Distinct muscarinic acetylcholine receptor subtypes contribute to stability and growth, but not compensatory plasticity, of neuromuscular synapses.
Muscarinic acetylcholine receptors (mAChRs) modulate synaptic function, but whether they influence synaptic structure remains unknown. At neuromuscular junctions (NMJs), mAChRs have been implicated in compensatory sprouting of axon terminals in paralyzed or denervated muscles. Here we used pharmacological and genetic inhibition and localization studies of mAChR subtypes at mouse NMJs to demonstrate their roles in synaptic stability and growth but not in compensatory sprouting. M(2) mAChRs were present solely in motor neurons, whereas M(1), M(3), and M(5) mAChRs were associated with Schwann cells and/or muscle fibers. Blockade of all five mAChR subtypes with atropine evoked pronounced effects, including terminal sprouting, terminal withdrawal, and muscle fiber atrophy. In contrast, methoctramine, an M(2/4)-preferring antagonist, induced terminal sprouting and terminal withdrawal, but no muscle fiber atrophy. Consistent with this observation, M(2)(-/-) but no other mAChR mutant mice exhibited spontaneous sprouting accompanied by extensive loss of parental terminal arbors. Terminal sprouting, however, seemed not to be the causative defect because partial loss of terminal branches was common even in the M(2)(-/-) NMJs without sprouting. Moreover, compensatory sprouting after paralysis or partial denervation was normal in mice deficient in M(2) or other mAChR subtypes. We also found that many NMJs of M(5)(-/-) mice were exceptionally small and reduced in proportion to the size of parental muscle fibers. These findings show that axon terminals are unstable without M(2) and that muscle fiber growth is defective without M(5). Subtype-specific muscarinic signaling provides a novel means for coordinating activity-dependent development and maintenance of the tripartite synapse.
Wright
Megan C
MC
Department of Neurobiology, Drexel University College of Medicine, Philadelphia, Pennsylvania 19129, USA.
Potluri
Srilatha
S
Wang
Xueyong
X
Dentcheva
Eva
E
Gautam
Dinesh
D
Tessler
Alan
A
Wess
Jürgen
J
Rich
Mark M
MM
Son
Young-Jin
YJ
D000818:Animals; D001285:Atropine / Q000494:pharmacology; D003714:Denervation; D003959:Diamines / Q000494:pharmacology; D005260:Female; D020439:Growth Cones / Q000187:drug effects / Q000378:metabolism* / Q000648:ultrastructure; D051379:Mice; D008810:Mice, Inbred C57BL; D018345:Mice, Knockout; D009046:Motor Neurons / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D018727:Muscarinic Antagonists / Q000494:pharmacology; D018485:Muscle Fibers, Skeletal / Q000378:metabolism / Q000473:pathology; D018482:Muscle, Skeletal / Q000254:growth & development / Q000294:innervation / Q000378:metabolism; D009416:Nerve Regeneration / Q000187:drug effects / Q000235:genetics; D009469:Neuromuscular Junction / Q000187:drug effects / Q000235:genetics* / Q000378:metabolism*; D009473:Neuronal Plasticity / Q000187:drug effects / Q000235:genetics; D010243:Paralysis / Q000503:physiopathology; D017729:Presynaptic Terminals / Q000187:drug effects / Q000378:metabolism / Q000648:ultrastructure; D020033:Protein Isoforms / Q000187:drug effects / Q000235:genetics; D011976:Receptors, Muscarinic / Q000187:drug effects / Q000235:genetics*; D014855:Wallerian Degeneration / Q000139:chemically induced / Q000235:genetics / Q000378:metabolism
D003959:Diamines; D018727:Muscarinic Antagonists; D020033:Protein Isoforms; D011976:Receptors, Muscarinic; D001285:Atropine; C054938:methoctramine