(related to Figure 1). Depletion of Naa10 from CA1 does not affect global brain morphology, spatial memory, exploration or induce depression.
(A) The mRNAs of the NatA complex are enriched in the mouse hippocampus. Shown are RNA in situ hybridization images of Naa10, Naa15 and Hypk genes in the brain (sagittal section) of an 8-week-old male B6 mouse from the Allen Brain Atlas (https://mouse.brain-map.org). Colors ranging from blue to red indicate the relative expression levels from low to high, respectively. OLF, olfactory bulb; CTX, cerebral cortex; HIP, hippocampus; CB, cerebellum; BS, brain stem. The numbers of experiments conducted by the Allen Brain Atlas database are 1 for Naa10, 1 for Naa15 and 2 for HYPK (using two different antisense RNAs), respectively.
(B) The Naa10 Ab used in the study fails to detect Naa10 in the hippocampus of an 8-week-old Naa10-KO mouse. The scale bar is indicated.
(C) Naa10 protein is enriched in the hippocampal CA1 and DG of a 16-week-old mouse. The scale bar is indicated. The images shown are representative of three independent experiments.
(D) Naa10 is enriched in the mature neurons of an 8-week-old mouse. Immunohistochemistry fluorescent staining of NeuN and Nestin serves as a mature neuronal marker and a marker for neural stem cells, respectively. The scale bar is indicated.
(E) Schematic illustration of the experimental strategy for hippocampal CA1-specific Naa10 depletion. Naa10-cKO mice were generated by crossing male mice containing Camk2a-promoter-driven Cre and female mice carrying loxP-flanked Naa10 allele.
(F) Naa10 is completely depleted in CA1 in a 12-week-old Naa10-cKO mouse. The hippocampi of a control (Ctrl) and a Naa10-cKO mouse were subjected to immunohistochemistry fluorescent staining using Naa10 Ab. DAPI was used to stain DNA. The scale bar is indicated.
(G) Sagittal views of brain morphology of 2 control (Ctrl) and 3 Naa10-cKO mice by computed tomography. The scale bar is indicated.
(H) Coronal views of brain morphology in control (Ctrl) and Naa10-cKO mice by computed tomography. Representative images from 2 control and 3 Naa10-cKO mice are shown.
(I) Depletion of Naa10 from CA1 does not affect spatial memory in the Y maze test.
(J) Depletion of Naa10 from CA1 does not induce depressive-like behaviors in the tail suspension test.
(K) Depletion of Naa10 from CA1 does not result in difference in open field test.
(I-K) Shown are individual data points and mean ± SD for each group (Ctrl: 6 mice, cKO: 12 mice). ns, not significant by unpaired two-tailed t test.