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Upregulation and phosphorylation of HspB1/Hsp25 and HspB5/αB-crystallin after transient middle cerebral artery occlusion in rats

  • SMALL HEAT SHOCK PROTEINS
  • Published:
Cell Stress and Chaperones Aims and scope

Abstract

Ischemic stroke leads to cellular dysfunction, cell death, and devastating clinical outcomes. The cells of the brain react to such a cellular stress by a stress response with an upregulation of heat shock proteins resulting in activation of endogenous neuroprotective capacities. Several members of the family of small heat shock proteins (HspBs) have been shown to be neuroprotective. However, yet no systematic study examined all HspBs during cerebral ischemia. Here, we performed a comprehensive comparative study comprising all HspBs in an animal model of stroke, i.e., 1 h transient middle cerebral artery occlusion followed by 23 h of reperfusion. On the mRNA level out of the 11 HspBs investigated, HspB1/Hsp25, HspB3, HspB4/αA-crystallin, HspB5/αB-crystallin, HspB7/cvHsp, and HspB8/Hsp22 were significantly upregulated in the peri-infarct region of the cerebral cortex of infarcted hemispheres. HspB1 and HspB5 reached the highest mRNA levels and were also upregulated at the protein level, suggesting that these HspBs might be functionally most relevant. Interestingly, in the infarcted cortex, both HspB1 and HspB5 were mainly allocated to neurons and to a lesser extent to glial cells. Additionally, both proteins were found to be phosphorylated in response to ischemia. Our data suggest that among all HspBs, HspB1 and HspB5 might be most important in the neuronal stress response to ischemia/reperfusion injury in the brain and might be involved in neuroprotection.

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References

  • Akbar MT, Lundberg AM, Liu K, Vidyadaran S, Wells KE, Dolatshad H, Wynn S, Wells DJ, Latchman DS, de Belleroche J (2003) The neuroprotective effects of heat shock protein 27 overexpression in transgenic animals against kainate-induced seizures and hippocampal cell death. J Biol Chem 278:19956–19965

    Article  CAS  PubMed  Google Scholar 

  • Arac A, Brownell SE, Rothbard JB, Chen C, Ko RM, Pereira MP, Albers GW, Steinman L, Steinberg GK (2011) Systemic augmentation of alphaB-crystallin provides therapeutic benefit twelve hours post-stroke onset via immune modulation. Proc Natl Acad Sci U S A 108:13287–13292

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Badin RA, Lythgoe MF, van der Weerd L, Thomas DL, Gadian DG, Latchman DS (2006) Neuroprotective effects of virally delivered HSPs in experimental stroke. J Cereb Blood Flow Metab 26:371–381

    Article  CAS  PubMed  Google Scholar 

  • Bartelt-Kirbach B, Golenhofen N (2014) Reaction of small heat-shock proteins to different kinds of cellular stress in cultured rat hippocampal neurons. Cell Stress Chaperones 19:145–153

    Article  CAS  PubMed  Google Scholar 

  • Bartelt-Kirbach B, Moron M, Glomb M, Beck CM, Weller MP, Golenhofen N (2016) HspB5/alphaB-crystallin increases dendritic complexity and protects the dendritic arbor during heat shock in cultured rat hippocampal neurons. Cell Mol Life Sci 73:3761–3775

  • Bellyei S, Szigeti A, Pozsgai E, Boronkai A, Gomori E, Hocsak E, Farkas R, Sumegi B, Gallyas F Jr (2007) Preventing apoptotic cell death by a novel small heat shock protein. Eur J Cell Biol 86:161–171

    Article  CAS  PubMed  Google Scholar 

  • Bhat SP, Nagineni CN (1989) Alpha B subunit of lens-specific protein alpha-crystallin is present in other ocular and non-ocular tissues. Biochem Biophys Res Commun 158:319–325

    Article  CAS  PubMed  Google Scholar 

  • Dang J, Mitkari B, Kipp M, Beyer C (2011) Gonadal steroids prevent cell damage and stimulate behavioral recovery after transient middle cerebral artery occlusion in male and female rats. Brain Behav Immun 25:715–726

