Abstract
Autophagy is a mechanism by which parts of a cell that are old and unneeded are segregated inside structures called “autophagosomes”. The materials ingested by this autophagy are brought to cellular compartments called “lysosomes,” which are specific intracellular compartments for degradation, and the degraded products are re-used for cell metabolism. We have shown that, in mice, deficiency in lysosomal proteinases such as cathepsin D or cathepsins B and L induces the accumulation of lysosomes containing ceroid-lipofuscin; the phenotypes of these mice resemble those of neuronal ceroid lipofuscinosis (NCL). In these mutant mice, the accumulation of abnormal lysosomal structures appears in accordance with an increase in the amount of membrane-bound microtubule associated protein 1 light chain 3 (LC3), a marker of “autophagosomes” in neurons. Such autophagosomes often contain granular osmiophilic deposits, a hallmark of NCL, together with part of the cytoplasm, which contains undigested materials. These data strongly argue for a major involvement of autophagy in the pathogenesis of NCL, although it remains largely unknown what signaling is essential for autophagosome formation.
Neonatal hypoxic/ischemic (H/I) brain injury causes neurological impairment, including cognitive and motor dysfunction. as well as seizures. However, the molecular mechanisms regulating neuron death after H/I injury are poorly defined and remain controversial. Here we show that Atg7, a gene essential for autophagy induction, is a critical mediator of H/I-induced neuron death. Neonatal mice subjected to H/I injury show dramatically increased autophagosome formation and extensive hippocampal neuron death that is regulated by both caspase-3-dependent and -independent execution. Mice deficient in Atg7 show nearly complete protection from both H/I-induced caspase-3 activation and neuron death, indicating that Atg7 is critically positioned upstream of multiple neuronal death executioner pathways. Adult H/I brain injury also produces a significant increase in autophagy, but, unlike neonatal H/I, neuron death is almost exclusively caspase-3-independent. These data suggest that autophagy plays an essential role in triggering neuronal death execution after H/I injury.
Although it has been considered that autophagy is essential for the maintenance of cellular metabolism, our data suggest that excess autophagy under pathological conditions may lead to cell death.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Preview
Unable to display preview. Download preview PDF.
References
Uchiyama Y, Shibata M, Koike M et al (2008) Autophagy—physiology and pathophysiology. Histochem Cell Biol 129: 407–420
Blomgren K, Leist M, Groc L (2007) Pathological apoptosis in the developing brain. Apoptosis 12: 993–1010
Blomgren K, Hagberg H (2006) Free radicals, mitochondria, and hypoxia-ischemia in the developing brain. Free Radic Biol Med 40: 388–397
Blomgren K, Zhu C, Hallin U et al (2003) Mitochondria and ischemic reperfusion damage in the adult and in the developing brain. Biochem Biophys Res Commun 304: 551–559
Koike M, Shibata M, Tadakoshi M et al (2008) Inhibition of autophagy prevents hippocampal pyramidal neuron death after hypoxic-ischemic injury. Am J Pathol 172: 454–469
Uchiyama Y, Koike M, Shibata M (2008) Autophagic neuron death in neonatal brain ischemia/hypoxia. Autophagy 4: 404–408
Uchiyama Y, Koike M, Shibata M et al (2009) Autophagic neuron death. Methods Enzymol 453: 33–51
Nitatori T, Sato N, Waguri S et al (1995) Delayed neuronal death in the CA1 pyramidal cell layer of the gerbil hippocampus following transient ischemia is apoptosis. J Neurosci 15: 1001–1011
Mizushima N (2007) Autophagy: process and function. Genes Dev 21: 2861–2873
Shintani T, Klionsky DJ (2004) Autophagy in health and disease: a double-edged sword. Science 306: 990–995
Koike M, Shibata M, Waguri S et al (2005) Participation of autophagy in storage of lysosomes in neurons from mouse models of neuronal ceroid-lipofuscinoses (Batten disease). Am J Pathol 167: 1713–1728
