However, in contrast to enriched wild-type mice and to nonenriched mutant mice, recovery from anisomycin-induced AZ loss was dramatically impaired: AZ densities had failed to recover at 48 hr, recovery was only about 50% at 4 days, and about Autophagy inhibitor chemical structure 90% at 8 days ( Figure 5A, yellow trace). This dramatic impairment in the reassembly of AZs upon anisomycin in enriched mice was completely rescued upon lentiviral transduction of GFP-β-Adducin into the dentate gyrus of adult β-Adducin−/− mice ( Figure 5A, red trace). Taken together, these results suggest that upon environmental enrichment, but not under control conditions, the assembly

of labile synapses is majorly compromised in the absence of β-Adducin. To determine whether impaired assembly of labile synapses in the absence of β-Adducin may affect enrichment-induced synapse gains, we analyzed LMTs and CA1 spines in β-Adducin−/− mice upon 4 weeks of enrichment. In stark contrast to wild-type mice, 4 weeks of environmental enrichment failed to increase AZ densities per LMT in β-Adducin−/− mice ( Figure 5B). This was specifically due to the local absence of β-Adducin in mossy fibers and not to a failure to respond to enriched conditions in the mutant mice, because lentiviral rescue with GFP-β-Adducin into the dentate gyrus of

β-Adducin−/− mice produced elevated AZ densities indistinguishable from those in wild-type mice in transduced neurons ( Figure 5B). A comparison of AZ CT99021 price Parvulin densities/LMT volume and LMT complexities revealed that upon environmental enrichment the more complex LMTs exhibited relatively lower contents of AZs in β-Adducin−/− mice ( Figure 5C). A detailed electron microscopic

analysis of AZ distributions, and of their relationship to postsynaptic thorny excrescences revealed that while, as reported previously ( Gogolla et al., 2009), these densities were not affected upon enrichment in wild-type mice, 4 weeks of environmental enrichment reduced AZ densities per thorn area in β-Adducin−/− mice to 58% of control values ( Figure 5D). By contrast, the increase in thorny excrescence densities upon 4 weeks of enrichment was not impaired in the β-Adducin−/− mice ( Figure 5D). More than 95% of AZs faced postsynaptic densities, and thus represented bona fide synapses. Therefore, 4 weeks of environmental enrichment produced a comparable growth of postsynaptic thorny excrescences in wild-type and β-Adducin−/− mice, but in the mutant mice this increase in thorny spine structures was not matched by a corresponding increase in the number of actual synapses. We also analyzed frequencies of satellite LMTs upon 4 weeks of enrichment. As shown in a previous study ( Gogolla et al., 2009), satellite frequencies increased substantially upon 4 weeks of enriched environment in wild-type mice ( Figure 5E).