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. 2018 Nov 5;217(11):4007–4024. doi: 10.1083/jcb.201803105

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© 2018 Liu et al.

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Figure 8. — EphB kinase activation promotes MF terminal maturation. (A) Transmission electron micrographs from PW3 mice show MF terminals (red) and postsynaptic spines (green) in synapses with giant bouton. Asterisks mark PSD. Bars, 0.5 µm. (B) The decreased PSD number per spine and docked vesicles of MF synapses from SPB (281–312 synapses) or IPB (37–46 synapses) regions in Lnx1^−/− mice were restored to normal level in Lnx1^−/−; EphB2 ^F620D/F620D mice. n = 3–4 mice per group. (C) Transmission electron micrographs from PW3 mice show axon terminals (red) and postsynaptic spines (green) in typical synapses with regular bouton. Asterisks mark PSD. Bars, 0.5 µm. (D) The number of vesicles per terminal in typical synapses was significantly correlated with the PSD length, determined by EM analysis in PW3 WT (r² = 0.62; P < 0.001), EphB2 ^F620D/F620D (r² = 0.42; P < 0.001), and Lnx1^−/−; EphB2 ^F620D/F620D (r² = 0.40; P < 0.001) mice but not in Lnx1^−/− mice (left). The area of vesicles per terminal in typical synapses was significantly correlated with PSD length in PW3 WT (r² = 0.66; P < 0.001), EphB2 ^F620D/F620D (r² = 0.54; P < 0.001), and Lnx1^−/−; EphB2 ^F620D/F620D (r² = 0.59; P < 0.001) mice but not in Lnx1^−/− mice (right). A dot presents a synapse. (E) EM analysis showed that the decreased vesicle density in terminals that formed synapses in Lnx1^−/− mice was restored to normal level in Lnx1^−/−; EphB2 ^F620D/F620D mice. (F) The increased average distance of vesicles to the presynaptic membrane in Lnx1^−/− mice was restored to normal level in Lnx1^−/−; EphB2 ^F620D/F620D mice. n = 72–165 synapses per group. n = 3–4 mice per group in D–F. (G) Representative average traces (left) and summary graph (right) showed that increased PPRs upon SPB stimulation in PW3 Lnx1^−/− mice were restored to normal level in Lnx1^−/−; EphB2 ^F620D/F620D mice. Bar, 50 ms. n = 22–27 neurons from three to four mice per group. (H) Proposed model for postsynaptic Lnx1–EphB complex–mediated retrograde regulation of MF axon terminal maturation. Lnx1 binds and stabilizes postsynaptic EphB receptors in hippocampal CA3 pyramidal neurons to sculpt postsynaptic structure, which helps to guide MF axon targeting and promote terminal maturation through a trans-synaptic regulation. Means ± SEM; one-way ANOVA with Tukey’s post hoc test (B and E–G); *, P < 0.05; **, P < 0.01; ***, P < 0.001.