Abstract
Autophagy is a regulated lysosomal degradation process that involves autophagosome formation and transport. Although recent evidence indicates that basal levels of autophagy protect against neurodegeneration, the exact mechanism whereby this occurs is not known. By using conditional knockout mutant mice, we report that neuronal autophagy is particularly important for the maintenance of local homeostasis of axon terminals and protection against axonal degeneration. We show that specific ablation of an essential autophagy gene, Atg7, in Purkinje cells initially causes cell-autonomous, progressive dystrophy (manifested by axonal swellings) and degeneration of the axon terminals. Consistent with suppression of autophagy, no autophagosomes are observed in these dystrophic swellings, which is in contrast to accumulation of autophagosomes in the axonal dystrophic swellings under pathological conditions. Axonal dystrophy of mutant Purkinje cells proceeds with little sign of dendritic or spine atrophy, indicating that axon terminals are much more vulnerable to autophagy impairment than dendrites. This early pathological event in the axons is followed by cell-autonomous Purkinje cell death and mouse behavioral deficits. Furthermore, ultrastructural analyses of mutant Purkinje cells reveal an accumulation of aberrant membrane structures in the axonal dystrophic swellings. Finally, we observe double-membrane vacuole-like structures in wild-type Purkinje cell axons, whereas these structures are abolished in mutant Purkinje cell axons. Thus, we conclude that the autophagy protein Atg7 is required for membrane trafficking and turnover in the axons. Our study implicates impairment of axonal autophagy as a possible mechanism for axonopathy associated with neurodegeneration.
MeSH Terms
Adaptor Proteins, Signal Transducing/metabolism
Animals
Autophagy
Autophagy-Related Protein 7
Axons/metabolism
Cell Membrane/metabolism
Cell Shape
Genes, Reporter/genetics
Guanine Nucleotide Exchange Factors/metabolism
Heat-Shock Proteins/metabolism
Homeostasis
Mice
Mice, Transgenic
Microscopy, Electron, Transmission
Microtubule-Associated Proteins/genetics,metabolism
Nerve Degeneration/genetics,metabolism,pathology,prevention & control
Neuropeptides/metabolism
Purkinje Cells/cytology,metabolism
Sequestosome-1 Protein
Spine/cytology,metabolism
Chemicals
Adaptor Proteins, Signal Transducing
Atg7 protein, mouse
Guanine Nucleotide Exchange Factors
Heat-Shock Proteins
Microtubule-Associated Proteins
Neuropeptides
Pcp2 protein, mouse
Sequestosome-1 Protein
Sqstm1 protein, mouse
Autophagy-Related Protein 7
Authors & Affiliations
9 authors, click to expand affiliations / ORCID
Komatsu Masaaki
Departments of Neurology and Neuroscience, Mount Sinai School of Medicine, New York, NY 10029, USA.
Wang Qing Jun
Holstein Gay R
Friedrich Victor L
Iwata Jun-ichi
Kominami Eiki
Chait Brian T
Tanaka Keiji
Yue Zhenyu
References (31)
31 references, click to expand
-
LC3, a mammalian homologue of yeast Apg8p, is localized in autophagosome membranes after processing.
EMBO J. 2000 Nov 1;19(21):5720-8
PMID: 11060023
-
Induction of autophagy in axonal dystrophy and degeneration.
J Neurosci. 2006 Aug 2;26(31):8057-68
PMID: 16885219
-
Huntingtin aggregate-associated axonal degeneration is an early pathological event in Huntington's disease mice.
J Neurosci. 2001 Nov 1;21(21):8473-81
PMID: 11606636
-
Axon pathology in neurological disease: a neglected therapeutic target.
Trends Neurosci. 2002 Oct;25(10):532-7
PMID: 12220882
-
A novel protein complex linking the delta 2 glutamate receptor and autophagy: implications for neurodegeneration in lurcher mice.
Neuron. 2002 Aug 29;35(5):921-33
PMID: 12372286
-
In vivo analysis of autophagy in response to nutrient starvation using transgenic mice expressing a fluorescent autophagosome marker.
Mol Biol Cell. 2004 Mar;15(3):1101-11
PMID: 14699058
-
Ultrastructural neuronal pathology in transgenic mice expressing mutant (P301L) human tau.
J Neurocytol. 2003 Nov;32(9):1091-105
PMID: 15044841
-
Development by self-digestion: molecular mechanisms and biological functions of autophagy.
