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. 2022 Jan 12;42(2):166–182. doi: 10.1523/JNEUROSCI.2926-20.2021

Figure 5.

Figure 5.

Gephyrin expression is essential for chloride extrusion in hippocampal neurons. A, Representative widefield images of Cl-sensor fluorescence of hippocampal neurons expressing Cl-sensor and either shNT or shGphn, on excitation at 440 and 480 nm. Right, Ratio of the two images at 440 and 480 nm illumination. B, Summary boxplots showing F440/F480 ratios from 17 (shNT) and 14 (shGphn) neurons from two independent experiments. *p < 0.05, Mann–Whitney rank-sum test. C, Left, Representation of experimental setting. RuBi-GABA (30 μm) was locally photolyzed onto the soma or dendrite of transfected neurons whole-cell patch clamped with a high-chloride intracellular solution. Right, Representative recordings of GABA-evoked currents evoked at varying potentials by RuBi-GABA photolysis on the soma or dendrite of hippocampal neurons (DIV 21–24) expressing either shNT or shGphn. D, I–V relations corresponding to traces shown in C. E, Summary graphs showing reversal potential (EGABA, top) and somatodendritic EGABA gradient (ΔEGABA, bottom) in shNT- and shGphn-expressing neurons either in basal culture conditions (n = 9 neurons each, two independent cultures) or on 24 h treatment with TTX and glutamate receptor antagonists (d-APV, NBQX, and MCPG; see Materials and Methods; n = 15 neurons each, n = 2 independent cultures). **p < 0.01, ***p < 0.001; Mann–Whitney rank-sum tests.