Glial Chloride Homeostasis Under Transient Ischemic Stress

Miriam Engels; Manu Kalia; Sarah Rahmati; Laura Petersilie; Peter Kovermann; Michel J A M van Putten; Christine R Rose; Hil G E Meijer; Thomas Gensch; Christoph Fahlke

doi:10.3389/fncel.2021.735300

Glial Chloride Homeostasis Under Transient Ischemic Stress

Front Cell Neurosci. 2021 Sep 16:15:735300. doi: 10.3389/fncel.2021.735300. eCollection 2021.

Authors

Affiliations

¹ Institute of Biological Information Processing, Molekular-und Zellphysiologie (IBI-1), Forschungszentrum Jülich, Jülich, Germany.
² Applied Analysis, Department of Applied Mathematics, University of Twente, Enschede, Netherlands.
³ Institute of Neurobiology, Heinrich Heine University Düsseldorf, Düsseldorf, Germany.
⁴ Department of Clinical Neurophysiology, University of Twente, Enschede, Netherlands.

Abstract

High water permeabilities permit rapid adjustments of glial volume upon changes in external and internal osmolarity, and pathologically altered intracellular chloride concentrations ([Cl^-]_int) and glial cell swelling are often assumed to represent early events in ischemia, infections, or traumatic brain injury. Experimental data for glial [Cl^-]_int are lacking for most brain regions, under normal as well as under pathological conditions. We measured [Cl^-]_int in hippocampal and neocortical astrocytes and in hippocampal radial glia-like (RGL) cells in acute murine brain slices using fluorescence lifetime imaging microscopy with the chloride-sensitive dye MQAE at room temperature. We observed substantial heterogeneity in baseline [Cl^-]_int, ranging from 14.0 ± 2.0 mM in neocortical astrocytes to 28.4 ± 3.0 mM in dentate gyrus astrocytes. Chloride accumulation by the Na⁺-K⁺-2Cl^- cotransporter (NKCC1) and chloride outward transport (efflux) through K⁺-Cl^- cotransporters (KCC1 and KCC3) or excitatory amino acid transporter (EAAT) anion channels control [Cl^-]_int to variable extent in distinct brain regions. In hippocampal astrocytes, blocking NKCC1 decreased [Cl^-]_int, whereas KCC or EAAT anion channel inhibition had little effect. In contrast, neocortical astrocytic or RGL [Cl^-]_int was very sensitive to block of chloride outward transport, but not to NKCC1 inhibition. Mathematical modeling demonstrated that higher numbers of NKCC1 and KCC transporters can account for lower [Cl^-]_int in neocortical than in hippocampal astrocytes. Energy depletion mimicking ischemia for up to 10 min did not result in pronounced changes in [Cl^-]_int in any of the tested glial cell types. However, [Cl^-]_int changes occurred under ischemic conditions after blocking selected anion transporters. We conclude that stimulated chloride accumulation and chloride efflux compensate for each other and prevent glial swelling under transient energy deprivation.

Keywords: K-Cl cotransporters; Na-K-2Cl cotransporter; chemical stress mimicking ischemia; excitatory amino acid transporters; fluorescence lifetime imaging microscopy; intracellular chloride concentrations.