title: Immunoreactivity of valosin-containing protein in sporadic amyotrophic lateral sclerosis and in a case of its novel mutant.
abstract: BACKGROUND
  Mutations in the valosin-containing protein (VCP) gene were first found to cause inclusion- body myopathy with early-onset Paget disease and frontotemporal dementia (IBMPFD). Mutations in the VCP gene were later reported to occur in familial amyotrophic lateral sclerosis (ALS). But the role of VCP in the neurodegenerative processes that occur in ALS remains unknown. The purpose of the present study was to elucidate the role of VCP in the neurodegeneration seen in sporadic and VCP mutant ALS.
  
  
  RESULTS
  Immunohistochemistry demonstrated that the frequency of distinct VCP-positive nuclei of spinal motor neurons of patients with sporadic ALS (SALS) and the ALS with VCP novel mutation (ALS-VCP, M158V) was increased, compared with that of the control cases. No VCP-positive inclusion bodies were observed in SALS patients, a ALS-VCP patient or in control subjects. Neuropathologic examination of the ALS-VCP case showed loss of motor neurons, the presence of Bunina bodies, and degeneration of the corticospinal tracts. Bunina bodies detected in this case were confirmed to show immunohistochemical and ultrastructural features similar to those previously described. Furthermore, neuronal intracytoplasmic inclusions immunopositive for TAR DNA-binding protein 43 kDa (TDP-43), phosphorylated TDP-43, ubiquitin (Ub), p62, and optineurin were identified in the spinal and medullary motoneurons, but not in the neocortex. Gene analysis of this ALS-VCP patient confirmed the de novo mutation of M158V, which was not found in control cases; and bioinformatics using several in silico analyses showed possible damage to the structure of VCP. Immunocytochemical study of cultured cells showed increased cytoplasmic translocation of TDP-43 in cells transfected with several mutant VCP including our patient's compared with wild-type VCP.
  
  
  CONCLUSIONS
  These findings support the idea that VCP is associated with the pathomechanism of SALS and familial ALS with a VCP mutation, presumably acting through a dominant-negative mechanism.
authors.affiliation: Department of Neurology, Wakayama Medical University, 811-1, Kimiidera, Wakayama, 641-8510, Japan. ayaki520@kuhp.kyoto-u.ac.jp.
authors.forename: Takashi
authors.initials: T
authors.lastname: Ayaki
authors.affiliation: Department of Neurology, Wakayama Medical University, 811-1, Kimiidera, Wakayama, 641-8510, Japan. ito@wakayama-med.ac.jp.
authors.forename: Hidefumi
authors.initials: H
authors.lastname: Ito
authors.affiliation: Department of Pathology, Osaka City General Hospital, 2-13-22, Miyakojima-Hondori, Miyakojima-ku, Osaka, 534-0021, Japan. hir-fukushima@med.osakacity-hp.or.jp.
authors.forename: Hiroko
authors.initials: H
authors.lastname: Fukushima
authors.affiliation: Department of Pathology, Osaka City General Hospital, 2-13-22, Miyakojima-Hondori, Miyakojima-ku, Osaka, 534-0021, Japan. m1657676@msic.med.osaka-cu.ac.jp.
authors.forename: Takeshi
authors.initials: T
authors.lastname: Inoue
authors.affiliation: Center for iPS Cell Research and Application, Kyoto University, 53, Kawahara-cho, Shogoin, Sakyo-ku, Kyoto, 606-8507, Japan. kondotak@kuhp.kyoto-u.ac.jp.|Department of Neurology, Kyoto University Graduate School of Medicine, 53, Kawahara-cho, Shogoin, Sakyo-ku, Kyoto, 606-8507, Japan. kondotak@kuhp.kyoto-u.ac.jp.
authors.forename: Takayuki
authors.initials: T
authors.lastname: Kondo
authors.affiliation: Kyoto Municipal Rehabilitation Center for Physically Disabled People, 30, Mibusennen-cho, Nakagyo-ku,, Kyoto, 604-8854, Japan. akito@kuhp.kyoto-u.ac.jp.
authors.forename: Akito
authors.initials: A
authors.lastname: Ikemoto
authors.affiliation: Department of Neurology, Kyoto University Graduate School of Medicine, 53, Kawahara-cho, Shogoin, Sakyo-ku, Kyoto, 606-8507, Japan. tasano@kuhp.kyoto-u.ac.jp.
authors.forename: Takeshi
authors.initials: T
authors.lastname: Asano
authors.affiliation: Department of Neurology, Kyoto University Graduate School of Medicine, 53, Kawahara-cho, Shogoin, Sakyo-ku, Kyoto, 606-8507, Japan. shodai@kuhp.kyoto-u.ac.jp.
authors.forename: Akemi
authors.initials: A
authors.lastname: Shodai
authors.affiliation: Takumi Medical Corporation, Neurology Clinic, 7-8, Takarayama-cho, Toyonaka, Osaka, 561-0893, Japan. fujita-okhc@healthcarenet.jp.
