Abstract
Clusterin, a protein chaperone found at high levels in physiological fluids, is expressed in nervous tissue and upregulated in several neurological diseases. To assess relevance to amyotrophic lateral sclerosis (ALS) and other motor neuron disorders, clusterin expression was evaluated using long-term dissociated cultures of murine spinal cord and SOD1G93A transgenic mice, a model of familial ALS. Motor neurons and astrocytes constitutively expressed nuclear and cytoplasmic forms of clusterin, and secreted clusterin accumulated in culture media. Although clusterin can be stress inducible, heat shock failed to increase levels in these neural cell compartments despite robust upregulation of stress-inducible Hsp70 (HspA1) in non-neuronal cells. In common with HSPs, clusterin was upregulated by treatment with the Hsp90 inhibitor, geldanamycin, and thus could contribute to the neuroprotection previously identified for such compounds in disease models. Clusterin expression was not altered in cultured motor neurons expressing SOD1G93A by gene transfer or in presymptomatic SOD1G93A transgenic mice; however, clusterin immunolabeling was weakly increased in lumbar spinal cord of overtly symptomatic mice. More striking, mutant SOD1 inclusions, a pathological hallmark, were strongly labeled by anti-clusterin. Since secreted, as well as intracellular, mutant SOD1 contributes to toxicity, the extracellular chaperoning property of clusterin could be important for folding and clearance of SOD1 and other misfolded proteins in the extracellular space. Evaluation of chaperone-based therapies should include evaluation of clusterin as well as HSPs, using experimental models that replicate the control mechanisms operant in the cells and tissue of interest.
Similar content being viewed by others
References
Batulan Z, Shinder GA, Minotti S, He BP, Doroudchi MM, Nalbantoglu J, Strong MJ, Durham HD (2003) High threshold for induction of the stress response in motor neurons is associated with failure to activate HSF1. J Neurosci 23:5789–5798
Batulan Z, Taylor DM, Aarons RJ, Minotti S, Doroudchi MM, Nalbantoglu J, Durham HD (2006) Induction of multiple heat shock proteins and neuroprotection in a primary culture model of familial amyotrophic lateral sclerosis. Neurobiol Dis 24:213–225
Boillee S, Vande VC, Cleveland DW (2006) ALS: a disease of motor neurons and their nonneuronal neighbors. Neuron 52:39–59
Bonnard AS, Chan P, Fontaine M (1997) Expression of clusterin and C4 mRNA during rat peripheral nerve regeneration. Immunopharmacology 38:81–86
Broquet AH, Thomas G, Masliah J, Trugnan G, Bachelet M (2003) Expression of the molecular chaperone Hsp70 in detergent-resistant microdomains correlates with its membrane delivery and release. J Biol Chem 278:21601–21606
Caccamo AE, Desenzani S, Belloni L, Borghetti AF, Bettuzzi S (2006) Nuclear clusterin accumulation during heat shock response: implications for cell survival and thermo-tolerance induction in immortalized and prostate cancer cells. J Cell Physiol 207:208–219
Charnay Y, Imhof A, Vallet PG, Hakkoum D, Lathuiliere A, Poku N, Aronow B, Kovari E, Bouras C, Giannakopoulos P (2008) Clusterin expression during fetal and postnatal CNS development in mouse. Neuroscience 155:714–724
Clark AM, Griswold MD (1997) Expression of clusterin/sulfated glycoprotein-2 under conditions of heat stress in rat Sertoli cells and a mouse Sertoli cell line. J Androl 18:257–263
