High potassium exposure reveals the altered ability of astrocytes to regulate their volume in the aged hippocampus of GFAP/EGFP mice. D. Kolenicova, J. Tureckova, B. Pukajova, L. Harantova, J. Kriska, D. Kirdajova, I. Vorisek, M. Kamenicka, L. Valihrach, P. Androvic, M. Kubista, L. Vargova, M. Anderova. Neurobiology of Aging, 2019, In Press, Corrected Proof. https://doi.org/10.1016/j.neurobiolaging.2019.10.009.
In this study, we focused on age-related changes in astrocyte functioning, predominantly on the ability of astrocytes to regulate their volume in response to a pathological stimulus, namely extracellular 50 mM K+ concentration. The aim of our project was to identify changes in the expression and function of transport proteins in the astrocytic membrane and properties of the extracellular space (ECS), triggered by aging. We employed three-dimensional (3D) confocal morphometry, gene expression profiling, immunohistochemical analysis and diffusion measurement in the hippocampal slices from 3-month- (3M), 9-month- (9M), 12-month- (12M) and 18-month- (18M) old GFAP/EGFP mice, in which astrocytes are visualized by enhanced green fluorescent protein (EGFP) under the control of the promotor for human glial fibrillary acidic protein (GFAP). Combining a pharmacological approach and the quantification of astrocyte volume changes evoked by hyperkalemia, we found that marked diversity in the extent of astrocyte swelling in the hippocampus during aging is due to the gradually declining participation of Na+-K+-Cl- transporters, glutamate transporters (GLAST and GLT-1), and volume-regulated anion channels. Interestingly, there was a redistribution of Na+-K+-Cl- co-transporter, and glutamate transporters from astrocytic soma to processes. In addition, immunohistochemical analysis confirmed an age-dependent decrease in the content of NKCC1 in astrocytes. The overall extracellular volume changes revealed a similar age-dependent diversity during hyperkalemia as observed in astrocytes. In addition, the recovery of the ECS was markedly impaired in aged animals.
Age-dependent changes in K+ uptake, glutamate uptake and astrocyte volume regulation. (A) Stoichiometry of Na+-K+-Cl- cotransporter (NKCC1), glutamate and glutamate-aspartate transporter (GLT-1 and GLAST) and volume-regulated anion channels (VRACs). (B) Schematic diagram depicting changes in the functioning of NKCC1, glutamate transporters and VRACs during aging. In hippocampal astrocytes of 9M-old mice NKCC1 and glutamate transporters contribute mainly to the swelling of astrocytic soma, while VRACs participate in volume regulation of astrocytic processes. In hippocampal astrocytes of 12M- and 18M-old mice the functioning of NKCC1, glutamate transporters and VRACs markedly declines and, moreover, NKCC1 partly contributes to the swelling of astrocytic processes.