Abstract
Inappropriate apoptosis has been implicated in the mechanism of neuronal death in Huntington’s disease (HD). In this study, we report the expression of apoptotic markers in HD caudate nucleus (grades 1–4) and compare this with controls without neurological disease. Terminal transferase-mediated biotinylated-UTP nick end-labeling (TUNEL)-positive cells were detected in both control and HD brains. However, typical apoptotic cells were present only in HD, especially in grade 3 and 4 specimens. Expression of the pro-apoptotic protein Bax was increased in HD brains compared to controls, demonstrating a cytoplasmic expression pattern in predominantly shrunken and dark neurons, which were most frequently seen in grades 2 and 3. Control brains displayed weak perinuclear expression of the anti-apoptotic protein Bcl-2, whereas in HD brains Bcl-2 immunoreactivity was markedly enhanced, especially in severely affected grade 4 brains, and was observed in both healthy neurons and dark neurons. Caspase-3, an executioner protease, was only found in four HD brains of different grades and was not expressed in controls. A strong neuronal and glial expression of poly(ADP-ribose) polymerase (PARP)-immunoreactivity was observed in HD brains. These data strongly suggest the involvement of apoptosis in HD. The exact apoptotic pathway occurring in HD neurodegeneration remains yet unclear. However, the presence of late apoptotic events, such as enhanced PARP expression and many TUNEL-positive cells accompanied with weak caspase-3 immunoreactivity in severely affected HD brains, suggests that caspase-mediated neuronal death only plays a minor role in HD.
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References
Ashe P, Berry M (2003) Apoptotic signaling cascades. Prog Neuropsychopharmacol Biol Psychiatry 27:199–214
Brouillet E, Conde F, Beal MF, Hantraye P (1999) Replicating Huntington’s disease phenotype in experimental animals. Prog Neurobiol 59:427–468
Charriaut-Marlangue C, Ben-Ari Y (1995) A cautionary note on the use of the TUNEL stain to determine apoptosis. Neuroreport 7:61–64
Chen M, Ona V, Li M, Ferrante R, Fink K, Zhu S, Bian J, Guo L, Farrell L, Hersch S, Hobbs W, Vonsattel J, Cha J, Friedlander R (2000) Minocycline inhibits caspase-1 and caspase-3 expression and delays mortality in a transgenic mouse model of Huntington disease. Nat Med 6:797–801
Cookson M, Ince P, Usher P, Shaw P (1999) Poly(ADP-ribose) polymerase is found in both the nucleus and cytoplasm of human CNS neurons. Brain Res 834:182–185
Cotman, C, Anderson, A (1995) A potential role for apoptosis in neurodegeneration and Alzheimer’s disease. Mol Neurobiol 10:19–45
Dragunow M, Faull RL, Lawlor P, Beilharz EJ, Singleton K, Walker EB, Mee E (1995) In situ evidence for DNA fragmentation in Huntington’s disease striatum and Alzheimer’s disease temporal lobes. Neuroreport 6:1053–1057
Ferrer I, Planas A (2003) Signaling of cell death and cell survival following focal cerebral ischemia: life and death struggle in the penumbra. J Neuropathol Exp Neurol 62:329–339
Ferrer I, Lopez E, Blanco R, Rivera R, Ballabriga J, Pozas E, Marti E (1998) Bcl-2, Bax, and Bcl-x expression in the CA1 area of the hippocampus following transient forebrain ischemia in the adult gerbil. Exp Brain Res 121:167–173
