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Role of PI 3-kinase, Akt and Bcl-2–related proteins in sustaining the survival of neurotrophic factor–independent adult sympathetic neurons
By adulthood, sympathetic neurons have lost dependence on NGF and NT-3 and are able to survive in culture without added neurotrophic factors. To understand the molecular mechanisms that sustain adult neurons, we established low density, glial cell-free cultures of 12-wk rat superior cervical ganglio...
Autores principales: | , , , , , |
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Formato: | Texto |
Lenguaje: | English |
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The Rockefeller University Press
2001
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2196191/ https://www.ncbi.nlm.nih.gov/pubmed/11524433 http://dx.doi.org/10.1083/jcb.200101068 |
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author | Orike, Nina Middleton, Gayle Borthwick, Emma Buchman, Vladimir Cowen, Timothy Davies, Alun M. |
author_facet | Orike, Nina Middleton, Gayle Borthwick, Emma Buchman, Vladimir Cowen, Timothy Davies, Alun M. |
author_sort | Orike, Nina |
collection | PubMed |
description | By adulthood, sympathetic neurons have lost dependence on NGF and NT-3 and are able to survive in culture without added neurotrophic factors. To understand the molecular mechanisms that sustain adult neurons, we established low density, glial cell-free cultures of 12-wk rat superior cervical ganglion neurons and manipulated the function and/or expression of key proteins implicated in regulating cell survival. Pharmacological inhibition of PI 3-kinase with LY294002 or Wortmannin killed these neurons, as did dominant-negative Class I(A) PI 3-kinase, overexpression of Ruk(l) (a natural inhibitor of Class I(A) PI 3-kinase), and dominant-negative Akt/PKB (a downstream effector of PI 3-kinase). Phospho-Akt was detectable in adult sympathetic neurons grown without neurotrophic factors and this was lost upon PI 3-kinase inhibition. The neurons died by a caspase-dependent mechanism after inhibition of PI 3-kinase, and were also killed by antisense Bcl-x(L) and antisense Bcl-2 or by overexpression of Bcl-x(S), Bad, and Bax. These results demonstrate that PI 3-kinase/Akt signaling and the expression of antiapoptotic members of the Bcl-2 family are required to sustain the survival of adult sympathetic neurons. |
format | Text |
id | pubmed-2196191 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2001 |
publisher | The Rockefeller University Press |
record_format | MEDLINE/PubMed |
spelling | pubmed-21961912008-05-01 Role of PI 3-kinase, Akt and Bcl-2–related proteins in sustaining the survival of neurotrophic factor–independent adult sympathetic neurons Orike, Nina Middleton, Gayle Borthwick, Emma Buchman, Vladimir Cowen, Timothy Davies, Alun M. J Cell Biol Article By adulthood, sympathetic neurons have lost dependence on NGF and NT-3 and are able to survive in culture without added neurotrophic factors. To understand the molecular mechanisms that sustain adult neurons, we established low density, glial cell-free cultures of 12-wk rat superior cervical ganglion neurons and manipulated the function and/or expression of key proteins implicated in regulating cell survival. Pharmacological inhibition of PI 3-kinase with LY294002 or Wortmannin killed these neurons, as did dominant-negative Class I(A) PI 3-kinase, overexpression of Ruk(l) (a natural inhibitor of Class I(A) PI 3-kinase), and dominant-negative Akt/PKB (a downstream effector of PI 3-kinase). Phospho-Akt was detectable in adult sympathetic neurons grown without neurotrophic factors and this was lost upon PI 3-kinase inhibition. The neurons died by a caspase-dependent mechanism after inhibition of PI 3-kinase, and were also killed by antisense Bcl-x(L) and antisense Bcl-2 or by overexpression of Bcl-x(S), Bad, and Bax. These results demonstrate that PI 3-kinase/Akt signaling and the expression of antiapoptotic members of the Bcl-2 family are required to sustain the survival of adult sympathetic neurons. The Rockefeller University Press 2001-09-03 /pmc/articles/PMC2196191/ /pubmed/11524433 http://dx.doi.org/10.1083/jcb.200101068 Text en Copyright © 2001, The Rockefeller University Press This article is distributed under the terms of an Attribution–Noncommercial–Share Alike–No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution–Noncommercial–Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/). |
spellingShingle | Article Orike, Nina Middleton, Gayle Borthwick, Emma Buchman, Vladimir Cowen, Timothy Davies, Alun M. Role of PI 3-kinase, Akt and Bcl-2–related proteins in sustaining the survival of neurotrophic factor–independent adult sympathetic neurons |
title | Role of PI 3-kinase, Akt and Bcl-2–related proteins in sustaining the survival of neurotrophic factor–independent adult sympathetic neurons |
title_full | Role of PI 3-kinase, Akt and Bcl-2–related proteins in sustaining the survival of neurotrophic factor–independent adult sympathetic neurons |
title_fullStr | Role of PI 3-kinase, Akt and Bcl-2–related proteins in sustaining the survival of neurotrophic factor–independent adult sympathetic neurons |
title_full_unstemmed | Role of PI 3-kinase, Akt and Bcl-2–related proteins in sustaining the survival of neurotrophic factor–independent adult sympathetic neurons |
title_short | Role of PI 3-kinase, Akt and Bcl-2–related proteins in sustaining the survival of neurotrophic factor–independent adult sympathetic neurons |
title_sort | role of pi 3-kinase, akt and bcl-2–related proteins in sustaining the survival of neurotrophic factor–independent adult sympathetic neurons |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2196191/ https://www.ncbi.nlm.nih.gov/pubmed/11524433 http://dx.doi.org/10.1083/jcb.200101068 |
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