Figure 5.

Example of a quadruple labeling observed at the confocal level using a multi-track approach. In this image the following signals were simultaneously detected: CGRP (green); IB4 binding (red); CTb transported retrogradely from the parabrachial nucleus (blue); NK-1r (white). A fusiform neuron, double labeled with CTb and NK-1r, is innervated by CGRP-IR boutons (arrowhead) and IB4+ (arrow) boutons, which represent distinct populations. However, a small population of varicosities co-labeled for CGRP and IB4 (curved arrow) was detected. Scale bar (A-E) = 20 μm.