Fig. 3.
Neuronal rescue of NCLX expression impedes cognitive decline and AD pathology. a Schematic of tetracycline-responsive (TRE) transgenic construct and neuronal-specific driver, Camk2a-tTA. Neuronal-specific gain-of-function models were crossed with 3xTg-AD mutant mice to generate 3xTg-AD × TRE-NCLX × Camk2a-tTA mice. b NCLX mRNA expression corrected to the housekeeping gene, Rps13, expressed as fold change vs. tTA controls, n = 3 for all groups. c Western blots for NCLX expression and mitochondrial calcium uniporter channel components, tissue isolated from the hippocampus of 2 months old mice. d, e Y-maze spontaneous alternation test. d Percentage spontaneous alternation, e Total number of arm entries. f–h Fear-conditioning test. f Freezing responses in the training phase, g contextual recall freezing responses, h Cued recall freezing responses. i, j Soluble and insoluble Aβ1–40 and Aβ1–42 levels in brain cortex of 12 months old mice. k Representative immunohistochemical staining for 4G8-reactive β-amyloid; 4 × scale bar = 100 μm, 40 × scale bar = 50 μm. l Quantification of the integrated optical density area for Aβ immunoreactivity, n = 4 for all groups. m Western blots of full-length APP, ADAM-10, BACE1, PS1, Nicastrin, APH, and tubulin (loading control) in cortex homogenate of 12 months old mice, n = 3 for all groups. n Representative western blots of soluble and insoluble total tau (HT7), phosphorylated tau at residues S202/T205 (AT8), T231/S235 (AT180), T181 (AT270), and S396 (PHF13) in soluble brain cortex homogenate of 12 months old mice, n = 3 for all groups. o Representative hippocampal staining for total tau (HT7) and phospho-tau S202/T205 (AT8) immunoreactivity in 12 months old mice; scale bar = 50 μm. p, q Quantification of HT7 and AT8 integrated optical density area correct to 3xTg-AD × Camk2a-tTA controls, n = 4 for all groups. (n = individual dots shown for each group in all graphs. All data presented as mean ± SEM; ****p < 0.001, **p < 0.01, *p < 0.05; one-way ANOVA with Sidak's multiple comparisons test.) Source data are available as a Source Data file