TY - JOUR
AU - Groschup, Bernhard
AU - Calandra, Gian-Marco
AU - Raitmayr, Constanze
AU - Shrouder, Joshua
AU - Llovera, Gemma
AU - Zaki, Asal Ghaffari
AU - Burgstaller, Sandra
AU - Bischof, Helmut
AU - Eroglu, Emrah
AU - Liesz, Arthur
AU - Malli, Roland
AU - Filser, Severin
AU - Plesnila, Nikolaus
TI - Probing intracellular potassium dynamics in neurons with the genetically encoded sensor lc-LysM GEPII 1.0 in vitro and in vivo.
JO - Scientific reports
VL - 14
IS - 1
SN - 2045-2322
CY - [London]
PB - Macmillan Publishers Limited, part of Springer Nature
M1 - DZNE-2024-00767
SP - 13753
PY - 2024
AB - Neuronal activity is accompanied by a net outflow of potassium ions (K+) from the intra- to the extracellular space. While extracellular [K+] changes during neuronal activity are well characterized, intracellular dynamics have been less well investigated due to lack of respective probes. In the current study we characterized the FRET-based K+ biosensor lc-LysM GEPII 1.0 for its capacity to measure intracellular [K+] changes in primary cultured neurons and in mouse cortical neurons in vivo. We found that lc-LysM GEPII 1.0 can resolve neuronal [K+] decreases in vitro during seizure-like and intense optogenetically evoked activity. [K+] changes during single action potentials could not be recorded. We confirmed these findings in vivo by expressing lc-LysM GEPII 1.0 in mouse cortical neurons and performing 2-photon fluorescence lifetime imaging. We observed an increase in the fluorescence lifetime of lc-LysM GEPII 1.0 during periinfarct depolarizations, which indicates a decrease in intracellular neuronal [K+]. Our findings suggest that lc-LysM GEPII 1.0 can be used to measure large changes in [K+] in neurons in vitro and in vivo but requires optimization to resolve smaller changes as observed during single action potentials.
KW - Animals
KW - Potassium: metabolism
KW - Neurons: metabolism
KW - Mice
KW - Biosensing Techniques: methods
KW - Action Potentials
KW - Cells, Cultured
KW - Fluorescence Resonance Energy Transfer: methods
KW - Optogenetics: methods
KW - Potassium (NLM Chemicals)
LB - PUB:(DE-HGF)16
C6 - pmid:38877089
C2 - pmc:PMC11178854
DO - DOI:10.1038/s41598-024-62993-1
UR - https://pub.dzne.de/record/270295
ER -