AD transgenic mouse models had increased locomotor activity in the open field.

<p>Both APP+PS1 and Tg4510 mice displayed greater total distance travelled in the open field, compared to nontransgenic control mice, regardless of the diet. Locomotor activity was assessed during a single 15min trial in the open field. Data are presented as mean ± SEM. **p<0.005 and ***p<0.0001.</p

KD increased ketosis and reduced glucose levels.

<p>(A, C) KD (black bars) successfully increased peripheral β-hydroxybutyrate (BHB) levels after 4 weeks or 4 months, compared to a control diet (NIH-31, open bars). (B) Accordingly, circulating glucose levels were found to be decreased in KD-fed mice, in all genotypes. Glucose and BHB levels were measured utilizing a commercially available glucose/ketone monitoring system (Nova Max^© Plus). Data are presented as mean ± SEM (n=10). **p<0.0004, ***p<0.0001.</p

Astrocytosis and microglial activation were observed in Tg4510 mice.

<p>Micrograph representation (5X) of hippocampi stained for GFAP positive astrocytes (panels A-G) or Iba-1 positive microglia (panels H-N) in Ntg (A, D, H, K), APP+PS1 (B, E, I, L) and Tg4510 (C, F, J, M) mice kept on either NIH-31 (A, B, C, H, I, J) or KD diets (D, E, F, K, L, M). (G, N) Immunostaining was digitally quantified by Mirax image analysis. Data are presented as mean ± SEM. Scale bar = 500μm; inset scale bar = 100 μm ***p<0.0001.</p

Experimental design for the study of a ketogenic diet using two mouse models of Alzheimer’s pathology.

<p>APP+PS1, Tg4510 and nontransgenic littermates received either a control (NIH-31) or a low carbohydrate, medium-chain triglyceride rich, ketogenic diet (KD) for 16 weeks. Blood was collected at 4 weeks and 16 weeks for measurement of circulating ketone and glucose levels. Near the end of the administration period, mice received a variety of behavioral tests. Tissue was collected after a 16 week diet administration.</p

Changes in body weight and food intake throughout experiment.

<p>(A) Assessments of body weight and (B) food intake in APP+PS1, Tg4510 and nontransgenic mice on either control diet (NIH-31, open symbols) or ketogenic diet (KD, solid symbols) for 4 months. Both transgenic mouse lines weighed significantly less than nontransgenic mice. (B) Smaller body weights did not result from reductions in food intake. The Tg4510 mice ate significantly more food than did nontransgenic mice. Data are presented as mean ± SEM. *p<0.02, ***p<0.0001.</p

KD enhanced motor performance in all genotypes.

<p>Motor performance was assessed using two variations of the rotarod test. (A) Both genotype and diet effects were found in the standard accelerating rotarod test and (B) Average latency to fall in the accelerating rotarod was significantly greater in mice fed a KD. (C) Similarly, the ketogenic diet significantly enhanced motor performance in the non-accelerating variation of the test in all genotypes. Overall, latency to fall was greater in nontransgenic mice than in either APP+PS1 or Tg4510 mice. Data are presented as mean ± SEM. **p<0.007.</p

KD did not rescue neuronal loss in Tg4510 mice.

<p>Micrograph representation (2X) of neuronal staining (NeuN) in Ntg (A, D), APP+PS1 (B, E) and Tg4510 (C, F) mice kept on either NIH-31 (A, B, C) or KD diets (D, E, F). (G) Immunoreactivity for the neuronal marker NeuN was significantly reduced in Tg4510 mice compared to both nontransgenic and APP+PS1 mice. (H) Hippocampal volume (expressed in mm³) was calculated in Nissl stained sections. In agreement with the neuronal loss observed, hippocampal volume was significantly smaller in Tg4510 line, compared to the other genotypes. No diet effects were observed. Immunostaining was digitally quantified by Mirax image analysis. Data are presented as mean ± SEM. 2X Scale bar = 1000μm; 20X inset scale bar = 100 μm. **p<0.01 and ***p<0.0001.</p

KD did not rescue spatial memory deficits.

<p>Spatial memory was assessed by the 2-day radial arm water maze (RAWM). (A) Both APP+PS1 and Tg4510 (tau) mice made more errors on the 2-day RAWM compared to Ntg control mice, regardless of the diet. (B) APP+PS1 and Tg4510 mice consistently made more entries into the wrong arms, suggesting that neither were able to learn a new platform location on the reversal trial. Data are presented as mean ± SEM. **p<0.01 and ***p<0.0001.</p

Study 3: CD68 (A-C) and CD45 (D-F) immunostaining is observed throughout the hippocampus of study mice.

<p>In the hippocampus, no significant differences are observed in the amount of total CD68 staining between the NEP-n, NEP-s, and NEP-m groups, but CD45 staining in NEP-s treated mice is greater than CD45 staining in NEP-m treated mice. Scale bar = 50 µm. Panels G and H present ANOVA analysis of the ratio of CD68 to congophilic staining, and of the ratio of quantitated CD45 to congophilic staining, respectively, in the hippocampus of study mice. The (*) indicates significance compared to NEP-m mice with p<0.05, and the (^∧) indicates significance compared to Tg control mice with p<0.05 or p<0.01 (°).Tg control (n = 9), NEP-n (n = 4), NEP-m (n = 7), NEP-s (n = 6).</p

Study 1: Distribution of HA expression in the hippocampus (A–D) and frontal cortex (E–H) following intracranial administration of rAAV vectors, detected using an anti-HA antibody.

<p>Panels A & E show NEP-n treated animals; panels B & F show NEP-m treated animals; panels C & G show NEP-s treated animals. Panels D & H show no positive staining in the contralateral uninjected left hippocampus and left anterior cortex, respectively. Scale bar = 120 µm.</p