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(American Journal of Pathology. 1998;153:695-701.)
© 1998 American Society for Investigative Pathology

Short Communications

Nonneural Nuclear Inclusions of Androgen Receptor Protein in Spinal and Bulbar Muscular Atrophy

Mei Li* , Yuji Nakagomi , Yasushi Kobayashi* , Dianne E. Merry , Fumiaki Tanaka* , Manabu Doyu* , Terunori Mitsuma§ , Yoshio Hashizume¶ , Kenneth H. Fischbeck and Gen Sobue*

From the Department of Neurology,* Nagoya University School of Medicine, Nagoya, Japan; Laboratory of Electron Microscopy, Aichi Medical University, Aichi, Japan; Department of Neurology, University of Pennsylvania School of Medicine, Philadelphia, Pennsylvania; and Department of Internal Medicine§ and Institute for Medical Science of Aging,^¶ Aichi Medical University, Aichi, Japan

	Abstract

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Spinal and bulbar muscular atrophy is an X-linked motor neuronopathy caused by the expansion of an unstable CAG repeat in the coding region of the androgen receptor (AR) gene. Nuclear inclusions of the mutant AR protein have been shown to occur in the spinal motor neurons of spinal and bulbar muscular atrophy (Li M, Kobayashi Y, Merry D, Tanaka F, Doyu M, Hashizume Y, Fischbeck KH, Sobue G: Nuclear inclusions in spinal and bulbar muscular atrophy. Ann Neurol 1998 (in press)). In this study, we demonstrate the tissue-specific distribution, immunochemical features, and fine structure of nuclear inclusions of spinal and bulbar muscular atrophy. Nuclear inclusions were observed in affected spinal and brainstem motor neurons, but not in other, nonaffected neural tissues. Similar nuclear inclusions occurred in nonneural tissues including scrotal skin, dermis, kidney, heart, and testis, but not in the spleen, liver, and muscle. These inclusions had similar epitope features detectable by antibodies that recognize a small portion of the N-terminus of the AR protein only, and they were ubiquitinated. Electron microscopic immunohistochemistry showed dense aggregates of AR-positive granular material without limiting membrane, both in the neural and nonneural inclusions. These findings indicate that nuclear inclusions of AR protein are present in selected nonneural tissues as well as in neurons that degenerate in spinal and bulbar muscular atrophy, suggesting that a common mechanism underlies in the formation of neural and nonneural nuclear inclusions.

	Introduction

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Other disorders caused by CAG repeat expansion include Huntington's disease (HD),⁹ dentatorubral-pallidoluysian atrophy (DRPLA),^10,11 Machado-Joseph disease (MJD),¹² and spinocerebellar ataxia type 1 (SCA1),¹³ type 2 (SCA2)^14–16, type 6 (SCA6),¹⁷ and type 7 (SCA7).¹⁸ These disorders share several characteristics that are likely relevant to a common pathological mechanism leading to selective neuronal loss. The mechanism is thought to be a toxic gain of function of the mutant gene products^19,20 involving cell-specific protein-protein or protein-nucleic acid interactions with the products of the mutant genes.^21-26 Intranuclear inclusions of the mutant proteins have recently been documented in the neurons of HD,^27,28 MJD,^29,30 SCA1,^26,31 DRPLA,³ and SBMA motor neurons,³³ as well as in the transgenic models of HD^34,35 and SCA1.²⁶ In all of these disorders, the inclusions can be labeled with antibodies (Abs) to the disease protein product and to ubiquitin. The inclusions are so far detected in the neurons of the affected brain areas of each disease, and rarely in other brain regions,^27-32 despite the ubiquitous expression of the disease gene product.³⁶ Furthermore, in HD, there is a correlation between increasing CAG repeat length and increasing density of the inclusions.³⁶ Thus, intranuclear inclusions of mutant protein mediated by polyglutamine-directed aggregation are thought to have a primary pathogenic role in neuronal loss for these CAG repeat diseases.^26-35

In this study, we demonstrate that nuclear inclusions of mutant AR protein occur in selected nonneural tissues as well as neural tissues in SBMA.

	Materials and Methods

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Postmortem Tissues from SBMA and Control Subjects

Various portions of brain, spinal cord, peripheral nerve, muscle, and nonneural visceral organs were sampled from five autopsied patients with SBMA (three of whom were processed for frozen samples and all five fixed in formalin). These patients were 54 to 82 years of age at death, and each showed a typical clinical phenotype of SBMA, with dysphagia, bulbar and extremity muscle weakness, and atrophy with fasciculation. Gynecomastia and diabetes mellitus were present in four patients. Duration from onset to death was 9 to 23 years, and the causes of death were empyema, bronchiectasis, and gastric cancer. The CAG repeat lengths of the AR gene determined in blood samples were 40 to 52. Tissue samples for immunohistochemical analysis were obtained at autopsy, frozen in liquid nitrogen, and stored at -80°C or were fixed in 10% buffered formalin and processed for paraffin section. The pathological features of these cases were also typical for SBMA,^2,3 with minimal variation in extent among the patients; the spinal, bulbar and pontine motor neurons were extensively depleted, with mild gliosis; sensory neurons were mildly affected, with occasional Nageotte's nodules; the posterior column of the spinal cord was depleted in a rostrally accentuated manner; the muscles were chronically denervated, and the sural nerve myelinated fibers were moderately depleted. Testicular atrophy and fatty liver changes were also present. Other portions of the central nervous system and visceral organs were normal except for pulmonary infection in all patients and gastric cancer in one patient.