Nixon RA (2006) Autophagy in neurodegenerative disease: friend, foe or turncoat? Trends Neurosci 29: 528–535
Zhu C, Wang X, Xu F et al (2005) The influence of age on apoptotic and other mechanisms of cell death after cerebral hypoxia-ischemia. Cell Death Differ 12: 162–176
Kabeya Y, Mizushima N, Ueno T et al (2000) LC3, a mammalian homologue of yeast Apg8p, is localized in autophagosome membranes after processing. EMBO J 19: 5720–5728
Haltia M (2006) The neuronal ceroid-lipofuscinoses: from past to present. Biochim Biophys Acta 1762: 850–856
Ivy GO, Schottler F, Wenzel J et al (1984) Inhibitors of lysosomal enzymes: accumulation of lipofuscin-like dense bodies in the brain. Science 226: 985–987
Sleat DE, Donnelly RJ, Lackland H et al (1997) Association of mutations in a lysosomal protein with classical late-infantile neuronal ceroid lipofuscinosis. Science 277: 1802–1805
Ezaki J, Takeda-Ezaki M, Kominami E (2000) Tripeptidyl peptidase I, the late infantile neuronal ceroid lipofuscinosis gene product, initiates the lysosomal degradation of subunit c of ATP synthase. J Biochem 128: 509–516
Ezaki J, Tanida I, Kanehagi N et al (1999) A lysosomal proteinase, the late infantile neuronal ceroid lipofuscinosis gene (CLN2) product, is essential for degradation of a hydrophobic protein, the subunit c of ATP synthase. J Neurochem 72: 2573–2582
Koike M, Nakanishi H, Saftig P et al (2000) Cathepsin D deficiency induces lysosomal storage with ceroid lipofuscin in mouse CNS neurons. J Neurosci 20: 6898–6906
Koike M, Shibata M, Ohsawa Y et al (2003) Involvement of two different cell death pathways in retinal atrophy of cathepsin D-deficient mice. Mol Cell Neurosci 22: 146–161
Nakanishi H, Zhang J, Koike M et al (2001) Involvement of nitric oxide released from microglia-macrophages in pathological changes of cathepsin D-deficient mice. J Neurosci 21: 7526–7533
Siintola E, Partanen S, Stromme P et al (2006) Cathepsin D deficiency underlies congenital human neuronal ceroid-lipofuscinosis. Brain 129: 1438–1445
Steinfeld R, Reinhardt K, Schreiber K et al (2006) Cathepsin D deficiency is associated with a human neurodegenerative disorder. Am J Hum Genet 78: 988–998
Dunn WA, Jr. (1994) Autophagy and related mechanisms of lysosome-mediated protein degradation. Trends Cell Biol 4: 139–143
Dunn WA Jr (1990) Studies on the mechanisms of autophagy: maturation of the autophagic vacuole. J Cell Biol 110: 1935–1945
Liou W, Geuze HJ, Geelen MJ et al (1997) The autophagic and endocytic pathways converge at the nascent autophagic vacuoles. J Cell Biol 136: 61–70
Tanaka Y, Guhde G, Suter A et al (2000) Accumulation of autophagic vacuoles and cardiomyopathy in LAMP-2-deficient mice. Nature 406: 902–906
Hara T, Nakamura K, Matsui M et al (2006) Suppression of basal autophagy in neural cells causes neurodegenerative disease in mice. Nature 441: 885–889
Komatsu M, Waguri S, Chiba T et al (2006) Loss of autophagy in the central nervous system causes neurodegeneration in mice. Nature 441: 880–884
Komatsu M, Waguri S, Ueno T et al (2005) Impairment of starvation-induced and constitutive autophagy in Atg7-deficient mice. J Cell Biol 169: 425–434
Ardley HC, Hung CC, Robinson PA (2005) The aggravating role of the ubiquitin-proteasome system in neurodegeneration. FEBS Lett 579: 571–576
Settembre C, Fraldi A, Jahreiss L et al (2008) A block of autophagy in lysosomal storage disorders. Hum Mol Genet 17: 119–129
Zhan SS, Beyreuther K, Schmitt HP (1992) Neuronal ubiquitin and neurofilament expression in different lysosomal storage disorders. Clin Neuropathol 11: 251–255
Bjørkøy G, Lamark T, Brech A et al (2005) p62/SQSTM1 forms protein aggregates degraded by autophagy and has a protective effect on huntingtin-induced cell death. J Cell Biol 171: 603–614
Moscat J, Diaz-Meco MT, Albert A et al (2006) Cell signaling and function organized by PB1 domain interactions. Mol Cell 23: 631–640
