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. 2012 Aug 22;32(34):11716–11726. doi: 10.1523/JNEUROSCI.1942-12.2012

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Copyright © 2012 the authors 0270-6474/12/3211716-11$15.00/0

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Figure 9. — BRAG1 signals synaptic depression via JNK. A, Evoked AMPA-R-mediated (−60 mV) and NMDA-R-mediated (+40 mV) responses recorded from neighboring nonexpressing (Ctrl) and mCherry-tagged BRAG1-IQ or BRAG1-ΔN expressing neurons cultured in media containing 5 μm SP600125 or 2 μm SB203580. B, Values for AMPA responses in neurons expressing BRAG1-IQ and BRAG1-ΔN cultured in media containing SP600125 (Ctrl: −23.0 ± 2.1 pA; IQ: minus]23.5 ± 2.8 pA; n = 26, p = 0.81; Ctrl: −18.4 ± 1.6 pA; ΔN: −19.7 ± 2.0 pA; n = 23, p = 0.48) or SB203580 (Ctrl: −26.0 ± 2.2 pA; IQ: −18.2 ± 1.4 pA; n = 28, p < 0.005; Ctrl: −17.9 ± 1.6 pA; ΔN: −25.5 ± 1.9 pA; n = 23, p < 0.01), and for NMDA responses in neurons expressing BRAG1-IQ and BRAG1-ΔN cultured in media containing SP600125 (Ctrl: 55.4 ± 4.7 pA; IQ: 55.4 ± 6.0 pA; n = 26, p = 0.53; Ctrl: 60.7 ± 10.6 pA; ΔN: 58.3 ± 12.1 pA; n = 23, p = 0.38) or SB203580 (Ctrl: 51.3 ± 6.7 pA; IQ: 54.0 ± 6.2 pA; n = 28, p = 0.93; Ctrl: 60.6 ± 6.3 pA; IQ: 56.4 ± 7.7 pA; n = 23, p = 0.43) related to control nonexpressing neurons. C, Western blots of phospho-p46/54 JNK or p38 MAPK in control hippocampal CA1 regions, hippocampal CA1 regions expressing BRAG1-IQ, and hippocampal CA1 regions expressing BRAG1-ΔN. Each lane was loaded with the same amount of protein (45 μg). D, Left, Relative amounts of phospho-p46 (Ctrl: 100.0 ± 5.3%; IQ: 118.9 ± 7.5%; n = 12; p < 0.005; ΔN: 89.6 ± 5.8%; n = 12; p < 0.01) and phospho-p54 (Ctrl: 100.0 ± 11.0%; IQ: 117.4 ± 11.6%; n = 12; p < 0.005; ΔN: 81.6 ± 9.9%; n = 12; p < 0.005) JNK in hippocampal CA1 regions expressing BRAG1-IQ and BRAG1-ΔN. Relative amounts of total p46 (Ctrl: 100.0 ± 7.7%; IQ: 101.2 ± 8.2%; n = 12; p = 0.64; ΔN: 98.3 ± 8.1%; n = 12; p = 0.43) and total p54 (Ctrl: 100.0 ± 10.2%; IQ: 107.2 ± 11.0%; n = 12; p = 0.14; ΔN: 101.3 ± 10.2%; n = 12; p = 0.24) JNK in hippocampal CA1 regions expressing BRAG1-IQ and BRAG1-ΔN. Right, Relative amounts of phospho-p38 MAPK (Ctrl: 100.0 ± 9.1%; IQ: 105.1 ± 9.5%; n = 12; p = 0.31; ΔN: 107.9 ± 8.7%; n = 12; p = 0.11) in hippocampal CA1 regions expressing BRAG1-IQ and BRAG1-ΔN. Relative amounts of total p38 MAPK (Ctrl: 100.0 ± 9.9%; IQ: 103.0 ± 10.8%; n = 12; p = 0.64; ΔN: 106.0 ± 10.7%; n = 12; p = 0.94) in hippocampal CA1 regions expressing BRAG1-IQ and BRAG1-ΔN. The relative values and SEs were normalized to average amounts of AMPA and NMDA responses from control CA1 cells, phospho-JNK, phospho-p38 MAPK, total JNK, or total p38 MAPK from control hippocampal CA1 regions. *p < 0.05 (Wilcoxon test).