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000162711 037__ $$aDZNE-2021-01368
000162711 041__ $$aEnglish
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000162711 1001_ $$00000-0003-0525-0714$$aMüller, Franziska E$$b0
000162711 245__ $$aElucidating regulators of astrocytic Ca2+ signaling via multi-threshold event detection (MTED).
000162711 260__ $$aBognor Regis [u.a.]$$bWiley-Liss$$c2021
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000162711 520__ $$aRecent 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.
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000162711 650_7 $$2Other$$aCa2+
000162711 650_7 $$2Other$$aGCaMP
000162711 650_7 $$2Other$$aMTED
000162711 650_7 $$2Other$$aastrocyte
000162711 650_7 $$2Other$$abiosensor
000162711 650_2 $$2MeSH$$aAnimals
000162711 650_2 $$2MeSH$$aAstrocytes: metabolism
000162711 650_2 $$2MeSH$$aBrain: diagnostic imaging
000162711 650_2 $$2MeSH$$aBrain: metabolism
000162711 650_2 $$2MeSH$$aCalcium: metabolism
000162711 650_2 $$2MeSH$$aCalcium Signaling: physiology
000162711 650_2 $$2MeSH$$aMice
000162711 650_2 $$2MeSH$$aNeurons: metabolism
000162711 7001_ $$aCherkas, Volodymyr$$b1
000162711 7001_ $$00000-0003-2496-4755$$aStopper, Gebhard$$b2
000162711 7001_ $$00000-0003-2165-281X$$aCaudal, Laura C$$b3
000162711 7001_ $$00000-0002-9648-6222$$aStopper, Laura$$b4
000162711 7001_ $$00000-0002-2324-2761$$aKirchhoff, Frank$$b5
000162711 7001_ $$0P:(DE-2719)2811625$$aHenneberger, Christian$$b6$$udzne
000162711 7001_ $$00000-0002-4570-5130$$aPonimaskin, Evgeni G$$b7
000162711 7001_ $$00000-0001-9858-5841$$aZeug, Andre$$b8
000162711 773__ $$0PERI:(DE-600)1474828-9$$a10.1002/glia.24070$$gVol. 69, no. 12, p. 2798 - 2811$$n12$$p2798 - 2811$$tGlia$$v69$$x1098-1136$$y2021
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000162711 9201_ $$0I:(DE-2719)1013029$$kAG Henneberger$$lRole of astrocytes and extracellular matrix in signal transmission to the brain$$x0
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