Danik M, Chabot JG, Hassan-Gonzalez D, Suh M, Quirion R (1993) Localization of sulfated glycoprotein-2/clusterin mRNA in the rat brain by in situ hybridization. J Comp Neurol 334:209–227
Debure L, Vayssiere JL, Rincheval V, Loison F, Le DY, Michel D (2003) Intracellular clusterin causes juxtanuclear aggregate formation and mitochondrial alteration. J Cell Sci 116:3109–3121
Didelot C, Lanneau D, Brunet M, Bouchot A, Cartier J, Jacquel A, Ducoroy P, Cathelin S, Decologne N, Chiosis G, Dubrez-Daloz L, Solary E, Garrido C (2008) Interaction of heat-shock protein 90 beta isoform (HSP90 beta) with cellular inhibitor of apoptosis 1 (c-IAP1) is required for cell differentiation. Cell Death Differ 15:859–866
Dragunow M, Preston K, Dodd J, Young D, Lawlor P, Christie D (1995) Clusterin accumulates in dying neurons following status epilepticus. Brain Res Mol Brain Res 32:279–290
Durham HD, Roy J, Dong L, Figlewicz DA (1997) Aggregation of mutant Cu/Zn superoxide dismutase proteins in a culture model of ALS. J Neuropathol Exp Neurol 56:523–530
Evdokimovskaya Y, Skarga Y, Vrublevskaya V, Morenkov O (2010) Secretion of the heat shock proteins HSP70 and HSC70 by baby hamster kidney (BHK-21) cells. Cell Biol Int 34:985–990
Falgarone G, Chiocchia G (2009) Chapter 8: Clusterin: a multifacet protein at the crossroad of inflammation and autoimmunity. Adv Cancer Res 104:139–170
Ghiso J, Matsubara E, Koudinov A, Choi-Miura NH, Tomita M, Wisniewski T, Frangione B (1993) The cerebrospinal-fluid soluble form of Alzheimer's amyloid beta is complexed to SP-40,40 (apolipoprotein J), an inhibitor of the complement membrane-attack complex. Biochem J 293(Pt 1):27–30
Giannakopoulos P, Kovari E, French LE, Viard I, Hof PR, Bouras C (1998) Possible neuroprotective role of clusterin in Alzheimer's disease: a quantitative immunocytochemical study. Acta Neuropathol 95:387–394
Grewal RP, Morgan TE, Finch CE (1999) C1qB and clusterin mRNA increase in association with neurodegeneration in sporadic amyotrophic lateral sclerosis. Neurosci Lett 271:65–67
Gurney ME (1994) Transgenic-mouse model of amyotrophic lateral sclerosis. N Engl J Med 331:1721–1722
Gurney ME, Pu H, Chiu AY, Dal Canto MC, Polchow CY, Alexander DD, Caliendo J, Hentati A, Kwon YW, Deng H-X, Chen W, Zhai P, Sufit RL, Siddique T (1994) Motor neuron degeneration in mice that express a human Cu, Zn superoxide dismutase mutation. Science 264:1772–1775
Guzhova I, Kislyakova K, Moskaliova O, Fridlanskaya I, Tytell M, Cheetham M, Margulis B (2001) In vitro studies show that Hsp70 can be released by glia and that exogenous Hsp70 can enhance neuronal stress tolerance. Brain Res 914:66–73
Hightower LE, Guidon PT Jr (1989) Selective release from cultured mammalian cells of heat-shock (stress) proteins that resemble glia-axon transfer proteins. J Cell Physiol 138:257–266
Kalmar B, Burnstock G, Vrbova G, Greensmith L (2002) The effect of neonatal nerve injury on the expression of heat shock proteins in developing rat motoneurones. J Neurotrauma 19:667–679
Leskov KS, Klokov DY, Li J, Kinsella TJ, Boothman DA (2003) Synthesis and functional analyses of nuclear clusterin, a cell death protein. J Biol Chem 278:11590–11600
Lidstrom AM, Bogdanovic N, Hesse C, Volkman I, Davidsson P, Blennow K (1998) Clusterin (apolipoprotein J) protein levels are increased in hippocampus and in frontal cortex in Alzheimer's disease. Exp Neurol 154:511–521
Loison F, Debure L, Nizard P, Le GP, Michel D, Le DY (2006) Up-regulation of the clusterin gene after proteotoxic stress: implication of HSF1-HSF2 heterocomplexes. Biochem J 395:223–231