Control tissue samples were obtained from four male autopsied patients ages 54 to 71 years, who died of nonneurological diseases. The AR CAG repeat lengths of the controls were 19 to 24.

All autopsies were performed within 6 hours postmortem.

Abs to AR Protein Used in This Study

Several polyclonal and monoclonal Abs that specifically recognize the AR protein were used in this study (Figure 1) : 2F12 (mouse monoclonal Ab (immunoglobulin (Ig) G), NovoCastra, Newcastle, UK), generated against a recombinant protein of 321 amino acids from the N terminus of the human AR; PG-21 (rabbit polyclonal Ab (IgG), Affinity BioReagents, Golden, CO) and AR(N-20)(rabbit polyclonal Ab (IgG), Santa Cruz Biotechnology, Santa Cruz, CA), which recognize 21 and 20 amino acid residues of the N terminus of the AR, respectively; AR52 (rabbit polyclonal Ab (IgG), kindly provided by Dr. E. Wilson, University of North Carolina, Chapel Hill, NC), which recognizes the DNA-binding domain of the AR; and 5F4 (mouse monoclonal Ab (IgM), kindly provided by Dr. T Demura, Department of Urology, Hokkaido University, Hokkaido, Japan) and AR(C-19) (rabbit polyclonal Ab (IgG), Santa Cruz Biotechnology), which recognizes the C terminus of the human AR. Characterization and binding specificities of all of these Abs to human AR were previously described.^33,37-39 Anti-ubiquitin Ab (rabbit polyclonal Ab (IgG), Dakopatts, Glostrup, Denmark) was also used.

View larger version (11K): [in this window] [in a new window]   Figure 1. Locations of the epitopes of Abs in the human AR.

Cryostat sections of 8 µm were prepared from the frozen tissues of SBMA patients and controls, quickly dried, and lightly fixed with Zamboni fixative for 10 minutes. Then the tissue sections were washed, blocked with normal horse serum (1:20), and incubated with Abs against AR, ubiquitin, or affinity-purified mouse IgG1 at concentrations of 0.5 to 4 µg/ml. Endogenous peroxidase was blocked by preincubation of tissue sections with 0.3% H₂O₂ in methanol for 30 minutes. Endogenous biotin was also blocked by incubation with an avidin-biotin blocking kit (Vector Laboratories, Burlingame, CA). Immune complexes were visualized using the avidin-biotinylated horseradish peroxidase system (Elite Vector kit from Vector Laboratories) and 3,3'-diaminobenzidine (Dakopatts) substrate. Sections were counterstained with methyl green. For paraffin-embedded samples, the 4-µm tissue sections were deparaffinized in xylene, hydrated with alcohol, and then heated in a microwave oven for 5 minutes. The tissue sections were then processed in the same way as those for the frozen tissue sections. Immune complexes were visualized using the tyramide signal amplification system (New England Nuclear, Boston, MA) following the manufacturer's protocol.

For electron microscopic immunohistochemistry, buffered formalin-fixed, paraffin-embedded tissue sections were immunostained with Abs against AR and then incubated with horseradish peroxidase-labeled second Ab (Amersham, Poole, UK). The tissue sections were then visualized with 3,3'-diaminobenzidine (Dakopatts), fixed with 2% osmium tetroxide in 0.1 mol/L phosphate buffer, pH 7.4, and dehydrated in an alcohol gradient and embedded in epoxy resin, from which ultrathin sections were obtained and then observed under an electron microscope (Hitachi H-7000).

CAG Repeat Length Assessment for Normal and Mutant AR Genes

CAG repeat length of the AR gene was determined on the autopsied tissue samples using methods described previously.^5,40

	Results

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Nuclear inclusions were detected in the spinal motor neurons, neurons in the motor nucleus of the trigeminal nuclei in the pons, and neurons of the hypoglossal nuclei of the medulla oblongata (Figure 2 (see also Figure 4 ), Table 1 ). Nuclear inclusions were not detected in the neurons of cerebral cortex, caudate, thalamus, pallidum, cerebellar cortex, dentate nucleus nuclei other than motor nuclei in the brain stem, intermediolateral and Clarke's nuclei of the spinal cord, or dorsal root ganglion neurons. Nuclear inclusions similar to those in the motor neurons were detected in the scrotal skin, dermal skin, kidney, heart, and testis (Figures 3 and 4 , Table 1 ). The inclusions were not seen in the liver, spleen, or muscle. Both neural and nonneural nuclear inclusions were strongly stained by PG-21 and AR(N-20), but not by 2F12, AR52, 5F4, or AR(C-19) (Table 2) . The inclusions were spherical structures of 1 to 5 µm in diameter, and those in the nonneural tissues were generally smaller in size than those in the neurons. They were ubiquitinated (Figure 3 , Tables 1 and 2 ). There were no detectable inclusions in cytoplasm of the neural or nonneural tissues. Most commonly we saw one inclusion in the nucleus of the individual cells, but two or three inclusions per cell were also observed (Figures 2 to 4) in both the neural and nonneural tissues. The frequency of the nuclear inclusions in the spinal motor neurons was 8.39 ± 5.43% of total remaining motor neurons. The frequencies of nuclear inclusions in nonneural tissues were 0.93 ± 0.50% in scrotal skin epidermal cells, 0.40% in nonscrotal skin epidermal cells, and 0.59 ± 0.02% in kidney tubular cells, and nuclear inclusions were observed only occasionally in the heart and testis.

