Figure 3. Pharmacological treatment with Bumetanide affects GABAergic currents and regulates the expression of α3-α1 and δ subunits.

(a) Example of I–V relations of isoguvacine (10 μM) evoked GABA_A currents at increasing holding potentials in control neurons. (b) Example of I–V relations of isoguvacine (10 μM) evoked GABA_A currents at increasing holding potentials in Bumetanide treated neurons. (c) Summary of E_Cl at DIV6–7 in control (grey bar n=11) and Bumetanide treated neurons (black filled bar n=13; *P<0.05; unpaired t-test). (d) Representative sIPSCs from control and Bumetanide (30 μM) treated neurons. (e) Summary of sIPSC decay kinetics in control (grey bar n=26) and Bumetanide treated neurons (black filled bar n=37; ***P<0.001; unpaired t-test). (f) Summary of sIPSC frequency in control (grey bar n=49) and Bumetanide treated neurons (black filled bar n=45; P>0.05; unpaired t-test). (g) Summary of tonic current amplitude in control (grey bar n=46) and Bumetanide treated neurons (black filled bar n=50; **P<0.01; unpaired t-test). (h) Quantification of immunofluorescence signals for α3 and α1 subunits (α3 control, grey bar n=100; Bumetanide, black filled bar n=99; **P<0.01; unpaired t-test; α1 control, grey bar n=121; Bumetanide, black filled bar n=119; ***P<0.001; unpaired t-test). (i) Quantification of immunofluorescence signals for δ subunit in control (grey bar n=128) and in Bumetanide treated neurons (black filled bar n=96; **P<0.01; unpaired t-test) at DIV6–7. (j) Somatic immunostaining of immunofluorescence signals for α3 and α1 subunits in control and Bumetanide treated neurons at DIV7. (k) Somatic immunostaining of immunofluorescence signals for δ subunit in control and Bumetanide treated neurons at DIV7. Scale bar: 23 μm. Data is presented as mean±s.e.m.