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Detecting PTEN and PI3K Signaling in Brain

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Book cover PTEN

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1388))

Abstract

The central nervous system is comprised of multiple cell types including neurons, glia, and other supporting cells that may differ dramatically in levels of signaling pathway activation. Immunohistochemistry in conjunction with drug interference are powerful tools that allow evaluation of signaling pathways in different cell types of the mouse central nervous system in vivo. Here we provide detailed protocols for immunohistochemistry to evaluate three essential components in the PI3K pathway in mouse brain: Pten, p-Akt, and p-4ebp1, and for rapamycin treatment to modulate mTOR signaling in vivo.

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References

  1. Chalhoub N, Baker SJ (2009) PTEN and the PI3-kinase pathway in cancer. Annu Rev Pathol 4:127–150

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  2. Alessi DR et al (1997) 3-Phosphoinositide-dependent protein kinase-1 (PDK1): structural and functional homology with the Drosophila DSTPK61 kinase. Curr Biol 7(10):776–789

    Article  CAS  PubMed  Google Scholar 

  3. Sarbassov DD et al (2005) Phosphorylation and regulation of Akt/PKB by the rictor-mTOR complex. Science 307(5712):1098–1101

    Article  CAS  PubMed  Google Scholar 

  4. Sengupta S, Peterson TR, Sabatini DM (2010) Regulation of the mTOR complex 1 pathway by nutrients, growth factors, and stress. Mol Cell 40(2):310–322

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  5. Shah OJ, Hunter T (2006) Turnover of the active fraction of IRS1 involves raptor-mTOR- and S6K1-dependent serine phosphorylation in cell culture models of tuberous sclerosis. Mol Cell Biol 26(17):6425–6434

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  6. Harrington LS et al (2004) The TSC1-2 tumor suppressor controls insulin-PI3K signaling via regulation of IRS proteins. J Cell Biol 166(2):213–223

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  7. Zhu G et al (2012) Pten deletion causes mTorc1-dependent ectopic neuroblast differentiation without causing uniform migration defects. Development 139(18):3422–3431

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  8. Chalhoub N et al (2009) Cell type specificity of PI3K signaling in Pdk1- and Pten-deficient brains. Genes Dev 23(14):1619–1624

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  9. Endersby R, Baker SJ (2008) PTEN signaling in brain: neuropathology and tumorigenesis. Oncogene 27(41):5416–5430

    Article  CAS  PubMed  Google Scholar 

  10. Fraser MM et al (2004) Pten loss causes hypertrophy and increased proliferation of astrocytes in vivo. Cancer Res 64(21):7773–7779

    Article  CAS  PubMed  Google Scholar 

  11. Kwon CH et al (2001) Pten regulates neuronal soma size: a mouse model of Lhermitte-Duclos disease. Nat Genet 29(4):404–411

    Article  CAS  PubMed  Google Scholar 

  12. Groszer M et al (2001) Negative regulation of neural stem/progenitor cell proliferation by the Pten tumor suppressor gene in vivo. Science 294(5549):2186–2189

    Article  CAS  PubMed  Google Scholar 

  13. Backman SA et al (2001) Deletion of Pten in mouse brain causes seizures, ataxia and defects in soma size resembling Lhermitte-Duclos disease. Nat Genet 29(4):396–403

    Article  CAS  PubMed  Google Scholar 

  14. Sperow M et al (2012) Phosphatase and tensin homologue (PTEN) regulates synaptic plasticity independently of its effect on neuronal morphology and migration. J Physiol 590(Pt 4):777–792

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  15. Huang S, Bjornsti MA, Houghton PJ (2003) Rapamycins: mechanism of action and cellular resistance. Cancer Biol Ther 2(3):222–232

    Article  CAS  PubMed  Google Scholar 

  16. Sarbassov DD et al (2006) Prolonged rapamycin treatment inhibits mTORC2 assembly and Akt/PKB. Mol Cell 22(2):159–168

    Article  CAS  PubMed  Google Scholar 

  17. O’Reilly KE et al (2006) mTOR inhibition induces upstream receptor tyrosine kinase signaling and activates Akt. Cancer Res 66(3):1500–1508

    Article  PubMed  PubMed Central  Google Scholar 

  18. Kwon CH et al (2003) mTor is required for hypertrophy of Pten-deficient neuronal soma in vivo. Proc Natl Acad Sci U S A 100(22):12923–12928

    Article  CAS  PubMed  PubMed Central  Google Scholar 

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Acknowledgments

We thank Sherri Rankin for the p-4ebp1 IHC image. SJB was supported in part by grants from the NIH (CA096832 and CA188516) and by the American Lebanese and Syrian Associated Charities of St Jude.

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Authors and Affiliations

  1. Department of Pathology, University of Tennessee Health Sciences Center, Memphis, TN, 38163, USA

    Guo Zhu

  2. Department of Developmental Neurobiology, St Jude Children’s Research Hospital, Room D2006C, 262 Danny Thomas Place, Memphis, TN, 38105-3678, USA

    Suzanne J. Baker

Authors
  1. Guo Zhu
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  2. Suzanne J. Baker
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Corresponding author

Correspondence to Suzanne J. Baker .

Editor information

Editors and Affiliations

  1. University of Toronto, Princess Margaret Cancer Centre, Toronto, Ontario, Canada

    Leonardo Salmena

  2. University of Toronto, Princess Margaret Cancer Centre, Toronto, Ontario, Canada

    Vuk Stambolic

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© 2016 Springer Science+Business Media New York

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Zhu, G., Baker, S.J. (2016). Detecting PTEN and PI3K Signaling in Brain. In: Salmena, L., Stambolic, V. (eds) PTEN. Methods in Molecular Biology, vol 1388. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-3299-3_5

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  • DOI: https://doi.org/10.1007/978-1-4939-3299-3_5

  • Published:

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-3297-9

  • Online ISBN: 978-1-4939-3299-3

  • eBook Packages: Springer Protocols

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