Reference - Detail
| RRC ID | 86725 |
|---|---|
| Author | Linsley JW, Shah K, Castello N, Chan M, Haddad D, Doric Z, Wang S, Leks W, Mancini J, Oza V, Javaherian A, Nakamura K, Kokel D, Finkbeiner S. |
| Title | Genetically encoded cell-death indicators (GEDI) to detect an early irreversible commitment to neurodegeneration. |
| Journal | Nat Commun |
| Abstract |
Cell death is a critical process that occurs normally in health and disease. However, its study is limited due to available technologies that only detect very late stages in the process or specific death mechanisms. Here, we report the development of a family of fluorescent biosensors called genetically encoded death indicators (GEDIs). GEDIs specifically detect an intracellular Ca2+ level that cells achieve early in the cell death process and that marks a stage at which cells are irreversibly committed to die. The time-resolved nature of a GEDI delineates a binary demarcation of cell life and death in real time, reformulating the definition of cell death. We demonstrate that GEDIs acutely and accurately report death of rodent and human neurons in vitro, and show that GEDIs enable an automated imaging platform for single cell detection of neuronal death in vivo in zebrafish larvae. With a quantitative pseudo-ratiometric signal, GEDIs facilitate high-throughput analysis of cell death in time-lapse imaging analysis, providing the necessary resolution and scale to identify early factors leading to cell death in studies of neurodegeneration. |
| Volume | 12(1) |
| Pages | 5284 |
| Published | 2021-9-6 |
| DOI | 10.1038/s41467-021-25549-9 |
| PII | 10.1038/s41467-021-25549-9 |
| PMID | 34489414 |
| PMC | PMC8421388 |
| MeSH | Animals Biosensing Techniques* Calcium / metabolism Cell Death / genetics* Cerebral Cortex / cytology Cerebral Cortex / metabolism DNA-Binding Proteins / genetics DNA-Binding Proteins / metabolism Disease Models, Animal Embryo, Nonmammalian Fluorescent Dyes / chemistry Gene Expression Regulation, Developmental* Genes, Reporter Glutamic Acid / pharmacology Green Fluorescent Proteins / genetics Green Fluorescent Proteins / metabolism Humans Larva / cytology Larva / genetics Larva / growth & development Larva / metabolism Mice Mice, Inbred C57BL Neurodegenerative Diseases / genetics* Neurodegenerative Diseases / metabolism Neurodegenerative Diseases / pathology Neurons / cytology Neurons / drug effects Neurons / metabolism* Primary Cell Culture Rats Rats, Long-Evans Single-Cell Analysis / methods Superoxide Dismutase-1 / genetics Superoxide Dismutase-1 / metabolism Zebrafish / embryology Zebrafish / genetics Zebrafish / metabolism alpha-Synuclein / genetics alpha-Synuclein / metabolism |
| IF | 12.121 |
| Resource | |
| Zebrafish | UAS:BoTxBLCGFP, UAS:GCaMP7a |