View larger version (159K): [in this window] [in a new window]   Figure 2. Immunohistochemistry of the AR protein in nuclear inclusions in the central nervous system of SBMA patients. AR staining is observed in motor neurons of the medulla oblongata (A and B) and pons (C and D). A and B are stained with PG-21, and C and D are stained with AR(N-20). Magnification: A to D, 600x.

View larger version (150K): [in this window] [in a new window]   Figure 4. Electron microscopic immunohistochemical features of the nuclear inclusions in motor neurons and scrotal skin epidermal cells of SBMA patients. Dense granular aggregation of AR-immunoreactive material without a limiting membrane was found in both motor neurons (A to C) and scrotal skin epidermal cells (D to F). Nucleoli, which are larger than the inclusions, are clearly distinguished. The same inclusions observed light microscopically (A and D) are demonstrated electron microscopically (B, C, E, and F). AR(N-20) was used as the first Ab. Magnification: A, 720x; D, 1,000x; B and E, 2,000x; C, 10,000x; F, 15,000x.

View larger version (144K): [in this window] [in a new window]   Figure 3. Nuclear inclusions in nonneural tissues of SBMA patients. AR staining is observed in epithelial cells in the proximal tubules of the kidney (A), scrotal skin epidermal cells (C), and testis (E). PG-21 was used as the first Ab. These inclusions all stained positively for ubiquitin (B, D and F). Magnification: A to F, 840x.

The morphological appearance of the motor neurons and nonneural cells with nuclear inclusions was indistinguishable from those without inclusions.

Neural and nonneural tissues from four control cases were also examined in the same manner as that for SBMA cases, but the inclusions were not seen in the control individuals.

	Discussion

Top Abstract Introduction Materials and Methods Results Discussion References

 
The present study demonstrates that nuclear inclusions of the mutant AR protein are present in the motor neurons of the spinal cord and the brain stem; moreover, similar inclusions are also detected in certain nonneural tissues of the SBMA patients, indicating that the occurrence of the nuclear inclusions is not restricted to the affected neural tissues, but is also found in certain nonneural tissues.

The electron microscopic appearance of the AR-positive dense aggregates with similar granular size without limiting membrane was common to both neural and nonneural tissues. Furthermore, a characteristic selective staining pattern of the nuclear inclusions seen with only Abs recognizing N-terminal 20 and 21 amino acids of the AR protein was also common among the neural and nonneural tissues. These observations strongly suggest that same mechanism is involved in formation of AR-positive components in the nuclear inclusions in both the neural and nonneural tissues. The immunostaining pattern suggests that only a small portion of the N terminus of the AR protein is available as an epitope in the nuclear inclusions, and other portions of the AR may be masked within the inclusions of the mutant protein, or alternatively, the AR protein may be cleaved by proteolytic activity resulting in N-terminal fragments that participate in the aggregation, as was suggested in other polyglutamine diseases.^26-36,41 In addition, these AR nuclear inclusions are ubiquitinated in the nonneural as well as neural tissues, indicating that the nuclear inclusion is a pathological structure of the mutant AR, even in the nonneural tissues, where the pathological involvement is not apparent. Our electron microscopic and light microscopic immunohistochemical data indicate that nuclear inclusions in both motor neurons and nonneural tissues are identical in morphological and immunochemical features. Furthermore, absence of filamentous structures in the nuclear inclusion of SBMA was different from observations in HD²⁷ and MJD.³⁰ This difference may suggest that the pathway of the nuclear aggregation is different among the different protein products or, alternatively, may represent variances in sample preparation.

In polyglutamine diseases that have been analyzed to date, the nuclear inclusions have been shown to occur selectively in neurons of the affected brain regions. The selective occurrence of nuclear inclusions in the affected cells of central nervous system in SBMA that we observed in this study agrees well with observations of HD, MJD, SCA1, and DRPLA,^27-32 as well as our previous observations in SBMA.³³ However, the appearance of similar nuclear inclusions in the nonneural tissues observed in this study is novel. It is an important question why the neurons are selectively affected despite the presence of nuclear inclusions in both the affected neurons and nonaffected nonneural tissues. The cells of the nonneural tissues are mitotic cells in contrast to motor neurons; the epidermal cells in the scrotal and dermal skin and epithelial cells in the kidney tubules are all capable of mitosis in adulthood, and are eventually replaced by newly generated cells. Hence, those cells with toxic effects associated with the nuclear inclusions may be replaced by turnover. It may also be that neurons are particularly susceptible to whatever deleterious effects the inclusions may have. As demonstrated in the HD transgenic model,³⁴ a long latent period is necessary for inclusions to induce neuronal death. Neurons, as postmitotic cells, may be specifically affected because they survive long enough for the inclusions to have effect, whereas nonneural cells with nuclear inclusions turn over before the inclusions have pathological consequences. The significantly lower frequency of nuclear inclusion in nonneural tissues than in neurons may support this view. These differences in cell turnover rates could contribute to selective neuronal degeneration and neuronal loss.