Dev Cell. 2004 Apr;6(4):463-77
PMID: 15068787
-
Two ubiquitin-like conjugation systems essential for autophagy.
Semin Cell Dev Biol. 2004 Apr;15(2):231-6
PMID: 15209383
-
Autophagy is a part of ultrastructural synaptic pathology in Creutzfeldt-Jakob disease: a brain biopsy study.
Int J Biochem Cell Biol. 2004 Dec;36(12):2563-73
PMID: 15325593
-
"Phagocytic" lysosomes in chromatolytic neurones.
Nature. 1967 Aug 5;215(5101):657-8
PMID: 6050233
-
A light and electron microscopic study of the cellular response to axonal injury in the superior cervical ganglion of the rat.
Proc R Soc Lond B Biol Sci. 1972 Apr 18;181(1062):43-79
PMID: 4402334
-
Electron microscope and enzyme studies in cerebral biopsies of Huntington's chorea.
Trans Am Neurol Assoc. 1974;99:240-3
PMID: 4282536
-
The neuronal endoplasmic reticulum: its cytochemistry and contribution to the endomembrane system. II. Axons and terminals.
J Comp Neurol. 1984 Dec 1;230(2):231-48
PMID: 6210310
-
Products of endocytosis and autophagy are retrieved from axons by regulated retrograde organelle transport.
J Cell Biol. 1993 Apr;121(2):305-15
PMID: 7682217
-
Properties of the endosomal-lysosomal system in the human central nervous system: disturbances mark most neurons in populations at risk to degenerate in Alzheimer's disease.
J Neurosci. 1996 Jan;16(1):186-99
PMID: 8613784
-
Apoptosis and autophagy in nigral neurons of patients with Parkinson's disease.
Histol Histopathol. 1997 Jan;12(1):25-31
PMID: 9046040
-
Endocytotic formation of vesicles and other membranous structures induced by Ca2+ and axolemmal injury.
J Neurosci. 1998 Jun 1;18(11):4029-41
PMID: 9592084
-
Conformational changes of the smooth endoplasmic reticulum are facilitated by L-glutamate and its receptors in rat Purkinje cells.
J Comp Neurol. 1998 Dec 14;402(2):252-63
PMID: 9845247
-
Extensive involvement of autophagy in Alzheimer disease: an immuno-electron microscopy study.
J Neuropathol Exp Neurol. 2005 Feb;64(2):113-22
PMID: 15751225
-
Electron tomography of degenerating neurons in mice with abnormal regulation of iron metabolism.
J Struct Biol. 2005 May;150(2):144-53
PMID: 15866737
-
Impairment of starvation-induced and constitutive autophagy in Atg7-deficient mice.
J Cell Biol. 2005 May 9;169(3):425-34
PMID: 15866887
-
Macroautophagy--a novel Beta-amyloid peptide-generating pathway activated in Alzheimer's disease.
J Cell Biol. 2005 Oct 10;171(1):87-98
PMID: 16203860
-
Subsurface cisterna-lined axonal invaginations and double-walled vesicles at the axonal-myelin sheath interface.
Neurosci Res. 2005 Nov;53(3):298-303
PMID: 16129504
-
Axon degeneration mechanisms: commonality amid diversity.
Nat Rev Neurosci. 2005 Nov;6(11):889-98
PMID: 16224497
-
p62/SQSTM1 forms protein aggregates degraded by autophagy and has a protective effect on huntingtin-induced cell death.
J Cell Biol. 2005 Nov 21;171(4):603-14
PMID: 16286508
-
Generation of cell lines with tetracycline-regulated autophagy and a role for autophagy in controlling cell size.
FEBS Lett. 2006 May 15;580(11):2623-9
PMID: 16647067
-
Suppression of basal autophagy in neural cells causes neurodegenerative disease in mice.
Nature. 2006 Jun 15;441(7095):885-9
PMID: 16625204
-
Loss of autophagy in the central nervous system causes neurodegeneration in mice.
Nature. 2006 Jun 15;441(7095):880-4
PMID: 16625205
-
Autophagy and its possible roles in nervous system diseases, damage and repair.
Autophagy. 2005 Apr;1(1):11-22
PMID: 16874045
-
Cre recombinase expression in cerebellar Purkinje cells.
Genesis. 2000 Nov-Dec;28(3-4):93-8
PMID: 11105049