Mambula SS, Calderwood SK (2006) Heat shock protein 70 is secreted from tumor cells by a nonclassical pathway involving lysosomal endosomes. J Immunol 177:7849–7857
Manzerra P, Brown IR (1992) Expression of heat shock genes (hsp70) in the rabbit spinal cord: Localization of constitutive and hyperthermia-inducible mRNA species. J Neurosci Res 31:606–615
Manzerra P, Brown IR (1996) The neuronal stress response: nuclear translocation of heat shock proteins as an indicator of hyperthermic stress. Exp Cell Res 229:35–47
May PC, Johnson SA, Poirier J, Lampert-Etchells M, Finch CE (1989) Altered gene expression in Alzheimer's disease brain tissue. Can J Neurol Sci 16:473–476
May PC, Robison P, Fuson K, Smalstig B, Stephenson D, Clemens JA (1992) Sulfated glycoprotein-2 expression increases in rodent brain after transient global ischemia. Brain Res Mol Brain Res 15:33–39
Murakami M, Ushio Y, Morino Y, Ohta T, Matsukado Y (1988) Immunohistochemical localization of apolipoprotein E in human glial neoplasms. J Clin Invest 82:177–188
Nizard P, Tetley S, Le DY, Watrin T, Le GP, Wilson MR, Michel D (2007) Stress-induced retrotranslocation of clusterin/ApoJ into the cytosol. Traffic 8:554–565
Ohlsson M, Bellander BM, Langmoen IA, Svensson M (2003) Complement activation following optic nerve crush in the adult rat. J Neurotrauma 20:895–904
Pasinetti GM, Johnson SA, Oda T, Rozovsky I, Finch CE (1994) Clusterin (SGP-2): a multifunctional glycoprotein with regional expression in astrocytes and neurons of the adult rat brain. J Comp Neurol 339:387–400
Polihronis M, Paizis K, Carter G, Sedal L, Murphy B (1993) Elevation of human cerebrospinal fluid clusterin concentration is associated with acute neuropathology. J Neurol Sci 115:230–233
Poon S, Easterbrook-Smith SB, Rybchyn MS, Carver JA, Wilson MR (2000) Clusterin is an ATP-independent chaperone with very broad substrate specificity that stabilizes stressed proteins in a folding-competent state. Biochemistry 39:15953–15960
Rithidech KN, Honikel L, Milazzo M, Madigan D, Troxell R, Krupp LB (2009) Protein expression profiles in pediatric multiple sclerosis: potential biomarkers. Mult Scler 15:455–464
Rizzi F, Coletta M, Bettuzzi S (2009) Chapter 2: Clusterin (CLU): from one gene and two transcripts to many proteins. Adv Cancer Res 104:9–23
Robinson MB, Tidwell JL, Gould T, Taylor AR, Newbern JM, Graves J, Tytell M, Milligan CE (2005) Extracellular heat shock protein 70: a critical component for motoneuron survival. J Neurosci 25:9735–9745
Rosen DR, Siddique T, Patterson D, Figlewicz DA, Sapp P, Hentati A, Donaldson D, Goto J, O'Regan JP, Deng H-X, Rahmani Z, Krizus A, McKenna-Yasek D, Cayabyab A, Gaston SM, Berger R, Tanzi RE, Halperin JJ, Herzfeldt B, Van den Bergh R, Hung W-Y, Bird T, Deng G, Mulder DW, Smyth C, Laing NG, Soriano E, Pericak-Vance MA, Haines J, Rouleau GA, Gusella JS, Horvitz HR, Brown RH Jr (1993) Mutations in Cu/Zn superoxide dismutase gene are associated with familial amyotrophic lateral sclerosis. Nature 362:59–62
Roy J, Minotti S, Dong L, Figlewicz DA, Durham HD (1998) Glutamate potentiates the toxicity of mutant Cu/Zn-superoxide dismutase in motor neurons by postsynaptic calcium-dependent mechanisms. J Neurosci 18:9673–9684
Schreiber SS, Tocco G, Najm I, Baudry M (1993) Seizure activity causes a rapid increase in sulfated glycoprotein-2 messenger RNA in the adult but not the neonatal rat brain. Neurosci Lett 153:17–20
Taylor DM, De KP, Minotti S, Durham HD (2007a) Manipulation of protein kinases reveals different mechanisms for upregulation of heat shock proteins in motor neurons and non-neuronal cells. Mol Cell Neurosci 34:20–33