    Article  CAS  PubMed  Google Scholar 

  • Golenhofen N, Bartelt-Kirbach B (2016) The impact of small heat shock proteins (HspBs) in Alzheimer’s and other neurological diseases. Curr Pharm Des 22:4050–4062

  • Golenhofen N, Ness W, Koob R, Htun P, Schaper W, Drenckhahn D (1998) Ischemia-induced phosphorylation and translocation of stress protein alpha B-crystallin to Z lines of myocardium. Am J Phys 274:H1457–H1464

    CAS  Google Scholar 

  • Golenhofen N, Htun P, Ness W, Koob R, Schaper W, Drenckhahn D (1999) Binding of the stress protein alpha B-crystallin to cardiac myofibrils correlates with the degree of myocardial damage during ischemia/reperfusion in vivo. J Mol Cell Cardiol 31:569–580

    Article  CAS  PubMed  Google Scholar 

  • Golenhofen N, Redel A, Wawrousek EF, Drenckhahn D (2006) Ischemia-induced increase of stiffness of alphaB-crystallin/HSPB2-deficient myocardium. Pflugers Arch 451:518–525

    Article  CAS  PubMed  Google Scholar 

  • Gong Y, Hua Y, Keep RF, Hoff JT, Xi G (2004) Intracerebral hemorrhage: effects of aging on brain edema and neurological deficits. Stroke 35:2571–2575

    Article  PubMed  Google Scholar 

  • Haslbeck M, Franzmann T, Weinfurtner D, Buchner J (2005) Some like it hot: the structure and function of small heat-shock proteins. Nat Struct Mol Biol 12:842–846

    Article  CAS  PubMed  Google Scholar 

  • Heinzel W, Vogt A, Kallee E, Faller W (1965) A new method for the quantitative determination of antibody and antigen protein, with a sensitivity to five micrograms. J Lab Clin Med 66:334–340

    CAS  PubMed  Google Scholar 

  • Horwitz J (2000) The function of alpha-crystallin in vision. Semin Cell Dev Biol 11:53–60

    Article  CAS  PubMed  Google Scholar 

  • Imura T, Shimohama S, Sato M, Nishikawa H, Madono K, Akaike A, Kimura J (1999) Differential expression of small heat shock proteins in reactive astrocytes after focal ischemia: possible role of beta-adrenergic receptor. J Neurosci 19:9768–9779

    CAS  PubMed  Google Scholar 

  • Kampinga HH, Hageman J, Vos MJ, Kubota H, Tanguay RM, Bruford EA, Cheetham ME, Chen B, Hightower LE (2009) Guidelines for the nomenclature of the human heat shock proteins. Cell Stress Chaperones 14:105–111

    Article  CAS  PubMed  Google Scholar 

  • Kamradt MC, Chen F, Sam S, Cryns VL (2002) The small heat shock protein alpha B-crystallin negatively regulates apoptosis during myogenic differentiation by inhibiting caspase-3 activation. J Biol Chem 277:38731–38736

    Article  CAS  PubMed  Google Scholar 

  • Kappe G, Franck E, Verschuure P, Boelens WC, Leunissen JA, de Jong WW (2003) The human genome encodes 10 alpha-crystallin-related small heat shock proteins: HspB1-10. Cell Stress Chaperones 8:53–61

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Kappe G, Boelens WC, de Jong WW (2010) Why proteins without an alpha-crystallin domain should not be included in the human small heat shock protein family HSPB. Cell Stress Chaperones 15:457–461

    Article  CAS  PubMed  Google Scholar 

  • Kato H, Liu Y, Kogure K, Kato K (1994) Induction of 27-kDa heat shock protein following cerebral ischemia in a rat model of ischemic tolerance. Brain Res 634:235–244

    Article  CAS  PubMed  Google Scholar 

  • Kato K, Ito H, Kamei K, Inaguma Y, Iwamoto I, Saga S (1998) Phosphorylation of alphaB-crystallin in mitotic cells and identification of enzymatic activities responsible for phosphorylation. J Biol Chem 273:28346–28354