Goldberg Y, Nicholson D, Rasper D, Kalchman M, Koide H, Graham R, Bromm M, Kazemi-Esfarjani P, Thornberry N, Vaillancourt J, Hayden M (1996) Cleavage of huntingtin by apopain, a proapoptotic cysteine protease, is modulated by the polyglutamine tract. Nat Genet 13:442–449
Grasl-Kraupp B, Ruttkay-Nedecky B, Koudelka H, Bukowska K, Bursch W, Schulte-Hermann R (1995) In situ detection of fragmented DNA (TUNEL assay) fails to discriminate among apoptosis, necrosis, and autolytic cell death: a cautionary note. Hepatology 21:1465–1468
Hara A, Hirose Y, Wang A, Yoshimi N, Tanaka T, Mori H (1996) Localization of Bax and Bcl-2 proteins, regulators of programmed cell death, in the human central nervous system. Virchows Arch 429:249–253
Hickey M, Chesselet M (2003) Apoptosis in Huntington’s disease. Prog Neuropsychopharmacol Biol Psychiatry 27:255–265
Huntington’s Disease Collaborative Research Group (1993) A novel gene containing a trinucleotide repeat that is expanded and unstable on Huntington’s disease chromosomes. Cell 72:971–983
Huppertz B, Frank H, Kaufmann P (1999) The apoptosis cascade-morphological and immunohistochemical methods for its visualization. Anat Embryol (Berl) 200:1–18
Isenmann S, Stoll G, Schroeter M, Krajewski S, Reed J, Bahr M (1998) Differential regulation of Bax, Bcl-2, and Bcl-X proteins in focal cortical ischemia in the rat. Brain Pathol 8:49–62
Kaneda K, Kashii S, Kurosawa T, Kaneko S, Akaike A, Honda Y, Minami M, Satoh M (1999) Apoptotic DNA fragmentation and upregulation of Bax induced by transient ischemia of the rat retina. Brain Res 815:11–20
Kerr JF, Wyllie AH, Curie AR (1972) Apoptosis: A basic biological phenomenon with wide-ranging implications in tissue kinetics. Br J Cancer 26:239–257
Kiechle T, Dedeoglu A, Kubilus J, Kowall NW, Beal MF, Friedlander R, Hersch SM, Ferrante RJ (2002) Cytochrome C and caspase-9 expression in Huntington’s disease. Neuromolecular Med 1:183–195
Kim S, Henkel J, Beers D, Sengun I, Simpson E, Goodman J, Engelhardt J, Siklos L, Appel S (2003) PARP expression is increased in astrocytes but decreased in motor neurons in the spinal cord of sporadic ALS patients. J Neuropathol Exp Neurol 62:88–103
Krajewski S, Krajewska M, Shabaik A, Miyashita T, Wang H, Reed J (1994) Immunohistochemical determination of in vivo distribution of Bax, a dominant inhibitor of Bcl-2. Am J Pathol 145:1323–1336
Li M, Ona VO, Chen M, Kaul M, Tenneti L, Zhang X, Stieg PE, Lipton SA, Friedlander RM (2000) Functional role and therapeutic implications of neuronal caspase-1 and - 3 in a mouse model of traumatic spinal cord injury. Neuroscience 99:333–342
Love S (2003) Apoptosis and brain ischaemia. Prog Neuropsychopharmacol Biol Psychiatry 27:267–282
Love S, Barber R, Wilcock G (1999) Increased poly(ADP-ribosyl)ation of nuclear proteins in Alzheimer’s disease. Brain 122:247–253
MacGibbon G, Lawlor P, Sirimanne E, Walton M, Connor B, Young D, Williams C, Gluckman P, Faull R, Hughes P, Dragunow M (1997) Bax expression in mammalian neurons undergoing apoptosis, and in Alzheimer’s disease hippocampus. Brain Res 750:223–234
Mandir A, Przedborski S, Jackson-Lewis V, Wang Z, Simbulan-Rosenthal C, Smulson M, Hoffman B, Guastella D, Dawson V, Dawson T (1999) Poly(ADP-ribose) polymerase activation mediates 1-methyl-4-phenyl-1, 2,3,6-tetrahydropyridine (MPTP)-induced parkinsonism. Proc Natl Acad Sci USA 96:5774–5779