Another interesting observation in this study is that the presence of nuclear inclusions is also selective among the various nonneural tissues, as it is in neural tissues. The inclusions are frequent in scrotal skin, dermal skin, and kidney; only occasionally seen in the testis and heart muscle; and not detected in spleen, liver, and muscle. The distribution of inclusions is not related to the expression level of mutant AR in these tissues.³³ AR protein is highly expressed in the testis, skin, and muscle, whereas it is low in the kidney, spleen, and liver.³³ A similar lack of correlation between the AR protein expression levels and pathological involvement is seen in the nervous system; neurons in Onuf's nuclei and Purkinje cells, for example, express a relatively large amount of AR protein,³³ but these neurons are not affected in SBMA. The underlying mechanism that induces the selective formation of nuclear inclusions in neural as well as nonneural tissues remains undetermined; specific factors may be present only in the tissues with the nuclear inclusions, such as a protease that cleaves the N-terminal portion from the full-length AR protein, or specific proteins that interact with the mutant AR, as demonstrated in SCA1^23,26 and HD,^21,22,24,25 may contribute to the cell type-specific formation of the nuclear inclusions. Alternatively, a degenerative process leading to apoptotic cell death with activation of a protease such as caspase 3 may be involved in the formation of nuclear inclusions as proposed in HD, DRPLA, MJD, and SBMA.^41,42 Furthermore, the tissue-specific inclusion formation could be influenced by the different AR functional activity among the different tissues. The nonneural tissues we found to contain nuclear inclusions have relatively rapid cell turnover, with physiological cell degeneration and cell death.

In summary, cell type-specific nuclear inclusions are present in the nonneural as well as neural tissues in SBMA, but the high physiological rate of cell turnover may contribute to the absence of functional impairment with the formation of nuclear inclusions in the nonneural tissues.

	Footnotes

 
Address reprint requests to Dr. Gen Sobue, Department of Neurology, Nagoya University School of Medicine, Tsurumai-cho 65, Showa-ku, Nagoya 466, Japan. E-mail: sobueg{at}tsuru.med.nagoya-u.ac.jp

This work was supported by grants from the Ministry of Health and Welfare of Japan, the Muscular Dystrophy Association, the National Institutes of Health, and the Kanae Foundation for Life and Medical Science, and by a Center of Excellence grant from the Ministry of Education, Science and Culture of Japan.

Accepted for publication June 25, 1998.

	References

Top Abstract Introduction Materials and Methods Results Discussion References

 

