Acetylcholinesterase mobility and stability at the neuromuscular junction of living mice.
Acetylcholinesterase (AChE) is an enzyme that terminates acetylcholine neurotransmitter function at the synaptic cleft of cholinergic synapses. However, the mechanism by which AChE number and density are maintained at the synaptic cleft is poorly understood. In this work, we used fluorescence recovery after photobleaching, photo-unbinding, and quantitative fluorescence imaging to investigate the surface mobility and stability of AChE at the adult innervated neuromuscular junction of living mice. In wild-type synapses, we found that nonsynaptic (perisynaptic and extrasynaptic) AChEs are mobile and gradually recruited into synaptic sites and that most of the trapped AChEs come from the perijunctional pool. Selective labeling of a subset of synaptic AChEs within the synapse by using sequential unbinding and relabeling with different colors of streptavidin followed by time-lapse imaging showed that synaptic AChEs are nearly immobile. At neuromuscular junctions of mice deficient in alpha-dystrobrevin, a component of the dystrophin glycoprotein complex, we found that the density and distribution of synaptic AChEs are profoundly altered and that the loss rate of AChE significantly increased. These results demonstrate that nonsynaptic AChEs are mobile, whereas synaptic AChEs are more stable, and that alpha-dystrobrevin is important for controlling the density and stability of AChEs at neuromuscular synapses.
Martinez-Pena y Valenzuela
Isabel
I
Department of Molecular, Cellular, and Developmental Biology, and Neuroscience Program, University of Michigan, Ann Arbor, MI 48109, USA.
Akaaboune
Mohammed
M
D000110:Acetylcholinesterase / Q000378:metabolism*; D000818:Animals; D049029:Dystrophin-Associated Proteins / Q000172:deficiency; D004795:Enzyme Stability; D005260:Female; D051379:Mice; D009469:Neuromuscular Junction / Q000166:cytology / Q000201:enzymology*; D021381:Protein Transport
D049029:Dystrophin-Associated Proteins; C098885:dystrobrevin; D000110:Acetylcholinesterase