Merry D, Korsmeyer S (1997) Bcl-2 gene family in the nervous system. Annu Rev Neurosci 20:245–267
Mogi M, Harada M, Kondo T, Mizuno Y, Narabayashi H, Riederer P, Nagatsu T (1996) Bcl-2 protein is increased in the brain from parkinsonian patients. Neurosci Lett 215:137–139
Nakasu S, Nakajima M, Nakazawa T, Nakasu Y, Handa J (1998) Alteration of bcl-2 and bax expression in embolized meningiomas. Brain Tumor Pathol 15:13–17
Ona V, Li M, Vonsattel J, Andrews L, Khan S, Chung W, Frey A, Menon A, Li X, Stieg P, Yuan J, Penney J, Young A, Cha J, Friedlander R (1999) Inhibition of caspase-1 slows disease progression in a mouse model of Huntington’s disease. Nature 399:263–267
Pasinelli P, Houseweart M, Brown RJ, Cleveland D (2000) Caspase-1 and −3 are sequentially activated in motor neuron death in Cu,Zn superoxide dismutase-mediated familial amyotrophic lateral sclerosis. Proc Natl Acad Sci USA 97:13901–13096
Petersen A, Mani K, Brundin P (1999) Recent advances on the pathogenesis of Huntington’s disease. Exp Neurol 157:1–18
Portera-Cailliau C, Hedreen JC, Price DL, Koliatsos VE (1995) Evidence for apoptotic cell death in Huntington disease and excitotoxic animal models. J Neurosci 15:3775–3787
Sanchez I, Xu CJ, Juo P, Kakizaka A, Blenis J, Yuan J (1999) Caspase-8 is required for cell death induced by expanded polyglutamine repeats. Neuron 22:623–633
Satou T, Cummings B, Cotman C (1995) Immunoreactivity for Bcl-2 protein within neurons in the Alzheimer’s disease brain increases with disease severity. Brain Res 697:35–43
Sugimoto T, Xiao C, Ichikawa H (1998) Neonatal primary neuronal death induced by capsaicin and axotomy involves an apoptotic mechanism. Brain Res 807:147–154
Tatton W, Chalmers-Redman R, Brown D, Tatton N (2003) Apoptosis in Parkinson’s disease: signals for neuronal degradation. Ann Neurol 53:S61–70
Thomas LB, Gates DJ, Richfield EK, O’Brien TF, Schweitzer JB, Steindler DA (1995) DNA end labeling (TUNEL) in Huntington’s disease and other neuropathological conditions. Exp Neurol 133:265–272
Uysal H, Cevik I, Soylemezoglu F, Elibol B, Ozdemir Y, Evrenkaya T, Saygi S, Dalkara T (2003) Is the cell death in mesial temporal sclerosis apoptotic? Epilepsia 44:778–784
Vis JC, Verbeek MM, Waal RM de, Donkelaar HJ ten, Kremer B (2001) The mitochondrial toxin 3-nitropropionic acid induces differential expression patterns of apoptosis-related markers in rat striatum. Neuropathol Appl Neurobiol 27:68–76
Vogel P, Dux E, Wiessner C (1997) Evidence of apoptosis in primary neuronal cultures after heat shock. Brain Res 764:205–213
Vonsattel JP, Myers RH, Stevens TJ, Ferrante RJ, Bird ED, Richardson EP, Jr (1985) Neuropathological classification of Huntington’s disease. J Neuropathol Exp Neurol 44:559–577
Acknowledgements
We thank the Department of Neurology of Leiden University Medical Center, Leiden, the Netherlands, for donation of brain tissue used in this study. This study was supported by a grant from the Netherlands Organization for Scientific Research (NWO-MW).
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Vis, J.C., Schipper, E., de Boer-van Huizen, R.T. et al. Expression pattern of apoptosis-related markers in Huntington’s disease. Acta Neuropathol 109, 321–328 (2005). https://doi.org/10.1007/s00401-004-0957-5
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DOI: https://doi.org/10.1007/s00401-004-0957-5