title: Severely impaired neuromuscular synaptic transmission causes muscle weakness in the Cacna1a-mutant mouse rolling Nagoya.
abstract: The ataxic mouse rolling Nagoya (RN) carries a missense mutation in the Cacna1a gene, encoding the pore-forming subunit of neuronal Ca(v)2.1 (P/Q-type) Ca2+ channels. Besides being the predominant type of Ca(v) channel in the cerebellum, Ca(v)2.1 channels mediate acetylcholine (ACh) release at the peripheral neuromuscular junction (NMJ). Therefore, Ca(v)2.1 dysfunction induced by the RN mutation may disturb ACh release at the NMJ. The dysfunction may resemble the situation in Lambert-Eaton myasthenic syndrome (LEMS), in which autoantibodies target Ca(v)2.1 channels at NMJs, inducing severely reduced ACh release and resulting in muscle weakness. We tested neuromuscular function of RN mice and characterized transmitter release properties at their NMJs in diaphragm, soleus and flexor digitorum brevis muscles. Clinical muscle weakness and fatigue were demonstrated using repetitive nerve-stimulation electromyography, grip strength testing and an inverted grid hanging test. Muscle contraction experiments showed a compromised safety factor of neuromuscular transmission. In ex vivo electrophysiological experiments we found severely impaired ACh release. Compared to wild-type, RN NMJs had 50-75% lower nerve stimulation-evoked transmitter release, explaining the observed muscle weakness. Surprisingly, the reduction in evoked release was accompanied by an approximately 3-fold increase in spontaneous ACh release. This synaptic phenotype suggests a complex effect of the RN mutation on different functional Ca(v)2.1 channel parameters, presumably with a positive shift in activation potential as a prevailing feature. Taken together, our studies indicate that the gait abnormality of RN mice is due to a combination of ataxia and muscle weakness and that RN models aspects of the NMJ dysfunction in LEMS.
authors: Department of Neurology and Clinical Neurophysiology, Leiden University Medical Centre, Leiden, The Netherlands.
authors: Simon
authors: S
authors: Kaja
authors: Rob C G
authors: RC
authors: van de Ven
authors: J Gert
authors: JG
authors: van Dijk
authors: Jan J G M
authors: JJ
authors: Verschuuren
authors: Kiichi
authors: K
authors: Arahata
authors: Rune R
authors: RR
authors: Frants
authors: Michel D
authors: MD
authors: Ferrari
authors: Arn M J M
authors: AM
authors: van den Maagdenberg
authors: Jaap J
authors: JJ
authors: Plomp
mesh_terms: D000109:Acetylcholine / Q000378:metabolism; D000818:Animals; D001259:Ataxia / Q000503:physiopathology; D020864:Calcium Channels, N-Type; D020865:Calcium Channels, P-Type / Q000235:genetics / Q000378:metabolism*; D020907:Calcium Channels, Q-Type / Q000235:genetics / Q000378:metabolism*; D004576:Electromyography; D006801:Humans; D051379:Mice; D008817:Mice, Mutant Strains; D009119:Muscle Contraction / Q000502:physiology; D018763:Muscle Fatigue / Q000502:physiology; D018485:Muscle Fibers, Skeletal / Q000166:cytology / Q000378:metabolism; D018908:Muscle Weakness / Q000503:physiopathology*; D018482:Muscle, Skeletal / Q000166:cytology / Q000378:metabolism; D009469:Neuromuscular Junction / Q000502:physiology; D017354:Point Mutation; D009435:Synaptic Transmission / Q000502:physiology*
chemical_list: D000109:Acetylcholine; D020864:Calcium Channels, N-Type; D020865:Calcium Channels, P-Type; D020907:Calcium Channels, Q-Type