Taylor DM, Tradewell ML, Minotti S, Durham HD (2007b) Characterizing the role of Hsp90 in production of heat shock proteins in motor neurons reveals a suppressive effect of wild-type Hsf1. Cell Stress Chaperones 12:151–162
Trougakos IP, Gonos ES (2006) Regulation of clusterin/apolipoprotein J, a functional homologue to the small heat shock proteins, by oxidative stress in ageing and age-related diseases. Free Radic Res 40:1324–1334
Turner BJ, Atkin JD, Farg MA, Zang dW, Rembach A, Lopes EC, Patch JD, Hill AF, Cheema SS (2005) Impaired extracellular secretion of mutant superoxide dismutase 1 associates with neurotoxicity in familial amyotrophic lateral sclerosis. J Neurosci 25:108–117
Urushitani M, Sik A, Sakurai T, Nukina N, Takahashi R, Julien JP (2006) Chromogranin-mediated secretion of mutant superoxide dismutase proteins linked to amyotrophic lateral sclerosis. Nat Neurosci 9:108–118
Van Beek J, Chan P, Bernaudin M, Petit E, MacKenzie ET, Fontaine M (2000) Glial responses, clusterin, and complement in permanent focal cerebral ischemia in the mouse. Glia 31:39–50
Villoslada P, Moreno B, Melero I, Pablos JL, Martino G, Uccelli A, Montalban X, Avila J, Rivest S, Acarin L, Appel S, Khoury SJ, McGeer P, Ferrer I, Delgado M, Obeso J, Schwartz M (2008) Immunotherapy for neurological diseases. Clin Immunol 128:294–305
Voellmy R (2004) On mechanisms that control heat shock transcription factor activity in metazoan cells. Cell Stress Chaperones 9:122–133
Watanabe M, Dykes-Hoberg M, Culotta VC, Price DL, Wong PC, Rothstein JD (2001) Histological evidence of protein aggregation in mutant SOD1 transgenic mice and in amyotrophic lateral sclerosis neural tissues. Neurobiol Dis 8:933–941
Wiggins AK, Shen PJ, Gundlach AL (2003) Delayed, but prolonged increases in astrocytic clusterin (ApoJ) mRNA expression following acute cortical spreading depression in the rat: evidence for a role of clusterin in ischemic tolerance. Brain Res Mol Brain Res 114:20–30
Wilson MR, Easterbrook-Smith SB (2000) Clusterin is a secreted mammalian chaperone. Trends Biochem Sci 25:95–98
Yang CR, Leskov K, Hosley-Eberlein K, Criswell T, Pink JJ, Kinsella TJ, Boothman DA (2000) Nuclear clusterin/XIP8, an x-ray-induced Ku70-binding protein that signals cell death. Proc Natl Acad Sci U S A 97:5907–5912
Zhang H, Kim JK, Edwards CA, Xu Z, Taichman R, Wang CY (2005) Clusterin inhibits apoptosis by interacting with activated Bax. Nat Cell Biol 7:909–915
Acknowledgments
This work was supported by the Canadian Institutes for Health Research (MOP77743). Immunohistochemistry on mouse spinal cord sections and characterization were conducted by Cytochem, Inc., Montreal, Canada (Dr. Martin Marcinkiewicz, President and CSO) through a fee for service contract. The data obtained in culture models were included in the M.Sc. thesis of S.Z.
Author information
Authors and Affiliations
Corresponding author
Electronic supplementary material
Below is the link to the electronic supplementary material.
Online resource 1
(PDF 120 kb)
Rights and permissions
About this article
Cite this article
Zinkie, S., Gentil, B.J., Minotti, S. et al. Expression of the protein chaperone, clusterin, in spinal cord cells constitutively and following cellular stress, and upregulation by treatment with Hsp90 inhibitor. Cell Stress and Chaperones 18, 745–758 (2013). https://doi.org/10.1007/s12192-013-0427-x
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s12192-013-0427-x