Quantum dot ex vivo labeling of neuromuscular synapses

Nicotinic receptors (nAchRs) are responsible for fast excitatory signaling by the neurotransmitter acetylcholine (Ach). They are present on the postsynaptic membrane at neuromuscular junctions (NMJs) and also at brain synapses. Alpha-bungarotoxin (alpha-BTX), a high-affinity nAchR antagonist, inhibits Ach binding and neurotransmission. Here we demonstrate biotinylated alpha-BTX, bound to native mouse diaphragm nAchRs, can be quantified and visualized ex vivo using streptavidin-conjugated quantum dots. This approach provides a novel methodology for the direct assessment of the presence and mobility of neurotransmitter receptors in native tissue.