    Article  CAS  PubMed  Google Scholar 

  • Kato K, Katoh-Semba R, Takeuchi IK, Ito H, Kamei K (1999) Responses of heat shock proteins hsp27, alphaB-crystallin, and hsp70 in rat brain after kainic acid-induced seizure activity. J Neurochem 73:229–236

    Article  CAS  PubMed  Google Scholar 

  • Kirbach BB, Golenhofen N (2011) Differential expression and induction of small heat shock proteins in rat brain and cultured hippocampal neurons. J Neurosci Res 89:162–175

    Article  CAS  PubMed  Google Scholar 

  • Kostenko S, Moens U (2009) Heat shock protein 27 phosphorylation: kinases, phosphatases, functions and pathology. Cell Mol Life Sci 66:3289–3307

    Article  CAS  PubMed  Google Scholar 

  • MacRae TH (2000) Structure and function of small heat shock/alpha-crystallin proteins: established concepts and emerging ideas. Cell Mol Life Sci 57:899–913

    Article  CAS  PubMed  Google Scholar 

  • Mehlen P, Schulze-Osthoff K, Arrigo AP (1996) Small stress proteins as novel regulators of apoptosis. Heat shock protein 27 blocks Fas/APO-1- and staurosporine-induced cell death. J Biol Chem 271:16510–16514

    Article  CAS  PubMed  Google Scholar 

  • Oshita SE, Chen F, Kwan T, Yehiely F, Cryns VL (2010) The small heat shock protein HspB2 is a novel anti-apoptotic protein that inhibits apical caspase activation in the extrinsic apoptotic pathway. Breast Cancer Res Treat 124:307–315

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Ousman SS, Tomooka BH, van Noort JM, Wawrousek EF, O’Connor KC, Hafler DA, Sobel RA, Robinson WH, Steinman L (2007) Protective and therapeutic role for alphaB-crystallin in autoimmune demyelination. Nature 448:474–479

    Article  CAS  PubMed  Google Scholar 

  • Piao CS, Kim SW, Kim JB, Lee JK (2005) Co-induction of αB-crystallin and MAPKAPK-2 in astrocytes in the penumbra after transient focal cerebral ischemia. Exp Brain Res 163(4):421–429

  • Popp A, Jaenisch N, Witte OW, Frahm C (2009) Identification of ischemic regions in a rat model of stroke. PLoS One 4:e4764

    Article  PubMed  PubMed Central  Google Scholar 

  • Quraishe S, Asuni A, Boelens WC, O’Connor V, Wyttenbach A (2008) Expression of the small heat shock protein family in the mouse CNS: Differential anatomical and biochemical compartmentalization. Neuroscience 153:483–491

    Article  CAS  PubMed  Google Scholar 

  • Ray PS, Martin JL, Swanson EA, Otani H, Dillmann WH, Das DK (2001) Transgene overexpression of αB crystallin confers simultaneous protection against cardiomyocyte apoptosis and necrosis during myocardial ischemia and reperfusion. FASEB J 15:393–402

    Article  CAS  PubMed  Google Scholar 

  • Ritossa FM (1962) A new puffing pattern induced by a temperature shock and DNP in Drosophila. Experientia 18:571–573

    Article  CAS  Google Scholar 

  • Schmidt T, Bartelt-Kirbach B, Golenhofen N (2012) Phosphorylation-dependent subcellular localization of the small heat shock proteins HspB1/Hsp25 and HspB5/alphaB-crystallin in cultured hippocampal neurons. Histochem Cell Biol 138:407–418

    Article  CAS  PubMed  Google Scholar 

  • Shimada Y, Tanaka R, Shimura H, Yamashiro K, Urabe T, Hattori N (2014) Phosphorylation enhances recombinant HSP27 neuroprotection against focal cerebral ischemia in mice. Neuroscience 278:113–121

    Article  CAS  PubMed  Google Scholar 

  • Stetler RA, Cao G, Gao Y et al (2008) Hsp27 protects against ischemic brain injury via attenuation of a novel stress-response cascade upstream of mitochondrial cell death signaling. J Neurosci 28:13038–13055

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Stetler RA, Gan Y, Zhang W, Liou AK, Gao Y, Cao G, Chen J (2010) Heat shock proteins: cellular and molecular mechanisms in the central nervous system. Prog Neurobiol 92:184–211

