Lead-induced attenuation in the aggregation of acetylcholine receptors during the neuromuscular junction formation

Shun Sheng Chen, Chuang Hao Lin, Tsan Ju Chen*

*Corresponding author for this work

Research output: Contribution to journalJournal Article peer-review

10 Scopus citations

Abstract

Lead (Pb2+) toxicity is more common in children and is associated with cognitive deficits, which may reflect lead-induced changes in central synaptic development and function. Aside from neurotoxicity, lead exposure may also impact mature neuromuscular junction (NMJ) and cause muscle weakness. NMJ is known as a peripheral cholinergic synapse and its signaling cascades responsible for development are similar to those for the central synapses. However, the effect of lead exposure on the formation of NMJ in mammals is unclear. In the present study, a NG108-15/C2C12 coculture model was used to measure the acetylcholine receptor (AChR) aggregates formed on the myotubes which was an early hallmark for the NMJ formation. AChR aggregates were identified by α-bungarotoxin under fluorescent microscope. Single dose of lead acetate with final concentrations at 10-3, 10-1, or 10 μM was applied to dishes at the beginning of coculturing. Following 3-day exposure, although NG108-15 cells could extend long neurites to nearby myotubes, obvious dose-dependent attenuation in AChR aggregation was shown. The averaged area of an AChR aggregate, the averaged number of AChR aggregates per myotube, and the total area of AChR aggregates per myotube were all significantly decreased. In addition, the distribution percentages of various sizes of AChR aggregates showed that almost half of the AChR aggregates were formed with a size of 2-5 μm2 regardless of lead exposure. After treating 10 μM of lead acetate, significantly more AChR aggregates ranged from 2 to 20 μm2 were formed and significantly less AChR aggregates larger than 20 μm2 were formed. These results indicated that lead exposure reduced the extent of AChR aggregation concerning both the size and number of AChR aggregates and large AChR aggregates could hardly be formed after acute high-level lead exposure. No significant change was found in the total amount of AChRs on the myotubes after lead exposure, which indicated that the attenuation of AChR aggregation was not caused by reducing the synthesis of AChRs but by remaining dispersed pattern of AChRs on the myotubes. These data suggest that lead exposure exerts detrimental effects on the formation of NMJ.

Original languageEnglish
Pages (from-to)89-99
Number of pages11
JournalToxicology Letters
Volume159
Issue number1
DOIs
StatePublished - 15 10 2005

Keywords

  • Acetylcholine receptor aggregation
  • Lead toxicity
  • Neuromuscular junction

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