Anti-agrin staining is absent at abandoned synaptic sites of frog neuromuscular junctions.
The neuromuscular junction is a plastic structure and is constantly undergoing changes as the nerve terminals that innervate the muscle fiber extend and retract their processes. In vivo observations on developing mouse neuromuscular junctions revealed that prior to the retraction of a nerve terminal the acetylcholine receptors (AChRs) under that nerve terminal disperse. Agrin is a protein released by nerve terminals that binds to synaptic basal lamina and directs the aggregation of AChRs and acetylcholinesterase (AChE) in and on the surface of the myotube. Thus, if the AChRs under a nerve terminal disperse, then the cellular signaling mechanism by which agrin maintains the aggregation of those AChRs, must have been disrupted. Two possibilities that could lead to the disruption of the agrin induced aggregation are that agrin is present at the synaptic basal lamina but is unable to direct the aggregation of AChRs, or that agrin has been removed from the synaptic basal lamina. Thus, if agrin were blocked, one would expect to see anti-agrin staining at abandoned synaptic sites; whereas if agrin were removed, anti-agrin staining would be absent at abandoned synaptic sites. We find that anti-agrin staining and alpha-bungarotoxin staining are absent at abandoned synaptic sites. Further, in vivo observations of retracting nerve terminals confirm that agrin is removed from the synaptic basal lamina within 7 days. Thus, while agrin will remain bound to synaptic basal lamina for months following denervation, it is removed within days following synaptic retraction.
Werle
M J
MJ
Department of Anatomy and Cell Biology, Kansas University Medical Center, Kansas City 66160-7400, USA. mwerle@kumc.edu
Sojka
A M
AM
D000110:Acetylcholinesterase / Q000032:analysis; D018171:Agrin / Q000032:analysis*; D000818:Animals; D000911:Antibodies, Monoclonal; D001001:Anura; D002038:Bungarotoxins / Q000032:analysis; D007150:Immunohistochemistry; D008297:Male; D009469:Neuromuscular Junction / Q000737:chemistry*; D011950:Receptors, Cholinergic / Q000378:metabolism*; D013194:Staining and Labeling; D013569:Synapses / Q000737:chemistry*
D018171:Agrin; D000911:Antibodies, Monoclonal; D002038:Bungarotoxins; D011950:Receptors, Cholinergic; D000110:Acetylcholinesterase