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GSK-3 signaling in developing cortical neurons is essential for radial migration and dendritic orientation.

Morgan-Smith M, Wu Y, Zhu X, Pringle J, Snider WD - Elife (2014)

Bottom Line: Radial migration in hippocampus was similarly affected.GSK-3 regulation of migration in neurons was independent of Wnt/β-catenin signaling.Importantly, phosphorylation of the migration mediator, DCX, at ser327, and phosphorylation of the semaphorin signaling mediator, CRMP-2, at Thr514 were markedly decreased.

View Article: PubMed Central - PubMed

Affiliation: UNC Neuroscience Center, University of North Carolina, Chapel Hill, United States Neurobiology Curriculum, University of North Carolina, Chapel Hill, United States meghan_morgan@med.unc.edu.

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Phosphorylation status of GSK-3 substrates.(A–A') Western blots of P0 cortical lysates from Gsk3loxp:Neurod6 mutants and wild-type controls performed in triplicate. Levels of GSK-3 proteins and phospho-target proteins are shown. Strong reductions in phosphorylation of doublecortin on ser327/Thr321 and CRMP-2 on Thr514 are evident. (A') Quantification of relative densities from A. p values shown in figure (n = 3, unpaired t-test) (B–B') Western blots of cortical lysates at P0 showing levels of other GSK-3 targets. No changes were observed in phosphorylation of dynamin, pCREB, or pFAK. No change was observed in cleaved caspase-3. GAPDH was used as a loading control. (B') Quantification of relative protein densities (n = 3, unpaired t-test).DOI:http://dx.doi.org/10.7554/eLife.02663.017
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fig7: Phosphorylation status of GSK-3 substrates.(A–A') Western blots of P0 cortical lysates from Gsk3loxp:Neurod6 mutants and wild-type controls performed in triplicate. Levels of GSK-3 proteins and phospho-target proteins are shown. Strong reductions in phosphorylation of doublecortin on ser327/Thr321 and CRMP-2 on Thr514 are evident. (A') Quantification of relative densities from A. p values shown in figure (n = 3, unpaired t-test) (B–B') Western blots of cortical lysates at P0 showing levels of other GSK-3 targets. No changes were observed in phosphorylation of dynamin, pCREB, or pFAK. No change was observed in cleaved caspase-3. GAPDH was used as a loading control. (B') Quantification of relative protein densities (n = 3, unpaired t-test).DOI:http://dx.doi.org/10.7554/eLife.02663.017

Mentions: To determine the status of relevant GSK-3 targets, we conducted western blot analysis of GSK-3 substrates that have been implicated in migration regulation. Recently, the key migration regulator doublecortin (DCX) was shown to be a GSK-3 substrate (Bilimoria et al., 2010). In P0 cortical lysates of Gsk3loxp:Neurod6 mutants, we observed a 61% decrease in phosphorylated DCX on Ser327 (Figure 7A–A'). These results demonstrate that DCX is importantly regulated by GSK-3 in developing cortical neurons.10.7554/eLife.02663.017Figure 7.Phosphorylation status of GSK-3 substrates.


GSK-3 signaling in developing cortical neurons is essential for radial migration and dendritic orientation.

Morgan-Smith M, Wu Y, Zhu X, Pringle J, Snider WD - Elife (2014)

Phosphorylation status of GSK-3 substrates.(A–A') Western blots of P0 cortical lysates from Gsk3loxp:Neurod6 mutants and wild-type controls performed in triplicate. Levels of GSK-3 proteins and phospho-target proteins are shown. Strong reductions in phosphorylation of doublecortin on ser327/Thr321 and CRMP-2 on Thr514 are evident. (A') Quantification of relative densities from A. p values shown in figure (n = 3, unpaired t-test) (B–B') Western blots of cortical lysates at P0 showing levels of other GSK-3 targets. No changes were observed in phosphorylation of dynamin, pCREB, or pFAK. No change was observed in cleaved caspase-3. GAPDH was used as a loading control. (B') Quantification of relative protein densities (n = 3, unpaired t-test).DOI:http://dx.doi.org/10.7554/eLife.02663.017
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Related In: Results  -  Collection

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fig7: Phosphorylation status of GSK-3 substrates.(A–A') Western blots of P0 cortical lysates from Gsk3loxp:Neurod6 mutants and wild-type controls performed in triplicate. Levels of GSK-3 proteins and phospho-target proteins are shown. Strong reductions in phosphorylation of doublecortin on ser327/Thr321 and CRMP-2 on Thr514 are evident. (A') Quantification of relative densities from A. p values shown in figure (n = 3, unpaired t-test) (B–B') Western blots of cortical lysates at P0 showing levels of other GSK-3 targets. No changes were observed in phosphorylation of dynamin, pCREB, or pFAK. No change was observed in cleaved caspase-3. GAPDH was used as a loading control. (B') Quantification of relative protein densities (n = 3, unpaired t-test).DOI:http://dx.doi.org/10.7554/eLife.02663.017
Mentions: To determine the status of relevant GSK-3 targets, we conducted western blot analysis of GSK-3 substrates that have been implicated in migration regulation. Recently, the key migration regulator doublecortin (DCX) was shown to be a GSK-3 substrate (Bilimoria et al., 2010). In P0 cortical lysates of Gsk3loxp:Neurod6 mutants, we observed a 61% decrease in phosphorylated DCX on Ser327 (Figure 7A–A'). These results demonstrate that DCX is importantly regulated by GSK-3 in developing cortical neurons.10.7554/eLife.02663.017Figure 7.Phosphorylation status of GSK-3 substrates.

Bottom Line: Radial migration in hippocampus was similarly affected.GSK-3 regulation of migration in neurons was independent of Wnt/β-catenin signaling.Importantly, phosphorylation of the migration mediator, DCX, at ser327, and phosphorylation of the semaphorin signaling mediator, CRMP-2, at Thr514 were markedly decreased.

View Article: PubMed Central - PubMed

Affiliation: UNC Neuroscience Center, University of North Carolina, Chapel Hill, United States Neurobiology Curriculum, University of North Carolina, Chapel Hill, United States meghan_morgan@med.unc.edu.

Show MeSH