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Increased T-type Ca2+ channel activity as a determinant of cellular toxicity in neuronal cell lines expressing polyglutamine-expanded human androgen receptors

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Abstract

We have analyzed Ca2+ currents in two neuroblastoma-motor neuron hybrid cell lines that expressed normal or glutamine-expanded human androgen receptors (polyGln-expanded AR) either transiently or stably. The cell lines express a unique, low-threshold, transient type of Ca2+ current that is not affected by L-type Ca2+ channel blocker (PN 200-110), N-type Ca2+ channel blocker (ω-conotoxin GVIA) or P-type Ca2+ channel blocker (Agatoxin IVA) but is blocked by either Cd2+ or Ni2+. This pharmacological profile most closely resembles that of T-type Ca2+ channels [1-3]. Exposure to androgen had no effect on control cell lines or cells transfected with normal AR but significantly changed the steady-state activation in cells transfected with expanded AR. The observed negative shift in steady-state activation results in a large increase in the T-type Ca2+ channel window current. We suggest that Ca2+ overload due to abnormal voltage-dependence of transient Ca2+ channel activation may contribute to motor neuron toxicity in spinobulbar muscular atrophy (SBMA). This hypothesis is supported by the additional finding that, at concentrations that selectively block T-type Ca2+ channel currents, Ni2+ significantly reduced cell death in cell lines transfected with polyGln-expanded AR.

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References

  1. Bkaily G, Sculptoreanu A, Jacques D, Jasmin G: Increases of T-type Ca2+ current in heart cells of cardiomyopathic hamster. Mol Cell Biochem 176: 199-204, 1997

    Google Scholar 

  2. Ertel SI, Ertel EA: Low-voltage-activated T-type Ca2+ channels. Trends Pharmacol Sci 18: 37-42, 1997

    Google Scholar 

  3. Nooney JM, Lambert MC, Feltz A: Identifying neuronal non-L Ca2+ channels-more than stamp collecting? Trends Pharmacol Sci 18: 363-371, 1997

    Google Scholar 

  4. Johansson J, Forsgren L, Sandgren O, Brice A, Holmgren G, Holmberg M: Expanded CAG repeats in Swedish spinocerebellar ataxia type 7 (SCA7) patients: Effect of CAG repeat length on the clinical manifestation. Hum Mol Genet 7: 171-176, 1998

    Google Scholar 

  5. Ross CA: Intranuclear neuronal inclusions: A common pathogenic mechanism for glutamine-repeat neurodegenerative diseases? Neuron 19: 1147-1150, 1997

    Google Scholar 

  6. Brooks BP, Fischbeck KH: Spinal and bulbar muscular atrophy: A trinucleotide-repeat expansion neurodegenerative disease. Trends Neurosci 10: 459-461, 1995

    Google Scholar 

  7. Brooks BP, Merry DE, Paulson HL, Lieberman AP, Kilson DL, Fischbeck KH: A cell culture model for androgen effects in motor neurons. J Neurochem 70: 1054-1060, 1998

    Google Scholar 

  8. Kennedy WR, Alter M, Sung JH: Progressive proximal spinal and bulbar muscular atrophy of late onset. A sex-linked recessive trait. Neurol 18: 671-680, 1968

    Google Scholar 

  9. Paulson HL, Fischbeck KN: Trinucleotides repeats in neurogenetic disorders. Annu Rev Neurosci 19: 79-100, 1996

    Google Scholar 

  10. La Spada AR, Wilson EM, Lubahn DB, Harding AB, Fischbeck KH: Androgen receptor gene mutations in X-linked spinal and bulbar muscular atrophy. Nature 352: 77-79, 1991

    Google Scholar 

  11. Matsuyama Z, Kawakami H, Maruyama H, Izumi Y, Komure O, Udaka F, Kameyama M, Nishio T, Kuroda Y, Nishimura M, Nakamura S: Molecular features of the CAG repeats of spinocerebellar ataxia 6 (SCA6). Hum Mol Genet 6: 1283-1287, 1997

    Google Scholar 

  12. Andrew SW, Goldberg YP, Hayden, MR: Rethinking genotype and phenotype correlations in polyglutamine expansion disorders. Hum Mol Genet 6: 2005-2010, 1997