1. Kennedy WR, Alter M, Sung JH: Progressive proximal spinal and bulbar muscular atrophy of late onset: a sex-linked recessive trait. Neurology 1968, 18:671-680[Free Full Text]
2. Sobue G, Hashizume Y, Mukai E, Hirayama M, Mitsuma T: X-linked recessive bulbospinal neuronopathy, a clinicopathological study. Brain 1989, 112:209-232[Abstract/Free Full Text]
3. Li M, Sobue G, Doyu M, Mukai E, Hashizume Y, Mitsuma T: Primary sensory neurons in X-linked recessive bulbospinal neuronopathy: histopathology and androgen receptor gene expression. Muscle Nerve 1995, 18:301-308[Medline]
4. La Spada AR, Wilson EM, Lubahn DB, Harding AE, Fischbeck KH: Androgen receptor gene mutations in X-linked spinal and bulbar muscular atrophy. Nature 1991, 352:77-79[Medline]
5. Doyu M, Sobue G, Mukai E, Kachi T, Yasuda T, Mitsuma T, Takahasi A: Severity of X-linked recessive bulbospinal neuronopathy correlates with size of the tandem CAG repeat in androgen receptor gene. Ann Neurol 1992, 32:707-710[Medline]
6. Igarashi S, Tanno Y, Onodera O, Yamazaki M, Sato S, Ishikawa A, Miyatani N, Nagashima M, Ishikawa Y, Sahashi K, Ibi T, Miyatake T, Tsuji S: Strong correlation between the number of CAG repeats in androgen receptor genes and the clinical onset of features of spinal and bulbar muscular atrophy. Neurology 1992, 42:2300-2302[Abstract/Free Full Text]
7. La Spada AR, Roling DB, Harding AE, Warner CL, Spiegel R, Hausmanowa-Petrusewicz I, Yee WC, Fischbeck KH: Meiotic stability and genotype-phenotype correlation of the trinucleotide repeat in X-linked spinal and bulbar muscular atrophy. Nat Genet 1992, 2:301-304[Medline]
8. Shimada N, Sobue G, Doyu M, Yamamoto K, Yasuda T, Mukai E, Kachi T, Mitsuma T: X-linked recessive bulbospinal neuronopathy: clinical phenotypes and CAG repeat size in androgen receptor gene. Muscle Nerve 1995, 18:1378-1384[Medline]
9. : The Huntington's Disease Collaborative Research Group: A novel gene containing a trinucleotide repeat that is expanded and unstable on Huntington's disease chromosomes. Cell 1993, 72:971-983[Medline]
10. Koide R, Ikeuchi T, Onodera O, Tanaka H, Igarashi S, Endo K, Takahashi H, Kondo R, Ishikawa A, Hayashi T, Saito M, Tomoda A, Miike T, Naito H, Ikuta F, Tsuji S: Unstable expansion of CAG repeat in hereditary dentatorubral-pallidoluysian atrophy (DRPLA). Nat Genet 1994, 6:9-13[Medline]
11. Nagafuchi S, Yanagisawa H, Sato K, Shirayama T, Ohsaki E, Bundo M, Takeda T, Tadokoro K, Kondo I, Murayama N, Tanaka Y, Kikushima H, Umino K, Kurosawa H, Furukawa T, Nihei K, Inoue T, Sano A, Komura O, Takahashi M, Yoshizawa T, Kanazawa I, Yamada M: Dentatorubral and pallidoluysian atrophy: expansion of an unstable CAG trinucleotide on chromosome 12p. Nat Genet 1994, 6:14-18[Medline]
12. Kawaguchi Y, Okamoto T, Taniwaki M, Aizawa M, Inoue M, Katayama S, Kawakami H, Nakamura S, Nisimura M, Akiguchi I, Kimura J, Narumiya S, Kakizuka A: CAG expansion in a novel gene from Machado-Joseph disease at chromosome 14q32.1. Nat Genet 1994, 8:221-227[Medline]
13. Orr HT, Cung M-Y, Banfi S, Kwiatkowski JTJ, Servadio A, Beaudet AL, McCall AE, Duvick LA, Ranum LPW, Zoghbi HY: Expansion of an unstable trinucleotide CAG repeat in spinocerebellar ataxia type 1. Nat Genet 1993, 4:221-226[Medline]
14. Pulst SM, Nechiporuk A, Nechiporuk T, Gispert S, Chen XN, Cendes IL, Pearlman S, Starkman S, Diaz GO, Lunkes A, DeJong P, Rouleau GA, Auburger G, Korenberg JR, Figueroa C, Sahba S: Moderate expansion of a normally biallelic trinucleotide repeat in spinocerebellar ataxia type 2. Nat Genet 1996, 14:269-276[Medline]
15. Sanpei K, Takano H, Igarashi S, Sato T, Oyake M, Sasaki H, Wakisaka A, Tashiro K, Ishida Y, Ikeuchi T, Koide R, Saito M, Sato A, Tanaka T, Hanyu S, Takiyama Y, Nishizawa M, Shimizu N, Nomura Y, Segawa M, Iwabuchi K, Eguchi I, Tanaka H, Takahashi H, Tsuji S: Identification of the spinocerebellar ataxia type-2 gene using a direct identification of repeat expansion and cloning technique, DIRECT. Nat Genet 1996, 14:277-284[Medline]
16. Imbert G, Saudou F, Yvert G, Devys D, Trottier Y, Garnier JM, Weber C, Mandel JL, Cancel G, Abbas N, Dürr A, Didierjean O, Stevanin G, Agid Y, Brice A: Cloning of the gene for spinocerebellar ataxia type 2 reveals a locus with high sensitivity to expanded CAG/glutamine repeats. Nat Genet 1996, 14:285-291[Medline]
17. Zhuchenko O, Bailey J, Bonnen P, Ashizawa T, Stockton DW, Amos C, Dobyns WB, Subramony SH, Zoghbi HY, Lee CC: Autosomal dominant cerebellar ataxia (SCA 6) associated with small polyglutamine expansion in the a_1A-voltage-dependent calcium channel. Nat Genet 1997, 15:62-69[Medline]
18. David G, Abbas N, Stevanin G, Durr A, Yvert G, Cancel G, Weber C, Imbert G, Saudou F, Antoniou E, Drabkin H, Gemmill R, Giunti P, Benonar A, Wood N, Ruberg M, Agid Y, Mandel JL, Brice A: Cloning of the SCA 7 gene reveals a highly unstable CAG repeat expansion. Nat Genet 1997, 17:65-70[Medline]
19. Perutz MF, Johnson T, Suzuki M, Finch JT: Glutamine repeats as polar zippers: their possible role in inherited neurodegenerative diseases. Proc Natl Acad Sci USA 1994, 91:5355-5358[Abstract/Free Full Text]
20. Li X-J, Li S-H, Sharp AH, Nucifora JFC, Schilling G, Lanahan A, Worley P, Snyder SH, Ross CA: A huntingtin-associated protein enriched in brain with implication for pathology. Nature 1995, 378:398-402[Medline]