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Stetler RA, Gao Y, Zhang L et al (2012) Phosphorylation of HSP27 by protein kinase D is essential for mediating neuroprotection against ischemic neuronal injury. J Neurosci 32:2667–2682

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Sugiyama Y, Suzuki A, Kishikawa M, Akutsu R, Hirose T, Waye MM, Tsui SK, Yoshida S, Ohno S (2000) Muscle develops a specific form of small heat shock protein complex composed of MKBP/HSPB2 and HSPB3 during myogenic differentiation. J Biol Chem 275:1095–1104

    Article  CAS  PubMed  Google Scholar 

  • Tao X, Lu W, Deng J, Hu Z, Lei Q, Zhang J, Song T, Liu J, Zheng L, He J (2015) HspB8 expression in brain tissue after cerebral ischemic reperfusion and atorvastatin intervention in Sprague-Dawley rats. Neurol Res 37:229–237

    Article  CAS  PubMed  Google Scholar 

  • Teramoto S, Shimura H, Tanaka R, Shimada Y, Miyamoto N, Arai H, Urabe T, Hattori N (2013) Human-derived physiological heat shock protein 27 complex protects brain after focal cerebral ischemia in mice. PLoS One 8:e66001

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Ulbrich C, Zendedel A, Habib P, Kipp M, Beyer C, Dang J (2012) Long-term cerebral cortex protection and behavioral stabilization by gonadal steroid hormones after transient focal hypoxia. J Steroid Biochem Mol Biol 131:10–16

    Article  CAS  PubMed  Google Scholar 

  • van der Weerd L, Tariq Akbar M, Aron Badin R, Valentim LM, Thomas DL, Wells DJ, Latchman DS, Gadian DG, Lythgoe MF, de Belleroche JS (2010) Overexpression of heat shock protein 27 reduces cortical damage after cerebral ischemia. J Cereb Blood Flow Metab 30:849–856

    Article  PubMed  Google Scholar 

  • Wagstaff MJ, Collaco-Moraes Y, Smith J, de Belleroche JS, Coffin RS, Latchman DS (1999) Protection of neuronal cells from apoptosis by Hsp27 delivered with a herpes simplex virus-based vector. J Biol Chem 274:5061–5069

    Article  CAS  PubMed  Google Scholar 

  • Welch WJ (1992) Mammalian stress response: cell physiology, structure/function of stress proteins, and implications for medicine and disease. Physiol Rev 72:1063–1081

    CAS  PubMed  Google Scholar 

  • Yang B, Zhang H, Mo X, Xiao H, Hu Z (2015) HspB8 is neuroprotective during oxygen glucose deprivation and reperfusion. Curr Neurovasc Res 12:63–72

    Article  CAS  PubMed  Google Scholar 

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Acknowledgements

We thank Bianca Mekle, Diana Reinhard, and Stephanie Sues for their excellent technical assistance.

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Authors and Affiliations

  1. Institute of Anatomy and Cell Biology, University of Ulm, Albert-Einstein-Allee 11, 89081, Ulm, Germany

    Britta Bartelt-Kirbach & Nikola Golenhofen

  2. Institute of Neuroanatomy, RWTH Aachen University, Aachen, Germany

    Alexander Slowik & Cordian Beyer

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  1. Britta Bartelt-Kirbach
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  2. Alexander Slowik
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  4. Nikola Golenhofen
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Correspondence to Nikola Golenhofen.

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Animal care and experimental procedures were formally approved by the Review Board for the Care of Animal Subjects of the district government (North Rhine-Westphalia, Germany) and bred and maintained in a pathogen-free environment in a 12-h day and 12-h night cycle prior experiments.

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Bartelt-Kirbach, B., Slowik, A., Beyer, C. et al. Upregulation and phosphorylation of HspB1/Hsp25 and HspB5/αB-crystallin after transient middle cerebral artery occlusion in rats. Cell Stress and Chaperones 22, 653–663 (2017). https://doi.org/10.1007/s12192-017-0794-9

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  • DOI: https://doi.org/10.1007/s12192-017-0794-9

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