    Google Scholar 

  13. Trifiro MA, Kazemi-Esfarjani P, Pinsky L: X-linked muscular atrophy and the androgen receptor. Trends Endocrinol Metab 5: 416-421, 1994

    Google Scholar 

  14. Nordeen EJ, Nordeen KW, Sengelaub DR, Arnold AP: Androgens prevent normally occurring cell death in a sexually dimorphic spinal nucleus. Science 229: 671-682, 1985

    Google Scholar 

  15. Perez J, Kelley DB: Trophic effects of androgen: Receptor expression and the survival of laryngeal motor neurons after axotomy. J Neurosci 16: 6625-6631, 1996

    Google Scholar 

  16. Perez J, Kelley DB: Androgen mitigates axotomy-induced decreases in calbindin expression in motor neurons. J Neurosci 17: 7396-7403, 1997

    Google Scholar 

  17. Tanzer L, Jones KJ: Gonadal steroid regulation of hamster facial nerve regeneration: Effects of dihydrotestosterone and estradiol. Exp Neurol 146: 258-266, 1997

    Google Scholar 

  18. Yu WH: Administration of testosterone attenuates neuronal loss following axotomy in the brain-stem motor nuclei of female rats. J Neurosci 9: 3908-3922, 1989

    Google Scholar 

  19. Zhuchenko O, Bailey J, Bonnen P, Ashizawa T, Stockton DW, Amos C, Dobyns WB, Subramony SH, Zoghbi HY, Lee CC: Autosomal cerebellar ataxia (SCA6) associated with small polyglutamine expansions in the a1A-voltage-dependent calcium channel. Nature Genet 15: 62-69, 1997

    Google Scholar 

  20. Butler R, Leigh PN, McPhaul MJ, Gallo JM: Truncated forms of the androgen receptor are associated with polyglutamine expansion in Xlinked spinal and bulbar muscular atrophy. Hum Mol Genet 7: 121-127, 1998

    Google Scholar 

  21. Abdullah AAR, Trifiro M.A, Panet-Raymond V, Alvarado C, de Tourreil S, Frankel D, Schipper HM, Pinsky L: Spinobulbar muscular atrophy: Polyglutamine-expanded androgen receptor is trypsin resistant in vitro and processed abnormally in transfected cells. Hum Mol Genet 7: 379-384, 1998

    Google Scholar 

  22. Wellington CL, Ellerby L M, 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 273: 9158-9167, 1998

    Google Scholar 

  23. Ellerby L, Hackman A, Propp S, Ellerby H, Rabizadeh S, Trifiro M, Pinsky L: Kennedy's disease: Caspase cleavage of the androgen receptor is a crucial event in cytotoxicity. J Neurochem 72: 185-195, 1998

    Google Scholar 

  24. Perutz MF: Glutamine repeats and inherited neurodegenerative diseases: molecular aspects. Curr Opin Struct Biol 6: 848-858, 1996

    Google Scholar 

  25. 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. Nature Genet 13: 196-202, 1996

    Google Scholar 

  26. Kahlem P, Terré C, Green H, Djian P: Peptides containing glutamine repeats as substrates for transglutaminase-catalyzed cross-linking: Relevance to diseases of the nervous system. Proc Natl Acad Sci USA 93: 14580-14585, 1996

    Google Scholar 

  27. Merry DE, Kobayashi Y, Bailey CK, Taye AA, Fischbeck KH: Cleavage, aggregation and toxicity of the expanded androgen receptor in spinal and bulbar muscular atrophy. Hum Mol Genet 7: 631-701, 1998

    Google Scholar 

  28. Paulson HL: Protein fate in neurodegenerative proteinpathies: Polyglutamine diseases join the (mis)fold. Am J Hum Genet 64: 339-345, 1999

    Google Scholar 

  29. Cashman N, Durham HD, Blusztajn JK, Oda K, Tabira T, Shaw IT, Dahrouge S, Antel JP: Neuroblastoma x spinal cord (NSC) hybrid cell lines resemble developing motor neurons. Dev Dynamics 194: 209-221, 1992

