Singh Prajakta Deshpande Ankita Sarkar Catherine Yeates Anuradha Venkatakrishnan Chimata Madhuri Kango-Singh Amit Singh Protocol for a two-clone system to study signaling interactions among neuronal cells in a pre-clinical Alzheimer’s disease model STAR Protocols Cell Biology Developmental biology Genetics Microscopy |
| title | Protocol for a two-clone system to study signaling interactions among neuronal cells in a pre-clinical Alzheimer’s disease model |
| title_full | Protocol for a two-clone system to study signaling interactions among neuronal cells in a pre-clinical Alzheimer’s disease model |
| title_fullStr | Protocol for a two-clone system to study signaling interactions among neuronal cells in a pre-clinical Alzheimer’s disease model |
| title_full_unstemmed | Protocol for a two-clone system to study signaling interactions among neuronal cells in a pre-clinical Alzheimer’s disease model |
| title_short | Protocol for a two-clone system to study signaling interactions among neuronal cells in a pre-clinical Alzheimer’s disease model |
| title_sort | protocol for a two clone system to study signaling interactions among neuronal cells in a pre clinical alzheimer s disease model |
| topic | Cell Biology Developmental biology Genetics Microscopy |
| url | http://www.sciencedirect.com/science/article/pii/S2666166725003995 |
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