Journal Article

DZNE-2024-00767

http://join2-wiki.gsi.de/foswiki/pub/Main/Artwork/join2_logo100x88.png Probing intracellular potassium dynamics in neurons with the genetically encoded sensor lc-LysM GEPII 1.0 in vitro and in vivo.

Groschup, B. ; Calandra, G.-M.Extern* ; Raitmayr, C. ; Shrouder, J. ; Llovera, G. ; Zaki, A. G. ; Burgstaller, S. ; Bischof, H. ; Eroglu, E. ; Liesz, A. ; Malli, R. ; Filser, S. (Last author)DZNE* ; Plesnila, N.

2024
Macmillan Publishers Limited, part of Springer Nature [London]

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Please use a persistent id in citations: doi:10.1038/s41598-024-62993-1

Abstract: 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.

Keyword(s): Animals (MeSH) ; Potassium: metabolism (MeSH) ; Neurons: metabolism (MeSH) ; Mice (MeSH) ; Biosensing Techniques: methods (MeSH) ; Action Potentials (MeSH) ; Cells, Cultured (MeSH) ; Fluorescence Resonance Energy Transfer: methods (MeSH) ; Optogenetics: methods (MeSH) ; Potassium

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Contributing Institute(s):

1. Light Microscopy Facility (CRFS-LMF) (LMF)

Research Program(s):

1. 899 - ohne Topic (POF4-899) (POF4-899)

Experiment(s):

1. Light Microscope Facility (CRFS-LMF) / Bonn

Appears in the scientific report 2024

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