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Fig. 2. Peripherin is present in growth cones. A living PC12 cell expressing
GFP-peripherin during the early stages of neurite outgrowth. This cell was
observed at
1-2 hours after plating in DM. Numerous GFP-peripherin
particles and squiggles can be seen within the growth cone. A-C represents a
single image from a time-lapse series. The image was captured by both
phase-contrast and fluorescence microscopy to show the relationships between
particles, squiggles, growth cones and filopodial extensions. The arrowheads
indicate the positions of peripherin particles, some of which can also be
detected with phase contrast. The particles and squiggles seen in the growth
cone region are motile (see Movie 1 at
jcs.biologists.org/supplemental).
Bar, 5 µm in A-C. PC12 cells were plated in DM for 2 hours and then
processed for platinum-replica immunogold TEM as described in Materials and
Methods using rabbit anti-peripherin and gold-conjugated secondary antibodies.
Ultrastructural observations (D-E) demonstrate that peripherin particles are
present within the actin-rich growth cones and filopodia. In more proximal
regions of growth cones, peripherin particles (indicated by clusters of 18 nm
gold), as well as peripherin squiggles (most probably represented by linear
arrays of gold; see arrows), are readily observed (F,G). E is a higher
magnification view of area in the red box in D; F is a higher magnification
view of the area in the green box in D; and G is a higher magnification view
of the area in the blue box in F. Bar, 100 nm in D,E,G; Bar, 500 nm in F.