Figure 5.

Retrograde axonal transport of targeted vectors. (a) Primary motor neurons plated in microfluidic chambers were incubated with either DiO labeled αp75^NTR or αThy1.1 or αCAR with TeNT-AF555 and anti-p75^NTR-AF647 for 2 hours at 37 °C before fixing and imaging by confocal microscopy. Shown are representative maximum projections of z-stack images where white circles show examples of colocalization of vectors with both markers and yellow circles show examples of colocalization with just TeNT-AF555 and green circles show non-associated vectors. Bars = 20 μm. (b) Shows quantification of colocalization (n = 3, 286 particles αp75^NTR, 160 particles αThy1.1, and 210 particles αCAR). (c) Primary motor neurons were incubated with either αp75^NTR, αCAR, or αThy1.1 DiO labeled vectors and anti-p75^NTR-AF647 for 60 minutes before time-lapse confocal imaging of a region of axon >200 μm from the axonal compartment. Representative image series of trafficking vector particles showing DiO vectors and p75^NTR (green and red in merged image, respectively). Asterisks indicate examples of stationery particle. Kymographs of the same series. Bars = 10 μm. CAR, coxsackievirus and adenovirus receptor.