Wooten MW, Hu X, Babu JR et al (2006) Signaling, polyubiquitination, trafficking, and inclusions: sequestosome 1/p62’s role in neurodegenerative disease. J Biomed Biotechnol 2006: 62–79
Komatsu M, Waguri S, Koike M et al (2007) Homeostatic levels of p62 control cytoplasmic inclusion body formation in autophagy-deficient mice. Cell 131: 1149–1163
Nakai A, Yamaguchi O, Takeda T et al (2007) The role of autophagy in cardiomyocytes in the basal state and in response to hemodynamic stress. Nat Med 13: 619–624
Wang QJ, Ding Y, Kohtz DS et al (2006) Induction of autophagy in axonal dystrophy and degeneration. J Neurosci 26: 8057–8068
Walker NI, Harmon BV, Gobe GC et al (1988) Patterns of cell death. Methods Achiev Exp Pathol 13: 18–54
Lockshin RA, Williams CM (1964) Programmed cell death. II. Endocrine potentiation of the breakdown of the intersegmental muscles of silkmoths. J Insect Physiol 10: 643–649
Lockshin RA, Zaleri Z (1991) Programmed cell death and apoptosis. Cold Spring Harbor Laboratory, New York
Kerr JF, Wythe AH, Currie AR (1972) Apoptosis: a basic biological phenomenon with wide-ranging implications in tissue kinetics. Br J Cancer 26: 239–257
Salvesen GS, Dixit VM (1999) Caspase activation: the induced-proximity model. Proc Natl Acad Sci U S A 96: 10 964–10 967
Enari M, Sakahira H, Yokoyama H et al (1998) A caspase-activated DNase that degrades DNA during apoptosis, and its inhibitor ICAD. Nature 391: 43–50
Li P, Nijhawan D, Budihardjo I et al (1997) Cytochrome c and dATP-dependent formation of Apaf-1/caspase-9 complex initiates an apoptotic protease cascade. Cell 91: 479–489
Nicholson DW, Thornberry NA (1997) Caspases: killer proteases. Trends Biochem Sci 22: 299–306
Sakahira H, Enari M, Nagata S (1998) Cleavage of CAD inhibitor in CAD activation and DNA degradation during apoptosis. Nature 391: 96–99
Sakahira H, Enari M, Ohsawa Y et al (1999) Apoptotic nuclear morphological change without DNA fragmentation. Curr Biol 9: 543–546
Raff MC, Barres BA, Burne JF et al (1993) Programmed cell death and the control of cell survival: lessons from the nervous system. Science 262: 695–700
Blomgren K, Zhu C, Wang X et al (2001) Synergistic activation of caspase-3 by m-calpain after neonatal hypoxia-ischemia: a mechanism of “pathological apoptosis”? J Biol Chem 276: 10 191–10 198
Gill R, Soriano M, Blomgren K et al (2002) Role of caspase-3 activation in cerebral ischemia-induced neurodegeneration in adult and neonatal brain. J Cereb Blood Flow Metab 22: 420–430
Hu BR, Liu CL, Ouyang Y et al (2000) Involvement of caspase-3 in cell death after hypoxia-ischemia declines during brain maturation. J Cereb Blood Flow Metab 20: 1294–1300
Liu CL, Siesjo BK, Hu BR (2004) Pathogenesis of hippocampal neuronal death after hypoxia-ischemia changes during brain development. Neuroscience 127: 113–123
Zhu C, Qiu L, Wang X et al (2003) Involvement of apoptosis-inducing factor in neuronal death after hypoxia-ischemia in the neonatal rat brain. J Neurochem 86: 306–317
McDonald JW, Silverstein FS, Johnston MV (1988) Neurotoxicity of N-methyl-D-aspartate is markedly enhanced in developing rat central nervous system. Brain Res 459: 200–203
Ikonomidou C, Mosinger JL, Salles KS et al (1989) Sensitivity of the developing rat brain to hypobaric/ischemic damage parallels sensitivity to N-methyl-aspartate neurotoxicity. J Neurosci 9: 2809–2818
West T, Atzeva M, Holtzman DM (2006) Caspase-3 deficiency during development increases vulnerability to hypoxic-ischemic injury through caspase-3-independent pathways. Neurobiol Dis 22: 523–537
Parsadanian AS, Cheng Y, Keller-Peck CR et al (1998) Bcl-xL is an antiapoptotic regulator for postnatal CNS neurons. J Neurosci 18: 1009–1019
Kawane K, Fukuyama H, Yoshida H et al (2003) Impaired thymic development in mouse embryos deficient in apoptotic DNA degradation. Nat Immunol 4: 138–144
Kuida K, Zheng TS, Na S et al (1996) Decreased apoptosis in the brain and premature lethality in CPP32-deficient mice. Nature 384: 368–372
