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Polyglutamine expansions cause decreased CRE-mediated transcription and early gene expression changes prior to cell death in an inducible cell model of Huntington's disease

Polyglutamine expansions cause decreased CRE-mediated transcription and early gene expression changes prior to cell death in an inducible cell model of Huntington's disease
Polyglutamine expansions cause decreased CRE-mediated transcription and early gene expression changes prior to cell death in an inducible cell model of Huntington's disease
Huntington’s disease (HD) is one of 10 known diseases caused by a (CAG)n trinucleotide repeat expansion that is translated into an abnormally long polyglutamine tract. We have developed stable inducible neuronal (PC12) cell lines that express huntingtin exon 1 with varying CAG repeat lengths under doxycycline (dox) control. The expression of expanded repeats is associated with aggregate formation, caspase-dependent cell death and decreased neurite outgrowth. Post-mitotic cells expressing mutant alleles were more prone to cell death compared with identical cycling cells. To determine early metabolic changes induced by this mutation in cell models, we studied changes in gene expression after 18 h dox induction, using Affymetrix arrays, cDNA filters and adapter-tagged competitive PCR (ATAC-PCR). At this time point there were low rates of inclusion formation, no evidence of mitochondrial compromise and no excess cell death in the lines expressing expanded compared with wild-type repeats. The expression profiles suggest novel targets for the HD mutation and were compatible with impaired cAMP response element (CRE)-mediated transcription, which we confirmed using CRE-luciferase reporter assays. Reduced CRE-mediated transcription may contribute to the loss of neurite outgrowth and cell death in polyglutamine diseases, as these phenotypes were partially rescued by treating cells with cAMP or forskolin.
1829-1845
Wyttenbach, A.
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Swartz, J.
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Kita, H.
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Thykjaer, T.
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Carmichael, J.
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Bradley, J.
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Brown, R.
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Maxwell, M.
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Schapira, A.
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Orntoft, T.F.
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Kata, K.
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Rubinsztein, D.C.
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Wyttenbach, A.
69846a0f-fb60-4a28-84eb-ed865a5e31fa
Swartz, J.
c6b73317-135f-4e75-8166-8dcb2ae4a92b
Kita, H.
73ecaabf-90c3-4563-a58a-f882a2935309
Thykjaer, T.
688f67bb-479f-4bd7-b8fc-5224bd1937dd
Carmichael, J.
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Bradley, J.
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Brown, R.
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Maxwell, M.
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Schapira, A.
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Orntoft, T.F.
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Kata, K.
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Rubinsztein, D.C.
7bc49386-aa67-4658-a966-0d952e6fd731

Wyttenbach, A., Swartz, J., Kita, H., Thykjaer, T., Carmichael, J., Bradley, J., Brown, R., Maxwell, M., Schapira, A., Orntoft, T.F., Kata, K. and Rubinsztein, D.C. (2001) Polyglutamine expansions cause decreased CRE-mediated transcription and early gene expression changes prior to cell death in an inducible cell model of Huntington's disease. Human Molecular Genetics, 10 (17), 1829-1845. (doi:10.1093/hmg/10.17.1829).

Record type: Article

Abstract

Huntington’s disease (HD) is one of 10 known diseases caused by a (CAG)n trinucleotide repeat expansion that is translated into an abnormally long polyglutamine tract. We have developed stable inducible neuronal (PC12) cell lines that express huntingtin exon 1 with varying CAG repeat lengths under doxycycline (dox) control. The expression of expanded repeats is associated with aggregate formation, caspase-dependent cell death and decreased neurite outgrowth. Post-mitotic cells expressing mutant alleles were more prone to cell death compared with identical cycling cells. To determine early metabolic changes induced by this mutation in cell models, we studied changes in gene expression after 18 h dox induction, using Affymetrix arrays, cDNA filters and adapter-tagged competitive PCR (ATAC-PCR). At this time point there were low rates of inclusion formation, no evidence of mitochondrial compromise and no excess cell death in the lines expressing expanded compared with wild-type repeats. The expression profiles suggest novel targets for the HD mutation and were compatible with impaired cAMP response element (CRE)-mediated transcription, which we confirmed using CRE-luciferase reporter assays. Reduced CRE-mediated transcription may contribute to the loss of neurite outgrowth and cell death in polyglutamine diseases, as these phenotypes were partially rescued by treating cells with cAMP or forskolin.

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Published date: 1 August 2001

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Local EPrints ID: 35633
URI: http://eprints.soton.ac.uk/id/eprint/35633
PURE UUID: 352f46b2-ffdd-46ba-9b1b-e14d04141b1a

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Date deposited: 22 May 2006
Last modified: 15 Mar 2024 07:53

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Contributors

Author: A. Wyttenbach
Author: J. Swartz
Author: H. Kita
Author: T. Thykjaer
Author: J. Carmichael
Author: J. Bradley
Author: R. Brown
Author: M. Maxwell
Author: A. Schapira
Author: T.F. Orntoft
Author: K. Kata
Author: D.C. Rubinsztein

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