TY  - JOUR
AU  - Müller, Franziska E
AU  - Cherkas, Volodymyr
AU  - Stopper, Gebhard
AU  - Caudal, Laura C
AU  - Stopper, Laura
AU  - Kirchhoff, Frank
AU  - Henneberger, Christian
AU  - Ponimaskin, Evgeni G
AU  - Zeug, Andre
TI  - Elucidating regulators of astrocytic Ca2+ signaling via multi-threshold event detection (MTED).
JO  - Glia
VL  - 69
IS  - 12
SN  - 1098-1136
CY  - Bognor Regis [u.a.]
PB  - Wiley-Liss
M1  - DZNE-2021-01368
SP  - 2798 - 2811
PY  - 2021
N1  - (CC BY-NC)
AB  - 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.
KW  - Animals
KW  - Astrocytes: metabolism
KW  - Brain: diagnostic imaging
KW  - Brain: metabolism
KW  - Calcium: metabolism
KW  - Calcium Signaling: physiology
KW  - Mice
KW  - Neurons: metabolism
KW  - Ca2+ (Other)
KW  - GCaMP (Other)
KW  - MTED (Other)
KW  - astrocyte (Other)
KW  - biosensor (Other)
LB  - PUB:(DE-HGF)16
C6  - pmid:34388285
DO  - DOI:10.1002/glia.24070
UR  - https://pub.dzne.de/record/162711
ER  -