Lakhani SA, Masud A, Kuida K et al (2006) Caspases 3 and 7: key mediators of mitochondrial events of apoptosis. Science 311: 847–851
Leonard JR, Klocke BJ, D’Sa C et al (2002) Strain-dependent neurodevelopmental abnormalities in caspase-3-deficient mice. J Neuropathol Exp Neurol 61: 673–677
Reznikov KY (1991) Cell proliferation and cytogenesis in the mouse hippocampus. Adv Anat Embryol Cell Biol 122: 1–74
Houde C, Banks KG, Coulombe N et al (2004) Caspase-7 expanded function and intrinsic expression level underlies strain-specific brain phenotype of caspase-3-null mice. J Neurosci 24: 9977–9984
Clarke PG (1990) Developmental cell death: morphological diversity and multiple mechanisms. Anat Embryol (Berl) 181: 195–213
Bursch W (2001) The autophagosomal-lysosomal compartment in programmed cell death. Cell Death Differ 8: 569–581
Canu N, Tufi R, Serafino AL et al (2005) Role of the autophagic-lysosomal system on low potassium-induced apoptosis in cultured cerebellar granule cells. J Neurochem 92: 1228–1242
Isahara K, Ohsawa Y, Kanamori S et al (1999) Regulation of a novel pathway for cell death by lysosomal aspartic and cysteine proteinases. Neuroscience 91: 233–249
Ohsawa Y, Isahara K, Kanamori S et al (1998) An ultrastructural and immunohistochemical study of PC12 cells during apoptosis induced by serum deprivation with special reference to autophagy and lysosomal cathepsins. Arch Histol Cytol 61: 395–403
Shibata M, Kanamori S, Isahara K et al (1998) Participation of cathepsins B and D in apoptosis of PC12 cells following serum deprivation. Biochem Biophys Res Commun 251: 199–203
Telbisz A, Kovacs AL, Somosy Z (2002) Influence of X-ray on the autophagic-lysosomal system in rat pancreatic acini. Micron 33: 143–151
Uchiyama Y (2001) Autophagic cell death and its execution by lysosomal cathepsins. Arch Histol Cytol 64: 233–246
Yu L, Alva A, Su H et al (2004) Regulation of an ATG7-beclin 1 program of autophagic cell death by caspase-8. Science 304: 1500–1502
Bursch W, Ellinger A, Kienzl H et al (1996) Active cell death induced by the anti-estrogens tamoxifen and ICI 164 384 in human mammary carcinoma cells (MCF-7) in culture: the role of autophagy. Carcinogenesis 17: 1595–1607
Ploeg RJ, D’Alessandro AM, Knechtle SJ et al (1993) Risk factors for primary dysfunction after liver transplantation—a multivariate analysis. Transplantation 55: 807–813
Strasberg SM, Howard TK, Molmenti EP et al (1994) Selecting the donor liver: risk factors for poor function after orthotopic liver transplantation. Hepatology 20: 829–838
Calmus Y, Cynober L, Dousset B et al (1995) Evidence for the detrimental role of proteolysis during liver preservation in humans. Gastroenterology 108: 1510–1516
Furukawa H, Todo S, Imventarza O et al (1991) Effect of cold ischemia time on the early outcome of human hepatic allografts preserved with UW solution. Transplantation 51: 1000–1004
Gotoh K, Lu Z, Morita M et al (2009) Participation of autophagy in the initiation of graft dysfunction after rat liver transplantation. Autophagy 5: 351–360
Lu Z, Dono K, Gotoh K et al (2005) Participation of autophagy in the degeneration process of rat hepatocytes after transplantation following prolonged cold preservation. Arch Histol Cytol 68: 71–80
Cheng Y, Deshmukh M, D’Costa A et al (1998) Caspase inhibitor affords neuroprotection with delayed administration in a rat model of neonatal hypoxic-ischemic brain injury. J Clin Invest 101: 1992–1999
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2010 Springer
About this paper
Cite this paper
Uchiyama, Y., Koike, M., Shibata, M. (2010). Cell Death and Autophagy. In: Tamaki, N., Kuge, Y. (eds) Molecular Imaging for Integrated Medical Therapy and Drug Development. Springer, Tokyo. https://doi.org/10.1007/978-4-431-98074-2_19
Download citation
DOI: https://doi.org/10.1007/978-4-431-98074-2_19
Publisher Name: Springer, Tokyo
Print ISBN: 978-4-431-98073-5
Online ISBN: 978-4-431-98074-2
eBook Packages: MedicineMedicine (R0)