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. 2020 Apr 24;11:2014. doi: 10.1038/s41467-020-15778-9

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© The Author(s) 2020

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 4 — a Representative images depicting 2-NBDG filling in coupled astrocyte cell bodies in naïve and stress conditions. Scale bars: 10 µm. b Tau values of filling kinetics showing slower coupling following acute stress. Naïve: n = 6 mice, n = 21 cells; Stress: n = 5 mice, n = 23 cells. Error bars: mean ± s.e.m. Mann–Whitney U test. c Mean trace of coupling in naïve and stress conditions with tau value indicated. d schematic diagram illustrating the placement of patch pipette, extracellular recording electrode, and stimulating electrode. e 2P image depicting electrode placement. Dye in the patch pipette passes between astrocytes through gap-junction channels. Inset, transmitted light image. Scale bar: 100 μm. f A strong linear relationship exists between the slope of the fEPSP and the amplitude of the a-fEPSP (n = 6 mice, n = 8 paired recordings). Error bars: mean ± s.e.m. Pearson’s correlation coefficient, r = 0.99, R2 = 0.97, P = 0.0003. g Example traces depicting changes in both a-fEPSP and fEPSP waveform with increased stimulation voltage. The amplitude of the a-fEPSP is calculated by measuring the difference (mV) between the downward peak and the point 30 ms following this peak. Scale bars for a-fEPSP: 0.5 mV, 10 ms. Scale bars for fEPSP: 0.2 mV, 10 ms. h Maximum intensity z-projection depicting lack of coupling between astrocytes in the presence of 100 μM carbenoxolone. Scale bar: 20 µm. i Normalized a-fEPSP amplitude depicting LTP impairment in the presence of the gap-junction channel blocker carbenoxolone (100 μM; n = 4 mice, n = 5 cells) and D,L-APV (50 μM; n = 4 mice, n = 4 cells). Error bars: mean ± s.e.m. Inset, example traces pre- (black) and post- (gray) LTP induction. Scale bars 0.2 mV, 20 ms. j Scatter dot plot depicting the extent of long-term plasticity 30–40 min following theta burst stimulation. naïve: n = 6 mice, n = 7 cells, n = 4 mice, n = 4 cells; CBX: n = 4 mice, n = 5 cells. Error bars: mean ± s.e.m. One-way ANOVA (k) Illustration. l Normalized a-fEPSP amplitude depicting the effect of patching and filling astrocytes with d-lactate on LTP in naïve animals (black). Naïve+d-lactate: n = 4 mice, n = 7 cells. Error bars: mean ± s.e.m. Inset, example average traces pre- and post-LTP in naïve and naïve+D-lactate conditions. Scale bar 0.5 mV, 5 ms in naïve and 0.25 mV, 5 ms in naïve+d-lactate. m Scatter dot plot indicating the extent of LTP when astrocytes were patched with d-lactate. n = 4 mice, n = 7 cells. Error bars: mean ± s.e.m. One-sample t-test.