RT Journal Article
SR Electronic
T1 Using SuperClomeleon to Measure Changes in Intracellular Chloride during Development and after Early Life Stress
JF eneuro
JO eNeuro
FD Society for Neuroscience
SP ENEURO.0416-22.2022
DO 10.1523/ENEURO.0416-22.2022
VO 9
IS 6
A1 Lotte J. Herstel
A1 Carlijn Peerboom
A1 Sten Uijtewaal
A1 Dunya Selemangel
A1 Henk Karst
A1 Corette J. Wierenga
YR 2022
UL http://www.eneuro.org/content/9/6/ENEURO.0416-22.2022.abstract
AB Intraneuronal chloride concentrations ([Cl−]i) decrease during development resulting in a shift from depolarizing to hyperpolarizing GABA responses via chloride-permeable GABAA receptors. This GABA shift plays a pivotal role in postnatal brain development, and can be strongly influenced by early life experience. Here, we assessed the applicability of the recently developed fluorescent SuperClomeleon (SClm) sensor to examine changes in [Cl−]i using two-photon microscopy in brain slices. We used SClm mice of both sexes to monitor the developmental decrease in neuronal chloride levels in organotypic hippocampal cultures. We could discern a clear reduction in [Cl−]i between day in vitro (DIV)3 and DIV9 (equivalent to the second postnatal week in vivo) and a further decrease in some cells until DIV22. In addition, we assessed alterations in [Cl−]i in the medial prefrontal cortex (mPFC) of postnatal day (P)9 male SClm mouse pups after early life stress (ELS). ELS was induced by limiting nesting material between P2 and P9. ELS induced a shift toward higher (i.e., immature) chloride levels in layer 2/3 cells in the mPFC. Although conversion from SClm fluorescence to absolute chloride concentrations proved difficult, our study underscores that the SClm sensor is a powerful tool to measure physiological changes in [Cl−]i in brain slices.