FIGURE 2

Examples of time-lapse analysis. (A) As described in Figure 1A, single cell labeling of primary motor neurons was obtained for recycling endosomes (Rab11a-eGFP), combined with a membrane reporter (tagRFP-Caax) to identify cell type. Red boxes: Three cell compartments were imaged (2 Hz for 5 min), 1-axon initial segment, 2-mid-axonal segment, 3-axonal arbor segment. (B) Kymograms were generated from the time-lapses acquired (Kymograph tool, ImageJ) and a variety of transport metrics can be calculated manually (compiled in Excel, statistics in Graphpad Prism6). In this example, significant differences between neuronal segments are detected for the transport direction ratio (anterograde/retrograde), and retrograde run duration (n = 3 cells; AIS n = 47/54 anterograde/retrograde runs; mid-axon n = 85/82 anterograde/retrograde runs; arbor n = 63/86 anterograde/retrograde runs). ^∗∗p < 0.01, ^∗∗∗p < 0.001.