21. Ikeda H, Yamaguchi M, Sugai S, Aze Y, Narumiya S, Kakizuka A: Expanded polyglutamine in the Machado-Joseph disease protein induces cell death in vitro and in vivo. Nat Genet 1996, 13:196-202[Medline]
22. Burke JR, Enghild JJ, Martin ME, Jou YS, Myers RM, Roses AD, Vance JM, Strittmatter WJ: Huntingtin and DRPLA proteins selectively interact with the enzyme GAPDH. Nat Med 1996, 2:347-349[Medline]
23. Koshy B, Matilla T, Merry DE, Fischbeck KH, Orr HT, Zoghbi HY: Spinocerebellar ataxia type-1 and spinobulbar muscular atrophy gene products interact with glyceraldehyde-3-phosphate dehydrogenase. Hum Mol Genet 1996, 5:1311-1318[Abstract/Free Full Text]
24. Kalchman MA, Koide HB, McCutcheon K, Graham RK, Nichol K, Nishiyama K, Kazemi-Esfarjani P, Lynn FC, Wellington C, Metzler M, Goldberg YP, Kanazawa I, Gietz RD, Hayden MR: HIP1, a human homologue of S. cerevisiae sla2p, interacts with membrane-associated huntingtin in the brain. Nat Genet 1997, 16:44-53[Medline]
25. Wanker EE, Rovira C, Scherzinger E, Hasenbank R, Walter S, Tait D, Colicelli J, Lehrach H: HIP-1: a huntingtin interacting protein isolated by the yeast two hybrid system. Hum Mol Genet 1997, 6:487-495[Abstract/Free Full Text]
26. Matilla A, Koshy BT, Cummings CJ, Isobe T, Orr HT, Zoghbi HY: The cerebellar leucine-rich acidic nuclear protein interacts with ataxin-1. Nature 1997, 389:974-978[Medline]
27. DiFiglia M, Sapp E, Chase KO, Davies SW, Bates GP, Vonsattel JP, Aronin N: Aggregation of huntingtin in neuronal intranuclear inclusions and dystrophic neurites in brain. Science 1997, 227:1990-1993
28. Martindale D, Hackam A, Wieczorek A, Ellerby L, Wellington C, McCutcheon K, Singaraja R, Kazemi-Esfarjani P, Devon R, Kim SU, Bredesen DE, Tufaro F, Hayden MR: Length of huntingtin and its polyglutamine tract influences localization and frequency of intracellular aggregates. Nat Genet 1998, 18:150-154[Medline]
29. Paulson HL, Das SS, Crino PB, Perez MK, Patel SC, Gotsdiner D, Fischbeck KH, Pittman RN: Machado-Joseph disease gene product is a cytoplasmic protein expressed widely in brain. Ann Neurol 1997, 41:453-462[Medline]
30. Paulson HL, Perez DMK, Troffier Y, Trojanowski JQ, Subramony SH, Das SS, Vig P, Mandel J-L, Fischbeck KH, Pittman RN: Intranuclear inclusions of expanded polyglutamine protein in spinocerebellar ataxia type 3. Neuron 1997, 19:333-344[Medline]
31. Skinner PJ, Koshy BT, Cummings CJ, Klement IA, Helin K, Servadio A, Zoghbi H, Orr HT: Ataxin-1 with an expanded glutamine tract alters nuclear matrix-associated structures. Nature 1997, 389:971-974[Medline]
32. Igarashi S, Koike R, Shimohata T, Yamada M, Hayashi Y, Takano H, Date H, Oyake M, Sato T, Sato A, Egawa S, Ikeuchi T, Tanaka H, Nakano R, Tanaka K, Hozumi I, Inuzuka T, Takahashi H, Tsuji S: Suppression of aggregate formation and apoptosis by transglutaminase inhibitors in cells expressing truncated DRPLA protein with an expanded polyglutamine stretch. Nat Genet 1998, 18:111-117[Medline]
33. Li M, Kobayashi Y, Merry D, Tanaka F, Doyu M, Hashizume Y, Fischbeck KH, Sobue G: Nuclear inclusions in spinal and bulbar muscular atrophy. Ann Neurol 1998 (in press)
34. Davies SW, Turmaine M, Cozens BA, DiFiglia M, Sharp AH, Ross CA, Scherzinger E, Wanker EE, Mangiarini L, Bates GP: Formation of neuronal intranuclear inclusions underlies the neurological dysfunction in mice transgenic for the HD mutation. Cell 1997, 90:537-548[Medline]
35. Scherzinger E, Lurz R, Tumine M, Mangiarini L, Hollenbach B, Hasenbank R, Bates GP, Davies SW, Lehrach H, Wanker EE: Huntingtin-encoded polyglutamine expansions from amyloid-like protein aggregates in vitro and in vivo. Cell 1997, 90:549-558[Medline]
36. Ross CA: Intranuclear neuronal inclusions: a common pathogenic mechanism for glutamine-repeat neurodegenerative diseases? Neuron 1997, 19:1147-1150[Medline]
37. Prins GS, Birch L, Greene GL: Androgen receptor localization in different cell types of the adult rat prostate. Endocrinology 1991, 129:3187-3199[Abstract]
38. Sar M, Lubahn DB, French FS, Wilson EM: Immunohistochemical localization of the androgen receptor in rat and human tissues. Endocrinology 1990, 127:3180-3186[Abstract]
39. Demura T, Kuzumaki N, Oda A, Fujita H, Ishibashi T, Koyanagi T: Establishment and characterization of monoclonal antibody against androgen receptor. J Steroid Biochem 1987, 33:845-851
40. Tanaka F, Doyu M, Ito Y, Matsumoto M, Mitsuma T, Abe K, Aoki M, Itoyama Y, Fischbeck KH, Sobue G: Founder effect in spinal and bulbar muscular atrophy (SBMA). Hum Mol Genet 1996, 5:1253-1257[Abstract/Free Full Text]
41. Goldberg YP, Nicholson DW, Rasper DM, Kalchman MA, Koide HB, Graham RK, Bromm M, Kazemi-Esfarjani P, Thornberry NA, Vaillancourt JP, Hayden MR: Cleavage of huntingtin by apopain, a proapoptotic cysteine protease, is modulated by the polyglutamine tract. Nat Genet 1996, 13:442-449[Medline]
42. Wellington CL, Ellerby LM, Hackam AS, Margolis RL, Trifiro MA, Singaraja R, McCutcheon K, Salvesen GS, Propp SS, Bromm M, Rowland KJ, Zhang T, Rasper D, Roy S, Thornberry N, Pinsky L, Kakizuka A, Ross CA, Nicholson DW, Bredesen DE, Hayden MR: Caspase cleavage of gene products associated with triplet expansion disorders generates truncated fragments containing the polyglutamine tract. J Biol Chem 1998, 273:9158-9167[Abstract/Free Full Text]

