Effects of Chronic Lead Exposure on the Neuromuscular Junction in Drosophila Larvae

Abstract

Long term or chronic exposure to lead is associated with cognitive and other deficits in humans, which may reflect lead-induced changes in synaptic development and function. We believe that Drosophila has great potential as a model system for studying such changes. To test this, we compared the structure of single, identified synapses between identified axons (axons 1 and 2) and muscle fibers (fibers 6 and 7) in untreated 3rd instar larvae, and in larvae reared on medium made with 100 μM lead acetate in distilled water. We used three approaches to examine the motor terminals on muscle fibers 6 and 7 in segment 2: (1) all terminals were stained with an antibody to HRP; (2) only the terminals of axon 1 were stained by injecting biotinylated Lucifer yellow into it; and (3) the regions of the terminal containing synaptic vesicles were stained with an antibody to synaptotagmin, which provides an estimate of “synaptic” terminal area. Lead burdens were determined by inductively coupled plasma mass spectrometry; hemolymph lead levels at the neuromuscular junction were likely to be micromolar. We observed that lead exposure did not significantly affect the average terminal area or the average muscle fiber area, but did significantly affect the uniformity of the matching between muscle area and motor terminal size that normally occurs during development. There was a significant positive correlation between motor terminal size and muscle area in control, but not in lead-exposed larvae. The sensitivity of Drosophila larval synaptic development to lead opens the way to using the powerful genetic and molecular tools available for this system to study the underlying mechanisms of this sensitivity. We would hope that from such an understanding may come strategies for dealing with lead-induced deficits in children.