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. Author manuscript; available in PMC: 2015 Feb 7.
Published in final edited form as: Science. 2014 Feb 7;343(6171):670–675. doi: 10.1126/science.1245423

Fig. 1. [Cl−]i is not solely determined by transport activity.

Fig. 1

A: Two-photon image of Clomeleon YFP from acute hippocampal slice CA1 (left) and neocortex layer IV/V (right). Lower panel: Detailed image of neuron outlined in right panel. B: [P8-9 hippocampal slice] Left, initial vs. new steady state [Cl−]i after 30 min NKCC1 block followed by additional 30 min block of KCC2. Lines, linear fit. Right, as left but KCC2 blocked first then KCC2/NKCC1 block. C: As B but in P32-44 CA1 neurons. D: Transporter index histogram of all neonatal hippocampal neurons (calculated as in methods and Fig. S5), 2.5 ± 5.9 (n = 237; median ± SD). Black circle indicates expected value for canonical co-operation of KCC2 and NKCC1. E: Transporter index of all recorded neonatal CA1 neurons (2.5 ± 5.9), and adult CA1 neurons (1.07 ± 5.6; n = 227) and expected values (neonatal:16.7 ± 6.5; n = 237; adult:15.7 ± 8.9; n = 227). *, P < 0.001, Mann-Whitney Rank Sum Test. F: Upper, [Cl−]i from P6-8 layer IV/V neocortical pyramidal cells (Soma: 16.0 ± 8.9 mM; Dendrite: 11.5 ± 8.3 mM; Axon: 13.9 ± 8.6; P < 0.001, Friedman test, *, P < 0.05 multiple comparison Dunn’s method). Lower, [Cl−]i from P28-30 layer IV/V neocortical pyramidal cells (Soma: 9.3 ± 6.1 mM; Dendrite: 11.4 ± 6.5 mM; Axon: 16.1 ± 7.9 mM; P < 0.001). G: Subcellular gradient differences between axon-soma (A-S) and dendrite-soma (D-S). Blockade of NKCC1 in neonates (10 µM bumetanide) and NKCC1/KCC2 in adults (100 µM bumetanide) did not alter the subcellular [Cl−]i gradients (paired neuron-process; P > 0.05; paired t-test, error lines = SD).