title: Proteomic and histochemical analysis of proteins involved in the dying-back-type of axonal degeneration in the gracile axonal dystrophy (gad) mouse.
abstract: Local axonal degeneration is a common pathological feature of peripheral neuropathies and neurodegenerative disorders of the central nervous system, including Alzheimer's disease, Parkinson's disease, and stroke; however, the underlying molecular mechanism is not known. Here, we analyzed the gracile axonal dystrophy (gad) mouse, which displays the dying-back-type of axonal degeneration in sensory neurons, to find the molecules involved in the mechanism of axonal degeneration. The gad mouse is analogous to a null mutant of ubiquitin carboxyl-terminal hydrolase L1 (UCH-L1). UCH-L1 is a deubiquitinating enzyme expressed at high levels in neurons, as well as testis and ovary. In addition, we recently discovered a new function of UCH-L1-namely to bind to and stabilize mono-ubiquitin in neurons, and found that the level of mono-ubiquitin was decreased in neurons, especially in axons of the sciatic nerve, in gad mice. The low level of ubiquitin suggests that the target proteins of the ubiquitin proteasome system are not sufficiently ubiquitinated and thus degraded in the gad mouse; therefore, these proteins may be the key molecules involved in axonal degeneration. To identify molecules involved in axonal degeneration in gad mice, we compared protein expression in sciatic nerves between gad and wild-type mice at 2 and 12 weeks old, using two-dimensional difference gel electrophoresis. As a result, we found age-dependent accumulation of several proteins, including glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and 14-3-3, in gad mice compared with wild-type mice. Histochemical analyses demonstrated that GAPDH and 14-3-3 were localized throughout axons in both gad and wild-type mice, but GAPDH accumulated in the axons of gad mice. Recently, it has been suggested that a wide range of neurodegenerative diseases are characterized by the accumulation of intracellular and extracellular protein aggregates, and it has been reported that oxidative stress causes the aggregation of GAPDH. Furthermore, histochemical analysis demonstrated that sulfonated GAPDH, a sensor of oxidative stress that elicits cellular dysfunction, was expressed in the axons of gad mice, and 4-hydroxy-2-nonenal, a major marker of oxidative stress, was also only detected in gad mice. Our findings suggest that GAPDH may participate in a process of the dying-back-type of axonal degeneration in gad mice and may provide valuable insight into the mechanisms of axonal degeneration.
authors.affiliation: Department of Degenerative Neurological Diseases, National Institute of Neuroscience, National Center of Neurology and Psychiatry, Ogawahigashi, Kodaira, Tokyo, Japan.
authors.forename: Akiko
authors.initials: A
authors.lastname: Goto
authors.forename: Yu-Lai
authors.initials: YL
authors.lastname: Wang
authors.forename: Tomohiro
authors.initials: T
authors.lastname: Kabuta
authors.forename: Rieko
authors.initials: R
authors.lastname: Setsuie
authors.forename: Hitoshi
authors.initials: H
authors.lastname: Osaka
authors.forename: Akira
authors.initials: A
authors.lastname: Sawa
authors.forename: Shoichi
authors.initials: S
authors.lastname: Ishiura
authors.forename: Keiji
authors.initials: K
authors.lastname: Wada
mesh_terms: D048948:14-3-3 Proteins / Q000032:analysis / Q000378:metabolism; D000375:Aging / Q000378:metabolism / Q000473:pathology; D000447:Aldehydes / Q000378:metabolism; D000818:Animals; D001369:Axons / Q000378:metabolism* / Q000473:pathology; D015415:Biomarkers / Q000032:analysis / Q000378:metabolism; D004195:Disease Models, Animal; D015180:Electrophoresis, Gel, Two-Dimensional; D024601:Glyceraldehyde 3-Phosphate Dehydrogenase (NADP+) / Q000032:analysis / Q000378:metabolism*; D006651:Histocytochemistry; D051379:Mice; D008818:Mice, Neurologic Mutants; D018384:Oxidative Stress / Q000502:physiology; D040901:Proteomics; D012584:Sciatic Nerve / Q000378:metabolism* / Q000473:pathology / Q000503:physiopathology; D011984:Sensory Receptor Cells / Q000378:metabolism* / Q000473:pathology; D014855:Wallerian Degeneration / Q000235:genetics / Q000378:metabolism* / Q000503:physiopathology
chemical_list: D048948:14-3-3 Proteins; D000447:Aldehydes; D015415:Biomarkers; D024601:Glyceraldehyde 3-Phosphate Dehydrogenase (NADP+); C027576:4-hydroxy-2-nonenal