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Altered Expression of Carbonic Anhydrase-Related Protein XI in Neuronal Cells Expressing Mutant Ataxin-3

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Abstract

Spinocerebellar ataxia type 3/Machado–Joseph disease (SCA3/MJD) is a late-onset neurodegenerative disorder caused by the expansion of a polyglutamine tract within the gene product, ataxin-3. Microarray analysis revealed a dramatic differential expression of carbonic anhydrase-related protein XI (CA-RPXI/CA11) in the presence or absence of mutant ataxin-3. Therefore, we examined the expression and distribution of all three CA-RPs (CA8, 10, and 11) in human neuronal cells that stably express mutant ataxin-3. Compared with the cells containing normal ataxin-3, protein expression of CA8 and CA11 is significantly increased in human neuroblastoma cells harboring mutant ataxin-3. Semi-quantitative RT-PCR demonstrated that all three CA-RPs exhibited significantly higher transcript levels in neuronal cells expressing mutant ataxin-3. Interestingly, CA11 is distributed not only in the cytoplasm but also within the nuclei of the stably transfected mutant cells when compared with the sole cytoplasmic distribution in cells containing normal ataxin-3. In addition, results from transient transfection assays in SK-N-SH and Neuro2a (N2a) cells also confirmed the nuclear localization of CA11 in the presence of truncated ataxin-3. Most importantly, immunohistochemical staining of the MJD transgenic mouse and post-mortem MJD human brain also revealed that CA11 is highly expressed in both cytoplasm and nuclei of the brain cells. Recruitment of CA11 into nuclear inclusions containing mutant ataxin-3 revealed a possible correlation between CA11 and disease progression. Although the exact function of CA-RPs is still undefined in the central nervous system, our findings suggest that CA-RPs, especially CA11, may play specific roles in the pathogenesis of Machado–Joseph disease.

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Acknowledgments

We thank Drs. Akira Kakizuka and Henry Paulson for MJD plasmids, Drs. Cemal Cemal and Yung-Hsiang Hsu for brain tissues, Dr. Shin-Lan Hsu for SK-N-SH cells, Dr. Chi-Min Shih for his assessment and interpretation of immunostaining data, Benjamin Hsieh for editing the manuscript, and Tang-Hao Chi and Tzu-Kai Wang for assistance of figure preparation. This work was supported by grants from the National Science Council of the Republic of China (NSC-91-2320-B-040-038; NSC-93-2311-B-029-009; NSC-97-2311-B-029-001-MY3).

Conflict of Interest

The authors declare that there is no conflict of interest with respect to financial or personal relationships with organizations that may have influence on this work.

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Authors and Affiliations

  1. Department of Life Science, Tunghai University, No.1727, Sec.4, Taiwan Boulevard, Taichung, Taiwan, 40704, Republic of China

    Mingli Hsieh, Wei-Hsiu Chang & Chi-Fu Hsu

  2. Life Science Research Center, Tunghai University, Taichung, Taiwan, 40704, Republic of China

    Mingli Hsieh

  3. Department of Gastroenterology, Kochi Medical School, Nankoku, Kochi, 783-8505, Japan

    Isao Nishimori & Tomoko Minakuchi

  4. Department of Biomedical Science, Chung-Shan Medical University, Taichung, Taiwan, 402, Republic of China

    Cheng-Liang Kuo

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  1. Mingli Hsieh
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Correspondence to Mingli Hsieh.

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Hsieh, M., Chang, WH., Hsu, CF. et al. Altered Expression of Carbonic Anhydrase-Related Protein XI in Neuronal Cells Expressing Mutant Ataxin-3. Cerebellum 12, 338–349 (2013). https://doi.org/10.1007/s12311-012-0430-2

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