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				N. Atsuta, H. Watanabe, M. Ito, H. Banno, K. Suzuki, M. Katsuno, F. Tanaka, A. Tamakoshi, and G. Sobue Natural history of spinal and bulbar muscular atrophy (SBMA): a study of 223 Japanese patients Brain, June 1, 2006; 129(6): 1446 - 1455. [Abstract] [Full Text] [PDF]

				H. Adachi, M. Katsuno, M. Minamiyama, M. Waza, C. Sang, Y. Nakagomi, Y. Kobayashi, F. Tanaka, M. Doyu, A. Inukai, et al. Widespread nuclear and cytoplasmic accumulation of mutant androgen receptor in SBMA patients Brain, March 1, 2005; 128(3): 659 - 670. [Abstract] [Full Text] [PDF]

				C. M. Everett and N. W. Wood Trinucleotide repeats and neurodegenerative disease Brain, November 1, 2004; 127(11): 2385 - 2405. [Abstract] [Full Text] [PDF]

				M. Minamiyama, M. Katsuno, H. Adachi, M. Waza, C. Sang, Y. Kobayashi, F. Tanaka, M. Doyu, A. Inukai, and G. Sobue Sodium butyrate ameliorates phenotypic expression in a transgenic mouse model of spinal and bulbar muscular atrophy Hum. Mol. Genet., June 1, 2004; 13(11): 1183 - 1192. [Abstract] [Full Text] [PDF]

				E. S. Chevalier-Larsen, C. J. O'Brien, H. Wang, S. C. Jenkins, L. Holder, A. P. Lieberman, and D. E. Merry Castration Restores Function and Neurofilament Alterations of Aged Symptomatic Males in a Transgenic Mouse Model of Spinal and Bulbar Muscular Atrophy J. Neurosci., May 19, 2004; 24(20): 4778 - 4786. [Abstract] [Full Text] [PDF]

				N. Hishikawa, J.-i. Niwa, M. Doyu, T. Ito, S. Ishigaki, Y. Hashizume, and G. Sobue Dorfin Localizes to the Ubiquitylated Inclusions in Parkinson's Disease, Dementia with Lewy Bodies, Multiple System Atrophy, and Amyotrophic Lateral Sclerosis Am. J. Pathol., August 1, 2003; 163(2): 609 - 619. [Abstract] [Full Text] [PDF]

				K. Ishihara, N. Yamagishi, Y. Saito, H. Adachi, Y. Kobayashi, G. Sobue, K. Ohtsuka, and T. Hatayama Hsp105{alpha} Suppresses the Aggregation of Truncated Androgen Receptor with Expanded CAG Repeats and Cell Toxicity J. Biol. Chem., June 27, 2003; 278(27): 25143 - 25150. [Abstract] [Full Text] [PDF]

				H. Adachi, M. Katsuno, M. Minamiyama, C. Sang, G. Pagoulatos, C. Angelidis, M. Kusakabe, A. Yoshiki, Y. Kobayashi, M. Doyu, et al. Heat Shock Protein 70 Chaperone Overexpression Ameliorates Phenotypes of the Spinal and Bulbar Muscular Atrophy Transgenic Mouse Model by Reducing Nuclear-Localized Mutant Androgen Receptor Protein J. Neurosci., March 15, 2003; 23(6): 2203 - 2211. [Abstract] [Full Text] [PDF]

