title: Protein-anchoring strategy for delivering acetylcholinesterase to the neuromuscular junction.
abstract: Acetylcholinesterase (AChE) at the neuromuscular junction (NMJ) is anchored to the synaptic basal lamina via a triple helical collagen Q (ColQ). Congenital defects of ColQ cause endplate AChE deficiency and myasthenic syndrome. A single intravenous administration of adeno-associated virus serotype 8 (AAV8)-COLQ to Colq(-/-) mice recovered motor functions, synaptic transmission, as well as the morphology of the NMJ. ColQ-tailed AChE was specifically anchored to NMJ and its amount was restored to 89% of the wild type. We next characterized the molecular basis of this efficient recovery. We first confirmed that ColQ-tailed AChE can be specifically targeted to NMJ by an in vitro overlay assay in Colq(-/-) mice muscle sections. We then injected AAV1-COLQ-IRES-EGFP into the left tibialis anterior and detected AChE in noninjected limbs. Furthermore, the in vivo injection of recombinant ColQ-tailed AChE protein complex into the gluteus maximus muscle of Colq(-/-) mice led to accumulation of AChE in noninjected forelimbs. We demonstrated for the first time in vivo that the ColQ protein contains a tissue-targeting signal that is sufficient for anchoring itself to the NMJ. We propose that the protein-anchoring strategy is potentially applicable to a broad spectrum of diseases affecting extracellular matrix molecules.
authors.affiliation: Division of Neurogenetics, Center for Neurological Diseases and Cancer, Nagoya University Graduate School of Medicine, Nagoya, Japan.
authors.forename: Mikako
authors.initials: M
authors.lastname: Ito
authors.forename: Yumi
authors.initials: Y
authors.lastname: Suzuki
authors.forename: Takashi
authors.initials: T
authors.lastname: Okada
authors.forename: Takayasu
authors.initials: T
authors.lastname: Fukudome
authors.forename: Toshiro
authors.initials: T
authors.lastname: Yoshimura
authors.forename: Akio
authors.initials: A
authors.lastname: Masuda
authors.forename: Shin'ichi
authors.initials: S
authors.lastname: Takeda
authors.forename: Eric
authors.initials: E
authors.lastname: Krejci
authors.forename: Kinji
authors.initials: K
authors.lastname: Ohno
mesh_terms: D000110:Acetylcholinesterase / Q000235:genetics / Q000378:metabolism*; D000818:Animals; D003094:Collagen / Q000235:genetics* / Q000378:metabolism*; D000229:Dependovirus / Q000235:genetics; D015316:Genetic Therapy; D006801:Humans; D051379:Mice; D008822:Mice, Transgenic; D009124:Muscle Proteins / Q000235:genetics* / Q000378:metabolism*; D018482:Muscle, Skeletal / Q000378:metabolism; D009469:Neuromuscular Junction / Q000235:genetics / Q000378:metabolism*; D020511:Neuromuscular Junction Diseases / Q000235:genetics / Q000503:physiopathology / Q000628:therapy*; D009435:Synaptic Transmission
chemical_list: D009124:Muscle Proteins; D003094:Collagen; D000110:Acetylcholinesterase; C534672:Colq protein, mouse