    Google Scholar 

  30. Salazar-Grueso EF, Kim S, Kim H: Embryonic mouse spinal cord motor neuron hybrid cells. Neuroreport 2: 505-508, 1991

    Google Scholar 

  31. Brooks BP, Paulson HL, Merry DE, Salazar-Grueso EF, Brinkmann AO, Wilson EM, Fischbeck KH: Characterization of an expanded glutamine repeat androgen receptor in a neuronal cell culture system. Neurobiol Dis 4: 313-323, 1997

    Google Scholar 

  32. Martin SJ, Matear PM, Vyakarnam A: HIV-1 infection of CD4 and T cells in vitro: Differential induction of apoptosis in these cells. J Immunol 152: 330-342, 1994

    Google Scholar 

  33. Sculptoreanu A, Figourov A, deGroat WC: Voltage-dependent potentiation of neuronal L-type Ca2+ channel currents due to statedependent phosphorylation. Am J Physiol 269: C725-C732, 1995

    Google Scholar 

  34. Bean BP, McDonough SI: Two for T. Neuron 20: 825-828, 1998

    Google Scholar 

  35. Bhattacharjee A, Whitehurst RM, Zhang M, Wang L, Li M: T-Type Calcium channels facilitate insulin secretion by enhancing general excitability in the insulin-secreting β-cell line, INS-1. Endocrinology 138: 3735-3740, 1997

    Google Scholar 

  36. McConkey DJ, Orrenius S: The role of calcium in the regulation of apoptosis. Biochem Biophys Res Commun 20: 357-366, 1997

    Google Scholar 

  37. Unemiya M, Araki I, Kuno M: Electrophysiological properties of axotomized facial motoneurones that are destined to die in neonatal rats. J Physiol (Lond) 462: 661-678, 1993

    Google Scholar 

  38. George EB, Glass JD, Griffin JW: Axotomy-induced axonal degeneration is mediated by calcium influx through ion-specific channels. J Neurosci 15: 6445-6452, 1995

    Google Scholar 

  39. Lesort M, Attanavanich K, Zhang J, Johnson GVW: Distinct nuclear localization and activity of tissue transglutaminase. J Biol Chem 273: 11991-11994, 1998

    Google Scholar 

  40. Igarashi S, Koide 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. Nature Genet 18: 111-117, 1998

    Google Scholar 

  41. Wang L. Bhattacharjee A. Zuo Z. Hu FQ. Honkanen RE. Berggren PO. Li M: A low voltage-activated Ca2+ current mediates cytokineinduced pancreatic beta-cell death. Endocrinology 140: 1200-1204, 1999

    Google Scholar 

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Authors and Affiliations

  1. Lady Davis Institute for Medical Research, SMBD-Jewish General Hospital, Canada

    Adrian Sculptoreanu, Abdullah A.R. Abdullah, Valerie Panet-Raymond, Dov Frankel, Hyman M. Schipper, Leonard Pinsky & Mark A. Trifiro

  2. Department of Experimental Medicine and Surgery, McGill University, Montreal, Quebec, H3G 1Y6, Canada

    Adrian Sculptoreanu

  3. Lady Davis Institute for Medical Research, SMBD-Jewish General Hospital, McGill University, Montreal, Quebec, H3G 1Y6, Canada

    Hanan Abramovici & Anna Bibikova

  4. Department of Biology, McGill University, Montreal, Quebec, H3G 1Y6, Canada

    Abdullah A.R. Abdullah & Valerie Panet-Raymond

  5. Department of Neurology and Neurosurgery, McGill University, Montreal, Quebec, H3G 1Y6, Canada

    Dov Frankel & Hyman M. Schipper

  6. Departments of Human Genetics, Medicine and Biology, McGill University, Montreal, Quebec, H3G 1Y6, Canada

    Leonard Pinsky

  7. Department of Medicine, McGill University, Montreal, Quebec, H3G 1Y6, Canada

    Mark A. Trifiro

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  1. Adrian Sculptoreanu
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Sculptoreanu, A., Abramovici, H., Abdullah, A.A. et al. Increased T-type Ca2+ channel activity as a determinant of cellular toxicity in neuronal cell lines expressing polyglutamine-expanded human androgen receptors. Mol Cell Biochem 203, 23–31 (2000). https://doi.org/10.1023/A:1007010020228

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