Figure - PMC

Skip to main content

An official website of the United States government

Here's how you know

Here's how you know

Official websites use .gov
A .gov website belongs to an official government organization in the United States.

Secure .gov websites use HTTPS
A lock ( ) or https:// means you've safely connected to the .gov website. Share sensitive information only on official, secure websites.

View full-text article in PMC

. 2014 Apr 23;7:30. doi: 10.3389/fnmol.2014.00030

Search in PMC
Search in PubMed
View in NLM Catalog
Add to search

Copyright © 2014 Fricano, DeSpenza, Frazel, Li, O'Malley, Westbrook and Luikart.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

PMC Copyright notice

Rapamycin Inhibits Neuronal Hypertrophy and Increased p-S6 signaling Caused by Pten Knockdown (A) Pten knockdown neurons treated with fatty acids (green) have an increase in soma size and p-S6 intensity (blue) compared to neighboring control neurons (red). (B) Treating these fatty acid exposed, Pten knockdown neurons with rapamycin (green) inhibits this increase in soma size and increase in p-S6 staining (blue) compared to control neurons (red). (C) In vehicle treated neurons, there is still an increase in soma size when comparing the knockdown neurons to control neurons (Vehicle control = 94.13 ± 1.36, Vehicle shPten = 107.4 ± 1.52, p < 0.001, n = 134 and 188 neurons from 4 animals). There is also still an increase in soma size when comparing the knockdown neurons to the control neurons in the rapamycin treated cells (Veh control + rapa = 91.03 ± 0.86, Veh shPten + rapa = 99.36 ± 1.00, p < 0.001, n = 220 and 272 cells from 4 animals). However, there is no difference between the Veh shPten + rapa cells and the Veh control cells (p > 0.05), showing that rapamycin was able to rescue the soma size of knockdown cells to that of the untreated controls. There is also a significant difference between Veh shPten cells and the Veh shPten + rapa cells (p < 0.001, right panel), demonstrating that rapamycin treatment reduced the size of Pten knockdown neurons. (D) Like the vehicle, there is also still a significant increase in soma size in fatty acid treated Pten knockdown neurons compared to their controls in the rapamycin treated and non-treated groups (FA control = 96.86 ± 1.04, FA shPten = 116.6 ± 1.50, p < 0.001, n = 190 and 277 neurons from 4 animals; FA control + rapa = 87.23 ± 0.94, FA shPten + rapa = 97.38 ± 1.23, p < 0.001, n = 207 and 264 neurons from 4 animals). There was no significant difference between fatty acid treated control neurons and fatty acid treated Pten knockdown neurons treated with rapamycin, demonstrating that even with fatty acid treatment, rapamycin is still able to rescue the soma size of knockdown neurons to that of controls (FA control vs. FA shPten + rapa, p > 0.05). Finally, there was a significant difference between fatty acid treated Pten knockdown neurons and fatty acid treated Pten knockdown neurons treated with rapamycin (FA shPten vs. FA shPten + rapa, p < 0.05, right panel), demonstrating that rapamycin is able to rescue the soma size of fatty acid treated Pten knockdown neurons. (E) In vehicle treated animals, there is an increase in p-S6 signaling in knockdown neurons compared to controls (Veh control = 1.00 ± 0.02, Veh shPten = 1.28 ± 0.03, p < 0.001, n = 246 and 232 neurons from 6 animals). With rapamycin treatment, there is a reversal of this effect and a robust decrease in p-S6 signaling in the knockdown cells (Veh control + rapa = 1.00 ± 0.02, Veh shPten + rapa = 0.89 ± 0.02, p < 0.001, n = 220 and 272 neurons from 4 animals). Therefore, there is a significant difference in p-S6 signaling in vehicle rapamycin treated animals and untreated animals (p < 0.001). (F) Again, there is an increase in p-S6 signaling in fatty acid treated knockdown neurons compared to controls (fatty acid control = 1.0 ± 0.02, fatty acid shPten = 1.54 ± 0.05, n = 169 and 268 neurons from 5 animals). However, even with fatty acid treatment, there is still a significant decrease in p-S6 staining in rapamycin treated knockdown neurons compared to controls (FA control + rapa = 1.00 ± 0.01, FA shPten + rapa = 0.85 ± 0.02, p < 0.01, n = 207 and 264 neurons from 4 animals). Therefore, there is a significant decrease in p-S6 signaling in fatty acid and rapamycin treated neurons compared to fatty acid treatment alone (right, p < 0.001).