authors.forename: Takuji
authors.initials: T
authors.lastname: Fujita
authors.affiliation: Tazuke Kofukai Foundation, Medical Research Institute and Kitano Hospital, 2-4-20, Ohgimachi, Kita-ku, Osaka, 530-8480, Japan. sfukui@kitano-hp.or.jp.
authors.forename: Satoshi
authors.initials: S
authors.lastname: Fukui
authors.affiliation: Department of Epidemiology, Research Institute for Radiation Biology and Medicine, Hiroshima University, 1-2-3, Kasumi, Minami-ku, Hiroshima, 734-8553, Japan. morino@hiroshima-u.ac.jp.
authors.forename: Hiroyuki
authors.initials: H
authors.lastname: Morino
authors.affiliation: Department of Neurology, Osaka City General Hospital, 2-13-22, Miyakojima-Hondori, Miyakojima-ku, Osaka, 534-0021, Japan. s-nakano@med.osakacity-hp.or.jp.
authors.forename: Satoshi
authors.initials: S
authors.lastname: Nakano
authors.affiliation: Department of Neurology, Kansai Medical University, 2-5-1, Shin-machi, Hirakata, Osaka, 573-1010, Japan. kusaka@hirakata.kmu.ac.jp.
authors.forename: Hirofumi
authors.initials: H
authors.lastname: Kusaka
authors.affiliation: Department of Neurology, Kyoto University Graduate School of Medicine, 53, Kawahara-cho, Shogoin, Sakyo-ku, Kyoto, 606-8507, Japan. hirofumi@kuhp.kyoto-u.ac.jp.
authors.forename: Hirofumi
authors.initials: H
authors.lastname: Yamashita
authors.affiliation: Department of Stroke and Cerebrovascular Diseases, National Cerebral and Cardiovascular Center, 5-7-1, Fujishirodai, Suita, Osaka, 565-8565, Japan. ihara@kuhp.kyoto-u.ac.jp.
authors.forename: Masafumi
authors.initials: M
authors.lastname: Ihara
authors.affiliation: Department of Neurology, Kyoto University Graduate School of Medicine, 53, Kawahara-cho, Shogoin, Sakyo-ku, Kyoto, 606-8507, Japan. matsumot@kuhp.kyoto-u.ac.jp.|Department of Epilepsy, Movement Disorders and Physiology, Kyoto University Graduate School of Medicine, 53, Kawahara-cho, Shogoin, Sakyo-ku, Kyoto, 606-8507, Japan. matsumot@kuhp.kyoto-u.ac.jp.
authors.forename: Riki
authors.initials: R
authors.lastname: Matsumoto
authors.affiliation: Department of Neurology, School of Medicine, Sapporo Medical University, South 1 West 17, Chuo-ku, Sapporo, Hokkaido, 060-8556, Japan. kawamata@kuhp.kyoto-u.ac.jp.
authors.forename: Jun
authors.initials: J
authors.lastname: Kawamata
authors.affiliation: Department of Neurology, Kyoto University Graduate School of Medicine, 53, Kawahara-cho, Shogoin, Sakyo-ku, Kyoto, 606-8507, Japan. urumak@kuhp.kyoto-u.ac.jp.
authors.forename: Makoto
authors.initials: M
authors.lastname: Urushitani
authors.affiliation: Department of Epidemiology, Research Institute for Radiation Biology and Medicine, Hiroshima University, 1-2-3, Kasumi, Minami-ku, Hiroshima, 734-8553, Japan. kawakamihideshi@gmail.com.
authors.forename: Hideshi
authors.initials: H
authors.lastname: Kawakami
authors.affiliation: Department of Neurology, Kyoto University Graduate School of Medicine, 53, Kawahara-cho, Shogoin, Sakyo-ku, Kyoto, 606-8507, Japan. ryosuket@kuhp.kyoto-u.ac.jp.
authors.forename: Ryosuke
authors.initials: R
authors.lastname: Takahashi
mesh_terms: D000251:Adenosine Triphosphatases / Q000235:genetics*; D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D000690:Amyotrophic Lateral Sclerosis / Q000235:genetics* / Q000378:metabolism* / Q000473:pathology; D001921:Brain / Q000378:metabolism* / Q000473:pathology; D018797:Cell Cycle Proteins / Q000235:genetics*; D045744:Cell Line, Tumor; D005260:Female; D057809:HEK293 Cells; D006801:Humans; D008297:Male; D008875:Middle Aged; D009154:Mutation; D013116:Spinal Cord / Q000378:metabolism* / Q000473:pathology; D000074405:Valosin Containing Protein
chemical_list: D018797:Cell Cycle Proteins; D000251:Adenosine Triphosphatases; C000614178:VCP protein, human; D000074405:Valosin Containing Protein