				J. L. Walcott and D. E. Merry Ligand Promotes Intranuclear Inclusions in a Novel Cell Model of Spinal and Bulbar Muscular Atrophy J. Biol. Chem., December 20, 2002; 277(52): 50855 - 50859. [Abstract] [Full Text] [PDF]

				P. McManamny, H. S. Chy, D. I. Finkelstein, R. G. Craythorn, P. J. Crack, I. Kola, S. S. Cheema, M. K. Horne, N. G. Wreford, M. K. O'Bryan, et al. A mouse model of spinal and bulbar muscular atrophy Hum. Mol. Genet., September 1, 2002; 11(18): 2103 - 2111. [Abstract] [Full Text] [PDF]

				N. J. Caplen, J. P. Taylor, V. S. Statham, F. Tanaka, A. Fire, and R. A. Morgan Rescue of polyglutamine-mediated cytotoxicity by double-stranded RNA-mediated RNA interference Hum. Mol. Genet., January 1, 2002; 11(2): 175 - 184. [Abstract] [Full Text] [PDF]

				K. Sathasivam, B. Woodman, A. Mahal, F. Bertaux, E. E. Wanker, D. T. Shima, and G. P. Bates Centrosome disorganization in fibroblast cultures derived from R6/2 Huntington's disease (HD) transgenic mice and HD patients Hum. Mol. Genet., October 1, 2001; 10(21): 2425 - 2435. [Abstract] [Full Text] [PDF]

				H. Adachi, A. Kume, M. Li, Y. Nakagomi, H. Niwa, J. Do, C. Sang, Y. Kobayashi, M. Doyu, and G. Sobue Transgenic mice with an expanded CAG repeat controlled by the human AR promoter show polyglutamine nuclear inclusions and neuronal dysfunction without neuronal cell death Hum. Mol. Genet., May 1, 2001; 10(10): 1039 - 1048. [Abstract] [Full Text] [PDF]

				A. Abel, J. Walcott, J. Woods, J. Duda, and D. E. Merry Expression of expanded repeat androgen receptor produces neurologic disease in transgenic mice Hum. Mol. Genet., January 1, 2001; 10(2): 107 - 116. [Abstract] [Full Text] [PDF]

				A. McCampbell, J. P. Taylor, A. A. Taye, J. Robitschek, M. Li, J. Walcott, D. Merry, Y. Chai, H. Paulson, G. Sobue, et al. CREB-binding protein sequestration by expanded polyglutamine Hum. Mol. Genet., September 1, 2000; 9(14): 2197 - 2202. [Abstract] [Full Text] [PDF]

				M. Becker, E. Martin, J. Schneikert, H. F. Krug, and A. C.B. Cato Cytoplasmic Localization and the Choice of Ligand Determine Aggregate Formation by Androgen Receptor with Amplified Polyglutamine Stretch J. Cell Biol., April 17, 2000; 149(2): 255 - 262. [Abstract] [Full Text] [PDF]

				C. J. Cummings and H. Y. Zoghbi Fourteen and counting: unraveling trinucleotide repeat diseases Hum. Mol. Genet., April 1, 2000; 9(6): 909 - 916. [Abstract] [Full Text] [PDF]

				Y. Kobayashi, A. Kume, M. Li, M. Doyu, M. Hata, K. Ohtsuka, and G. Sobue Chaperones Hsp70 and Hsp40 Suppress Aggregate Formation and Apoptosis in Cultured Neuronal Cells Expressing Truncated Androgen Receptor Protein with Expanded Polyglutamine Tract J. Biol. Chem., March 17, 2000; 275(12): 8772 - 8778. [Abstract] [Full Text] [PDF]

				S. Simeoni, M. A. Mancini, D. L. Stenoien, M. Marcelli, N. L. Weigel, M. Zanisi, L. Martini, and A. Poletti Motoneuronal cell death is not correlated with aggregate formation of androgen receptors containing an elongated polyglutamine tract Hum. Mol. Genet., January 1, 2000; 9(1): 133 - 144. [Abstract] [Full Text] [PDF]

				L. V. Nazareth, D. L. Stenoien, W. E. Bingman III, A. J. James, C. Wu, Y. Zhang, D. P. Edwards, M. Mancini, M. Marcelli, D. J. Lamb, et al. A C619Y Mutation in the Human Androgen Receptor Causes Inactivation and Mislocalization of the Receptor with Concomitant Sequestration of SRC-1 (Steroid Receptor Coactivator 1) Mol. Endocrinol., December 1, 1999; 13(12): 2065 - 2075. [Abstract] [Full Text]

				M. D. Kaytor, L. A. Duvick, P. J. Skinner, M. D. Koob, L. P. W. Ranum, and H. T. Orr Nuclear localization of the spinocerebellar ataxia type 7 protein, ataxin-7 Hum. Mol. Genet., September 1, 1999; 8(9): 1657 - 1664. [Abstract] [Full Text] [PDF]

				H. Li, S.-H. Li, A. L. Cheng, L. Mangiarini, G. P. Bates, and X.-J. Li Ultrastructural localization and progressive formation of neuropil aggregates in Huntington's disease transgenic mice Hum. Mol. Genet., July 1, 1999; 8(7): 1227 - 1236. [Abstract] [Full Text] [PDF]

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