Compartmentalization of acetylcholinesterase mRNA and enzyme at the vertebrate neuromuscular junction

Abstract

Acetylcholinesterase (AChE) is concentrated at the vertebrate neuromuscular synapse. To determine whether increased transcript levels could underlie this selective accumulation, we employed a quantitative reverse transcription polymerase chain reaction-based assay to determine mRNA copy number in samples as small as single neuromuscular junctions (NMJs) and a microassay to measure AChE enzyme activity at single synapses. Our results show that AChE mRNA is an intermediate transcript at NMJs, whereas in noninnervated regions of muscle fibers, AChE transcripts are either undetectable or rare. In contrast, alpha-actin transcript levels in the same samples are similar in junctional and extrajunctional regions. However, compared with AChE enzyme activity and alpha-actin mRNA levels, the levels of AChE transcripts at NMJs are highly variable. These results indicate that AChE mRNA and protein expression are compartmentalized at the vertebrate NMJ and provide a direct approach toward dissecting the molecular events leading from synaptic activation to plastic changes in gene expression at single vertebrate synapses.