title: Acetylcholine receptors from human muscle as pharmacological targets for ALS therapy.
abstract: Amyotrophic lateral sclerosis (ALS) is a neurodegenerative disease affecting motor neurons that leads to progressive paralysis of skeletal muscle. Studies of ALS have revealed defects in expression of acetylcholine receptors (AChRs) in skeletal muscle that occur even in the absence of motor neuron anomalies. The endocannabinoid palmitoylethanolamide (PEA) modified the clinical conditions in one ALS patient, improving muscle force and respiratory efficacy. By microtransplanting muscle membranes from selected ALS patients into Xenopus oocytes, we show that PEA reduces the desensitization of acetylcholine-evoked currents after repetitive neurotransmitter application (i.e., rundown). The same effect was observed using muscle samples from denervated (non-ALS) control patients. The expression of human recombinant α1β1γδ (γ-AChRs) and α1β1εδ AChRs (ε-AChRs) in Xenopus oocytes revealed that PEA selectively affected the rundown of ACh currents in ε-AChRs. A clear up-regulation of the α1 subunit in muscle from ALS patients compared with that from non-ALS patients was found by quantitative PCR, but no differential expression was found for other subunits. Clinically, ALS patients treated with PEA showed a lower decrease in their forced vital capacity (FVC) over time as compared with untreated ALS patients, suggesting that PEA can enhance pulmonary function in ALS. In the present work, data were collected from a cohort of 76 ALS patients and 17 denervated patients. Our results strengthen the evidence for the role of skeletal muscle in ALS pathogenesis and pave the way for the development of new drugs to hamper the clinical effects of the disease.
authors.affiliation: Department of Physiology and Pharmacology, Istituto Pasteur-Fondazione Cenci Bolognetti, University of Rome Sapienza, 00185 Rome, Italy; Istituto di Ricovero e Cura a Carattere Scientifico San Raffaele Pisana, 00166 Rome, Italy; eleonora.palma@uniroma1.it rmiledi@uci.edu.
authors.forename: Eleonora
authors.initials: E
authors.lastname: Palma
authors.affiliation: Department of Neurobiology and Behavior, University of California, Irvine, CA 92697;
authors.forename: Jorge Mauricio
authors.initials: JM
authors.lastname: Reyes-Ruiz
authors.affiliation: Department of Neurology and Psychiatry, University of Rome Sapienza, 00185 Rome, Italy;
authors.forename: Diego
authors.initials: D
authors.lastname: Lopergolo
authors.affiliation: Istituto di Ricovero e Cura a Carattere Scientifico San Raffaele Pisana, 00166 Rome, Italy;
authors.forename: Cristina
authors.initials: C
authors.lastname: Roseti
authors.affiliation: Department of Physiology and Pharmacology, Istituto Pasteur-Fondazione Cenci Bolognetti, University of Rome Sapienza, 00185 Rome, Italy;
authors.forename: Cristina
authors.initials: C
authors.lastname: Bertollini
authors.affiliation: Department of Physiology and Pharmacology, Istituto Pasteur-Fondazione Cenci Bolognetti, University of Rome Sapienza, 00185 Rome, Italy;
authors.forename: Gabriele
authors.initials: G
authors.lastname: Ruffolo
authors.affiliation: Department of Physiology and Pharmacology, Istituto Pasteur-Fondazione Cenci Bolognetti, University of Rome Sapienza, 00185 Rome, Italy; Ri.MED Foundation, 90133 Palermo, Italy;
authors.forename: Pierangelo
authors.initials: P
authors.lastname: Cifelli
authors.affiliation: Department of Neurology and Psychiatry, University of Rome Sapienza, 00185 Rome, Italy;
authors.forename: Emanuela
authors.initials: E
authors.lastname: Onesti
authors.affiliation: Department of Physiology and Pharmacology, Istituto Pasteur-Fondazione Cenci Bolognetti, University of Rome Sapienza, 00185 Rome, Italy; Istituto di Ricovero e Cura a Carattere Scientifico Neuromed, 86077 Pozzilli, Isernia, Italy;
authors.forename: Cristina
authors.initials: C
authors.lastname: Limatola
authors.affiliation: Department of Neurobiology and Behavior, University of California, Irvine, CA 92697; Instituto de Neurobiología, Universidad Nacional Autónoma de México, Juriquilla, 76230 Querétaro, México eleonora.palma@uniroma1.it rmiledi@uci.edu.
authors.forename: Ricardo
authors.initials: R
authors.lastname: Miledi
authors.affiliation: Department of Neurology and Psychiatry, University of Rome Sapienza, 00185 Rome, Italy;
authors.forename: Maurizio
authors.initials: M
authors.lastname: Inghilleri
mesh_terms: D000200:Action Potentials / Q000187:drug effects; D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D000577:Amides; D000690:Amyotrophic Lateral Sclerosis / Q000188:drug therapy* / Q000473:pathology / Q000503:physiopathology; D000818:Animals; D002462:Cell Membrane / Q000187:drug effects / Q000637:transplantation; D004983:Ethanolamines / Q000494:pharmacology / Q000627:therapeutic use*; D005260:Female; D006801:Humans; D008297:Male; D008845:Microinjections; D008875:Middle Aged; D058990:Molecular Targeted Therapy; D009121:Muscle Denervation; D018482:Muscle, Skeletal / Q000187:drug effects* / Q000648:ultrastructure; D009469:Neuromuscular Junction / Q000503:physiopathology; D009865:Oocytes; D010169:Palmitic Acids / Q000494:pharmacology / Q000627:therapeutic use*; D011978:Receptors, Nicotinic / Q000187:drug effects* / Q000502:physiology; D011993:Recombinant Fusion Proteins / Q000187:drug effects / Q000235:genetics; D016037:Single-Blind Method; D014982:Xenopus laevis
chemical_list: D000577:Amides; C504423:CHRNE protein, human; C539275:CHRNG protein, human; D004983:Ethanolamines; D010169:Palmitic Acids; D011978:Receptors, Nicotinic; D011993:Recombinant Fusion Proteins; C005958:palmidrol
keywords: ALS; Xenopus oocytes; microtransplantation; muscle AChR; qPCR