title: VGF Protein and Its C-Terminal Derived Peptides in Amyotrophic Lateral Sclerosis: Human and Animal Model Studies.
abstract: VGF mRNA is widely expressed in areas of the nervous system known to degenerate in Amyotrophic Lateral Sclerosis (ALS), including cerebral cortex, brainstem and spinal cord. Despite certain VGF alterations are reported in animal models, little information is available with respect to the ALS patients. We addressed VGF peptide changes in fibroblast cell cultures and in plasma obtained from ALS patients, in parallel with spinal cord and plasma samples from the G93A-SOD1 mouse model. Antisera specific for the C-terminal end of the human and mouse VGF proteins, respectively, were used in immunohistochemistry and enzyme-linked immunosorbent assay (ELISA), while gel chromatography and HPLC/ESI-MS/MS were used to identify the VGF peptides present. Immunoreactive VGF C-terminus peptides were reduced in both fibroblast and plasma samples from ALS patients in an advanced stage of the disease. In the G93A-SOD1 mice, the same VGF peptides were also decreased in plasma in the late-symptomatic stage, while showing an earlier down-regulation in the spinal cord. In immunohistochemistry, a large number of gray matter structures were VGF C-terminus immunoreactive in control mice (including nerve terminals, axons and a few perikarya identified as motoneurons), with a striking reduction already in the pre-symptomatic stage. Through gel chromatography and spectrometry analysis, we identified one form likely to be the VGF precursor as well as peptides containing the NAPP- sequence in all tissues studied, while in the mice and fibroblasts, we revealed also AQEE- and TLQP- peptides. Taken together, selective VGF fragment depletion may participate in disease onset and/or progression of ALS.
authors.affiliation: NEF-Laboratory, Department of Biomedical Sciences, University of Cagliari, Cagliari, Italy.
authors.forename: Carla
authors.identifier: http://orcid.org/0000-0002-0716-3983
authors.initials: C
authors.lastname: Brancia
authors.affiliation: NEF-Laboratory, Department of Biomedical Sciences, University of Cagliari, Cagliari, Italy.
authors.forename: Barbara
authors.initials: B
authors.lastname: Noli
authors.affiliation: Neuroscience Institute Cavalieri Ottolenghi, Department of Neuroscience, University of Turin, Turin, Italy.
authors.forename: Marina
authors.initials: M
authors.lastname: Boido
authors.affiliation: NEF-Laboratory, Department of Biomedical Sciences, University of Cagliari, Cagliari, Italy.
authors.forename: Andrea
authors.initials: A
authors.lastname: Boi
authors.affiliation: Department of Neurology, Azienda Universitario Ospedaliera di Cagliari and University of Cagliari, Cagliari, Italy.
authors.forename: Roberta
authors.initials: R
authors.lastname: Puddu
authors.affiliation: Department of Neurology, Azienda Universitario Ospedaliera di Cagliari and University of Cagliari, Cagliari, Italy.
authors.forename: Giuseppe
authors.initials: G
authors.lastname: Borghero
authors.affiliation: Department of Neurology, Azienda Universitario Ospedaliera di Cagliari and University of Cagliari, Cagliari, Italy.
authors.forename: Francesco
authors.initials: F
authors.lastname: Marrosu
authors.affiliation: Neurorehabilitation Unit, Neuroscience Department, University of Pisa, Pisa, Italy.
authors.forename: Paolo
authors.initials: P
authors.lastname: Bongioanni
authors.affiliation: Medical Genetics, Department of Medical Sciences, University of Cagliari, Binaghi Hospital, Cagliari, Italy.
authors.forename: Sandro
authors.initials: S
authors.lastname: Orrù
authors.affiliation: Department of Life and Environmental Sciences, University of Cagliari, Cagliari, Italy.
authors.forename: Barbara
authors.initials: B
authors.lastname: Manconi
authors.affiliation: NEF-Laboratory, Department of Biomedical Sciences, University of Cagliari, Cagliari, Italy.
authors.forename: Filomena
authors.initials: F
authors.lastname: D'Amato
authors.affiliation: Institute of Chemistry of the Molecular Recognition CNR, Rome, Italy.
authors.forename: Irene
authors.initials: I
authors.lastname: Messana
authors.affiliation: Institute of Biochemistry and Clinical Biochemistry, Catholic University, Rome, Italy.
authors.forename: Federica
authors.initials: F
authors.lastname: Vincenzoni
authors.affiliation: Neuroscience Institute Cavalieri Ottolenghi, Department of Neuroscience, University of Turin, Turin, Italy.
authors.forename: Alessandro
authors.initials: A
authors.lastname: Vercelli
authors.affiliation: NEF-Laboratory, Department of Biomedical Sciences, University of Cagliari, Cagliari, Italy.
authors.forename: Gian-Luca
authors.initials: GL
authors.lastname: Ferri
authors.affiliation: NEF-Laboratory, Department of Biomedical Sciences, University of Cagliari, Cagliari, Italy.
authors.forename: Cristina
authors.initials: C
authors.lastname: Cocco
mesh_terms: D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D000690:Amyotrophic Lateral Sclerosis / Q000378:metabolism / Q000473:pathology*; D000818:Animals; D002478:Cells, Cultured; D002851:Chromatography, High Pressure Liquid; D004195:Disease Models, Animal; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D005347:Fibroblasts / Q000166:cytology / Q000378:metabolism; D006801:Humans; D007150:Immunohistochemistry; D008297:Male; D051379:Mice; D008822:Mice, Transgenic; D008875:Middle Aged; D009414:Nerve Growth Factors / Q000032:analysis / Q000097:blood*; D009479:Neuropeptides / Q000032:analysis / Q000097:blood*; D021241:Spectrometry, Mass, Electrospray Ionization; D013116:Spinal Cord / Q000378:metabolism; D013482:Superoxide Dismutase / Q000235:genetics / Q000378:metabolism
chemical_list: D009414:Nerve Growth Factors; D009479:Neuropeptides; C522413:VGF peptide; C510637:VGF protein, human; D013482:Superoxide Dismutase