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Fig. 3. Heparan sulfate proteoglycans in C. elegans localize to the nerve ring, nerve cords and vulva. Whole worms were permeabilized, incubated with heparitinase and stained with anti-
-heparan sulfate antibody. (a) Staining shows that heparan sulfate proteoglycans are located at the nerve ring (arrowheads), ventral nerve cord (white arrows) and dorsal nerve cord (gray arrows). (b) Heparan sulfate proteoglycans are also located in the vulva as seen from a ventral view (gray arrowheads). The white arrows indicate the ventral nerve cord and the gray arrows the dorsal nerve cord. (c) The staining of L2-L3 larva showed the location of heparan sulfate proteoglycans at the nerve ring (white arrowhead) and at the ventral and dorsal nerve cords (white and gray arrows, respectively). (d) The nervous system of a transgenic strain expressing the pan-neural GFP was visualized. (e) Heparan sulfate proteoglycans as seen in panel a were also immunodetected in this reporter strain. The white arrows indicate ventral nerve cord. (f) Merged images of panels d and e. The insert shows an enlarged area, with details of the neural bodies (green) and their close localization to the heparan sulfate proteoglycans (red). Controls without heparitinase treatment and without primary antibody are shown in panels g and h respectively. All images are 3D reconstructions of image stacks, except for d, e and f, which correspond to a single optic slice. Bar, 20 µm.
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