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Fig. 8. Tyrosine phosphatase inhibition causes the accumulation of recycled
receptors in the peri-synaptic membrane. (A) Live adult mouse NMJ
previously saturated with BTX-biotin/strept-488 (green) at time 0 and viewed
at high detector gain 9 hours later. The muscle was continuously bathed in the
tyrosine phosphatase inhibitor PAO for 9 hours and then imaged. (B) The
junction was then incubated with strept-594 (red) to determine if PAO affects
AChR recycling. Perisynaptic areas were labeled (arrows), indicating that some
of the recycled receptors had become mistargeted. (C) Overlay of
pre-existing and recycled receptors (overlay images were adjusted for
brightness and contrast using Photoshop). (D-F) Similar results were
obtained using an alternative tyrosine phosphatase inhibitor, pervanadate.
Note that whereas recycled receptors (red) appear in the peri-synaptic area,
pre-existing receptors (green) remain restricted to the synapse. (G-I)
When the serine/threonine phosphatase inhibitor okadaic acid was used instead,
recycled receptors (red) were properly targeted to the synapse. Scale bars: 20
µm.
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