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. 2001 Nov 15;21(22):8873–8885. doi: 10.1523/JNEUROSCI.21-22-08873.2001

Fig. 8.

Fig. 8.

Substance P expression strictly paralleled the expression of flg in cultured P-neurons, as illustrated by double-labeling immunofluorescence experiments. Left column shows examples of Trk-A (top row) or flg (other rows) immunoreactive neurons, and right column shows examples of substance P (SP) immunoreactive neurons. The rightand left panels in each row show the same view field. At E18 (top row), all P-neurons (arrowheads) expressed Trk-A(A) and none expressed SP immunoreactivity (B) (n = 12). A large number of round neurons were also immunoreactive for Trk-A(data not shown). At this stage there was no detectable immunolabeling for flg in P-neurons. At P2 (middle row), the P-neurons that were immunoreactive for flg(C) were also immunoreactive for SP (D) (n = 8). In contrast, those negative for flg were also negative for SP (data not shown). Only flg-positive P-neurons contained substance P. In non-P-neurons, coexpression of flg and SP was infrequent (∼7%). An example from P5 animals is shown inE and F. Arrowheadsindicate round neuron somata that coexpressed flg and SP, out of 29 round neurons present in the field. Arrowsindicate axons that showed SP, but not flg, immunoreactivity. In all cases, we used a rat monoclonal antibody against SP at a dilution of 1:20. Scale bars: A,B, 15 μm; C, D, 20 μm;E, F, 50 μm.