Transplantation of quail collagen-tailed acetylcholinesterase molecules onto the frog neuromuscular synapse

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Abstract

The highly organized pattern of acetylcholinesterase (ACHE) molecules attached to the basal lamina of the neuromuscular junction (NMJ) suggests the existence of specific binding sites for their precise localization. To test this hypothesis we immunoaffinity purified quail globular and collagen- tailed AChE forms and determined their ability to attach to frog NMJs which had been pretreated with high-salt detergent buffers. The NMJs were visualized by labeling acetylcholine receptors (AChRs) with TRITC-α- bungarotoxin and AChE by indirect immunofluorescence; there was excellent correspondence (>97%) between the distribution of frog AChRs and AChE. Binding of the exogenous quail AChE was determined using a species-specific monoclonal antibody. When frog neuromuscular junctions were incubated with the globular G4/G2 quail AChE forms, there was no detectable binding above background levels, whereas when similar preparations were incubated with the collagen-tailed A12 AChE form >80% of the frog synaptic sites were also immunolabeled for quail AChE attached. Binding of the A12 quail AChE was blocked by heparin, yet could not be removed with high salt buffer containing detergent once attached. Similar results were obtained using empty myofiber basal lamina sheaths produced by mechanical or freeze-thaw damage. These experiments show that specific binding sites exist for collagen-tailed AChE molecules on the synaptic basal lamina of the vertebrate NMJ and suggest that these binding sites comprise a 'molecular parking lot' in which the AChE molecules can be released, retained, and turned over.

Original languageEnglish
Pages (from-to)367-374
Number of pages8
JournalJournal of Cell Biology
Volume136
Issue number2
DOIs
StatePublished - Feb 15 1997

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Quail
Acetylcholinesterase
Anura
Synapses
Collagen
Transplantation
Neuromuscular Junction
Basement Membrane
compound A 12
Binding Sites
Cholinergic Receptors
Detergents
Buffers
Salts
Bungarotoxins
Indirect Fluorescent Antibody Technique
Vertebrates
Heparin
Monoclonal Antibodies

ASJC Scopus subject areas

  • Cell Biology

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Transplantation of quail collagen-tailed acetylcholinesterase molecules onto the frog neuromuscular synapse. / Rotundo, Richard L; Rossi, Susana G; Anglister, Lili.

In: Journal of Cell Biology, Vol. 136, No. 2, 15.02.1997, p. 367-374.

Research output: Contribution to journalArticle

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