Figure 5: CaMKv knockdown in vivo reduces spine density and impairs basal synaptic neurotransmission and L-LTP.
From: The pseudokinase CaMKv is required for the activity-dependent maintenance of dendritic spines
(a) Representative immunostained coronal sections from mice subjected to bilateral lentivirus injections (scale bar, 500 μm). (b,c) The indicated GFP-positive CA1 region was dissected to examine CaMKv levels. CaMKv knockdown (shCaMKv) in the hippocampal CA1 region significantly reduced CaMKv expression (Con: n=4 mice, shCaMKv: n=6 mice, *P<0.05, Student’s t-test). (d) Hippocampal CA1 pyramidal neurons were transduced by control or lentiviral vectors expressing CaMKv shRNA. CaMKv knockdown in vivo reduced CaMKv expression (red) in CA1 neurons (scale bar, 25 μm; arrowheads: GFP-positive neurons; dotted area: magnified images of infected neurons). (e,f) CaMKv knockdown in vivo significantly reduced dendritic spine density. (scale bar, 20 μm; Con: n=34 dendrites, shCaMKv: n=21 dendrites, ***P<0.001, Student’s t-test; green; GFP, red: CaMKv, blue: 4,6-diamidino-2-phenylindole (DAPI)). (g) CaMKv knockdown in hippocampal CA1 neurons inhibited basal synaptic neurotransmission. The input–output curve was altered in the infected CA1 region after CaMKv knockdown (Con: n=8 mice, shCaMKv: n=9 mice). (h) CaMKv knockdown impaired the L-LTP. (i) The field excitatory postsynaptic potential (fEPSP) amplitude (normalized to baseline) was significantly reduced in hippocampi injected with CaMKv shRNA (Con: n=8 mice, shCaMKv: n=9 mice, *P<0.05, Student’s t-test).
