Skip to main content
. 2016 Apr 7;76(12):1308–1327. doi: 10.1002/dneu.22392

Figure 5.

Figure 5

Rapamycin treatment affects basal dendrites of postnatally born OB neurons. (A) Western blot analysis of phosphorylated ribosomal protein S6 (P‐S6, Ser235/236), total S6, phosphorylated Akt (P‐Akt, Ser473) and total Akt levels in protein lysates obtained from olfactory bulbs from animals treated with rapamycin (rapa) or vehicle solution (control). Mouse pups were treated with two or three doses of 1 mg/kg rapamycin from 8 (3 doses) or 10 days (2 doses) after birth. Doses were administered at 48 h intervals. Tubulin (Tub) was used as a loading control. (B) Quantitative analysis of cell soma volume of GFP‐positive neuronal cell body in olfactory bulb (OB). Mice were electroporated with a GFP‐encoding plasmid (pCx‐EGFP‐N1) on P1‐P2, treated with 2 doses of rapamycin or vehicle as described in A, and sacrificed at 14 dpe. All OB slices were immunostained for GFP. Neuron cell somas were three‐dimensionally reconstructed. The results are presented as a mean value ± SEM. Number of animals (N) = 5 (control), 5 (rapa). Number of analyzed cells (n) = 47 (control), 42 (rapa). ns, not significant (Mann‐Whitney test). (C) Representative three‐dimensional reconstructions of GFP‐positive neurons in olfactory bulb after IP injection of rapamycin vs. control. Scale bar = 50 µm. Mice were electroporated with pCx‐EGFP‐N1 on P1‐P2, intraperitoneally (IP) injected with two doses of rapamycin starting 10 dpe, and sacrificed at 14 dpe. (D, E) Quantification of the total number of dendritic tips (TNDT, D) and total dendritic length (TDL, E) of basal dendrites after IP rapamycin injections. (F, G) Quantification of the total number of dendritic tips (TNDT, F) and total dendritic length (TDL, G) of apical dendrites after IP rapamycin injections. The results are presented as a mean value ± SEM. For all graphs: number of individual experiments = 2; number of animals (N) = 5 (control), 5 (rapa); number of analyzed cells (n) = 52 (control), 57 (rapa). **p ≤ 0.01; ns, not significant (Mann–Whitney test). (H, I) Sholl analysis of basal and apical dendrites in OB neurons. Significance for particular radii was assessed using one‐way ANOVA. For all graphs: number of individual experiments = 2; number of animals (N) = 5 (control), 5 (rapamycin); number of analyzed cells (n): 52 (control), 57 (rapamycin). *p ≤ 0.05. [Color figure can be viewed at wileyonlinelibrary.com.]