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Does Notch Act Through mTOR to Regulate Protein Synthesis?
Author(s) -
Huot Joshua,
Thompson Brian,
McMullen Charlotte,
Arthur Susan
Publication year - 2018
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.2018.32.1_supplement.768.9
Subject(s) - notch signaling pathway , pi3k/akt/mtor pathway , anabolism , protein biosynthesis , chemistry , protein synthesis inhibitor , fusion protein , microbiology and biotechnology , signal transduction , medicine , biology , biochemistry , cycloheximide , recombinant dna , gene
PURPOSE Notch signaling is thought to be crucial in regulating skeletal muscle regeneration, however, the impact Notch signaling has on other skeletal muscle processes (e.g. protein synthesis) remains unclear. The purpose of this project was to determine the effects of Notch inhibition on protein synthesis in C2C12 cells. METHODS C2C12s were seeded, proliferated to ~ 90% confluence and then differentiated for four days. At the onset of differentiation, C2C12s were treated every 12 hours (from 0hr–96hr) with one of the following conditions: 4 μmoL of gamma secretase inhibitor (GSI: Notch inhibitor), 100 nmoL of Rapamycin (mechanistic target of rapamycin (mTOR) inhibitor), both, or control. Four days post‐differentiation C2C12s were fixed for immunofluorescence to quantify myotube formation, or were collected and analyzed for components of Notch, anabolic signaling, and protein synthesis (via puromycin incorporation). RESULTS GSI treatment increased fusion index compared to the other three treatments (fusing nuclei/total nuclei) (p <0.0001). Rapamycin abrogated fusion index relative to control (p <0.0001). GSI + Rapamycin‐treated CC12s also had reduced fusion index compared to control (p<0.0001). GSI‐treated C2C12s displayed elevated protein synthesis compared to all other groups (p < 0.05). Rapamycin reduced protein synthesis compared to control (p < 0.05). GSI+Rapamycin‐treated C2C12s did not differ in protein synthesis compared to control or Rapamycin. CONCLUSIONS Our data suggests that Notch may regulate protein synthesis through mTOR. This abstract is from the Experimental Biology 2018 Meeting. There is no full text article associated with this abstract published in The FASEB Journal .