Elucidating regulators of astrocytic Ca 2+ signaling via multi‐threshold event detection ( MTED )
| Autor: | Christian Henneberger, Laura Stopper, Franziska E. Müller, Laura C. Caudal, Evgeni Ponimaskin, Frank Kirchhoff, Andre Zeug, Gebhard Stopper, Volodymyr Cherkas |
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| Rok vydání: | 2021 |
| Předmět: |
GCaMP
Biology biosensor Mice 03 medical and health sciences Cellular and Molecular Neuroscience astrocyte 0302 clinical medicine Animals Premovement neuronal activity metabolism [Calcium] ddc:610 diagnostic imaging [Brain] 030304 developmental biology Research data Brain network 0303 health sciences metabolism [Astrocytes] Event (computing) Multidimensional data Functional imaging Ca2+ Neurology metabolism [Brain] metabolism [Neurons] physiology [Calcium Signaling] MTED Neuroscience 030217 neurology & neurosurgery Ca2 signaling |
| Zdroj: | Glia Glia 69(12), 2798-2811 (2021). doi:10.1002/glia.24070 |
| ISSN: | 0894-1491 |
| DOI: | 10.1002/glia.24070 |
| Popis: | Recent achievements in indicator optimization and imaging techniques promote the advancement of functional imaging to decipher complex signaling processes in living cells, such as Ca2+ activity patterns. Astrocytes are important regulators of the brain network and well known for their highly complex morphology and spontaneous Ca2+ activity. However, the astrocyte community is lacking standardized methods to analyze and interpret Ca2+ activity recordings, hindering global comparisons. Here, we present a biophysically-based analytical concept for deciphering the complex spatio-temporal changes of Ca2+ biosensor fluorescence for understanding the underlying signaling mechanisms. We developed a pixel-based multi-threshold event detection (MTED) analysis of multidimensional data, which accounts for signal strength as an additional signaling dimension and provides the experimenter with a comprehensive toolbox for a differentiated and in-depth characterization of fluorescence signals. MTED was validated by analyzing astrocytic Ca2+ activity across Ca2+ indicators, imaging setups, and model systems from primary cell culture to awake, head-fixed mice. We identified extended Ca2+ activity at 25°C compared to 37°C physiological body temperature and dissected how neuronal activity shapes long-lasting astrocytic Ca2+ activity. Our MTED strategy, as a parameter-free approach, is easily transferrable to other fluorescent indicators and biosensors and embraces the additional dimensionality of signaling activity strength. It will also advance the definition of standardized procedures and parameters to improve comparability of research data and reports. |
| Databáze: | OpenAIRE |
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