Acetylcholinesterase dynamics at the neuromuscular junction of live animals.
At cholinergic synapses, acetylcholinesterase (AChE) is critical for ensuring normal synaptic transmission. However, little is known about how this enzyme is maintained and regulated in vivo. In this work, we demonstrate that the dissociation of fluorescently-tagged fasciculin 2 (a specific and selective peptide inhibitor of AChE) from AChE is extremely slow. This fluorescent probe was used to study the removal and insertion of AChE at individual synapses of living adult mice. After a one-time blockade of AChEs with fluorescent fasciculin 2, AChEs are removed from synapses initially at a faster rate (t(1/2) of approximately 3 days) and later at a slower rate (t(1/2) of approximately 12 days). Most of the removed AChEs are replaced by newly inserted AChEs over time. However, when AChEs are continuously blocked with fasciculin 2, the removal rate increases substantially (t(1/2) of approximately 12 h), and most of the lost AChEs are not replaced by newly inserted AChE. Furthermore, complete one-time inactivation of AChE activity significantly increases the removal of postsynaptic nicotinic acetylcholine receptors (AChRs). Finally, time lapse imaging reveals that synaptic AChEs and AChRs that are removed from synapses are co-localized in the same pool after being internalized. These results demonstrate a remarkable AChE dynamism and argue for a potential link between AChE function and postsynaptic receptor lifetime.
Krejci
Eric
E
Department of Molecular, Cellular, and Developmental Biology and Neuroscience Program, University of Michigan, 830 N. University Avenue, Ann Arbor, MI 48109, USA.
Martinez-Pena y Valenzuela
Isabel
I
Ameziane
Rafiqa
R
Akaaboune
Mohammed
M
D000110:Acetylcholinesterase / Q000378:metabolism / Q000502:physiology*; D000818:Animals; D019556:COS Cells; D002522:Chlorocebus aethiops; D004305:Dose-Response Relationship, Drug; D004546:Elapid Venoms / Q000378:metabolism; D005109:Extracellular Matrix / Q000378:metabolism; D005260:Female; D005456:Fluorescent Dyes / Q000494:pharmacology; D051379:Mice; D018613:Microscopy, Confocal; D008856:Microscopy, Fluorescence / Q000379:methods*; D009469:Neuromuscular Junction / Q000378:metabolism*; D011978:Receptors, Nicotinic / Q000378:metabolism; D013569:Synapses / Q000378:metabolism; D013997:Time Factors; D014118:Toxins, Biological / Q000737:chemistry
D004546:Elapid Venoms; D005456:Fluorescent Dyes; D011978:Receptors, Nicotinic; D014118:Toxins, Biological; C044937:fasciculin; D000110:Acetylcholinesterase