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Fig. 1. Recycled and pre-existing AChRs are removed at different rates from the
same synapse. The sternomastoid muscle was labeled first with
BTX-biotin/strept-488, then 3-4 days later with strept-594 to selectively
label recycled AChRs. (A) High resolution image of a NMJ branch showing
that recycled (red and yellow overlay) and pre-existing (retaining their
strept-488 after initial labeling; green and yellow overlay) AChRs are
intermingled in the postsynaptic membrane. (B) Preexisting and recycled
AChRs labeled with different fluorophores were assayed for fluorescence
intensities immediately after strept-594 labeling of recycled AChRs (d0), and
2 days later (d2). The total fluorescence intensity of each AChR pool was
normalized to 100% at initial imaging. Pseudo-color images provide a linear
representation of the density of AChRs. (C) Graph summarizing the
results obtained from all junctions by the approach shown in B. Each data
point represents the mean percentage of fluorescence intensity ±s.d.
Note that recycled receptors are removed significantly faster than
pre-existing receptors. Scale bars, 20 µm.
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