Modulation der zytosolischen Redox-Balance durch aktivitätsinduzierte Ca2+-Signale in hippokampalen Neuronen der Maus

Modulation of cytosolic redox balance by activity-induced Ca2+ signals in mouse hippocampal neurons

by Nina Woeste née Böhme
Doctoral thesis
Date of Examination:2025-07-08
Date of issue:2025-06-24
Advisor:Prof. Dr. Michael Müller
Referee:Prof. Dr. Michael Müller
Referee:Prof. Dr. Ivan Bogeski
Referee:Prof. Dr. Dr. Philipp Kauffmann
Persistent Address: http://dx.doi.org/10.53846/goediss-11340

 

 

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Abstract

English

Intracellular Ca2+ signals are essential for the modulation of cellular activity and excitability but also represent critical components of cellular dysfunction and damage in various neurological disorders. The aim of this work is to analyse the interaction of intracellular Ca2+ signals and cytosolic redox balance in mouse hippocampal neurons. Therefore ratiometric fluorescence imaging was used. Measurements based on primary cell cultures showed that there was always a redox balance shift after supplying reagents that increase the intracellular Ca2+ levels, usually toward more oxidized conditions in neurons. Glutamate and in particular the NMDA receptor play a decisive role in this Ca2+ dependent modulation of the redox balance. The technical challenge of this work was the simultaneous quantitative measurement of both signals, cytosolic redox balance and intracellular Ca2+ levels. This was achieved by using the optical redox indicator roGFP1 and the Ca2+ dye Fura-Red. Based on their spectral properties, they can be excited at same wavelengths and the signals can be separated on the emission side by an optical image splitter (Dual View).
Keywords: Calcium signaling; redox balance; reactive oxygen species (ROS); Glutamate; optical imaging; roGFP; Dual View