QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

HOME  |   SEARCH  |   ARCHIVE  |   SUBSCRIBE  |   CONTACT  |   HELP

The Journal of Neuroscience, October 20, 2004, 24(42):9361-9371; doi:10.1523/JNEUROSCI.2365-04.2004

This Article Full Text Full Text (PDF) Submit an eLetter Alert me when this article is cited Alert me when eLetters are posted Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in Web of Science Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Web of Science (23) Citing Articles via Google Scholar Google Scholar Articles by Díaz-Hernández, M. Articles by Lucas, J. J. Search for Related Content PubMed PubMed Citation Articles by Díaz-Hernández, M. Articles by Lucas, J. J.

 Previous Article  |  Next Article 

Neurobiology of Disease
Biochemical, Ultrastructural, and Reversibility Studies on Huntingtin Filaments Isolated from Mouse and Human Brain

Miguel Díaz-Hernández,¹ Fernando Moreno-Herrero,² Pilar Gómez-Ramos,³ María A. Morán,³ Isidro Ferrer,⁴ Arturo M. Baró,² Jesús Avila,¹ Félix Hernández,¹ and José J. Lucas¹

¹Centro de Biología Molecular "Severo Ochoa", Consejo Superior de Investigaciones Científicas, ²Laboratorio de Nuevas Microscopías, Departamento de Física de la Materia Condensada, and ³Departamento de Morfología, Facultad de Medicina, Universidad Autónoma de Madrid, 28029 Madrid, Spain, and ⁴Institut de Neuropatologia, Servei d'Anatomia Patologica, Hospital Princeps d'Espanya, Hospitalet de Llobregat, 08907 Barcelona, Spain

Huntington's disease (HD) and eight additional inherited neurological disorders are caused by CAG triplet-repeat expansions leading to expanded polyglutamine-sequences in their respective proteins. These triplet-CAG repeat disorders have in common the formation of aberrant intraneuronal proteinaceous inclusions containing the expanded polyglutamine sequences. These aggregates have been postulated to contribute to pathogenesis caused by conformational toxicity, sequestration of other polyglutamine-containing proteins, or by interfering with certain enzymatic activities. Testing these hypotheses has been hampered by the difficulty to isolate these aggregates from brain. Here we report that polyglutamine aggregates can be isolated from the brain of the Tet/HD94 conditional mouse model of HD, by following a method based on high salt buffer homogenization, nonionic detergent extraction, and gradient fractionation. We then verified that the method can be successfully applied to postmortem HD brains. Immunoelectron microscopy, both in human and mouse samples, revealed that the stable component of the inclusions are mutant huntingtin-containing and ubiquitin-containing fibrils. Atomic-force microscopy revealed that these fibrils have a "beads on a string" morphology. Thus, they resemble the in vitro assembled filaments made of recombinant mutant-huntingtin, as well as the A and -synuclein amyloid protofibrils. Finally, by shutting down transgene expression in the Tet/HD94 conditional mouse model of HD, we were able to demonstrate that these filaments, although stable in vitro, are susceptible to revert in vivo, thus demonstrating that the previously reported reversal of ubiquitin-immunoreactive inclusions does not simply reflect disassembling of the inclusions into their constituent fibrils and suggesting that any associated conformational or protein-sequestration toxicity is also likely to revert.

Key words: Huntington's disease; aggregate purification; immunoelectron microscopy; atomic force microscopy; reversal; conditional mouse model

---

Received June 16, 2004; revised August 26, 2004; accepted September 3, 2004.

This article has been cited by other articles:

				J. Legleiter, G. P. Lotz, J. Miller, J. Ko, C. Ng, G. L. Williams, S. Finkbeiner, P. H. Patterson, and P. J. Muchowski Monoclonal Antibodies Recognize Distinct Conformational Epitopes Formed by Polyglutamine in a Mutant Huntingtin Fragment J. Biol. Chem., August 7, 2009; 284(32): 21647 - 21658. [Abstract] [Full Text] [PDF]

				Y. Furukawa, K. Kaneko, G. Matsumoto, M. Kurosawa, and N. Nukina Cross-Seeding Fibrillation of Q/N-Rich Proteins Offers New Pathomechanism of Polyglutamine Diseases J. Neurosci., April 22, 2009; 29(16): 5153 - 5162. [Abstract] [Full Text] [PDF]

				S. L. A. Wong, W. M. Chan, and H. Y. E. Chan Sodium dodecyl sulfate-insoluble oligomers are involved in polyglutamine degeneration FASEB J, September 1, 2008; 22(9): 3348 - 3357. [Abstract] [Full Text] [PDF]

				M. Coufal, M. M. Maxwell, D. E. Russel, A. M. Amore, S. M. Altmann, Z. R. Hollingsworth, A. B. Young, D. E. Housman, and A. G. Kazantsev Discovery of a Novel Small-Molecule Targeting Selective Clearance of Mutant Huntingtin Fragments J Biomol Screen, April 1, 2007; 12(3): 351 - 360. [Abstract] [PDF]

				M. Li, E. S. Chevalier-Larsen, D. E. Merry, and M. I. Diamond Soluble Androgen Receptor Oligomers Underlie Pathology in a Mouse Model of Spinobulbar Muscular Atrophy J. Biol. Chem., February 2, 2007; 282(5): 3157 - 3164. [Abstract] [Full Text] [PDF]

				M. L. Duennwald, S. Jagadish, P. J. Muchowski, and S. Lindquist Flanking sequences profoundly alter polyglutamine toxicity in yeast PNAS, July 18, 2006; 103(29): 11045 - 11050. [Abstract] [Full Text] [PDF]

				A. M. Ellisdon, B. Thomas, and S. P. Bottomley The Two-stage Pathway of Ataxin-3 Fibrillogenesis Involves a Polyglutamine-independent Step J. Biol. Chem., June 23, 2006; 281(25): 16888 - 16896. [Abstract] [Full Text] [PDF]

				A. G. Valera, M. Diaz-Hernandez, F. HernANdez, Z. Ortega, and J. J. Lucas The Ubiquitin-Proteasome System in Huntington's Disease Neuroscientist, December 1, 2005; 11(6): 583 - 594. [Abstract] [PDF]

				M. Diaz-Hernandez, J. Torres-Peraza, A. Salvatori-Abarca, M. A. Moran, P. Gomez-Ramos, J. Alberch, and J. J. Lucas Full Motor Recovery Despite Striatal Neuron Loss and Formation of Irreversible Amyloid-Like Inclusions in a Conditional Mouse Model of Huntington's Disease J. Neurosci., October 19, 2005; 25(42): 9773 - 9781. [Abstract] [Full Text] [PDF]

				WCHD abstracts: platform presentations J. Neurol. Neurosurg. Psychiatry, October 1, 2005; 76(suppl_4): A16 - A26. [Full Text] [PDF]

Home  |   Search  |   Archive  |   Subscribe  |   Contact  |   Help