A retrograde tracer strategy using True Blue to label the preganglionic parasympathetic innervation of the abdominal viscera.

A simple and reliable method for labeling the preganglionic neurons which innervate the abdominal viscera is described. Infusion of a True Blue (TB) suspension in several different concentrations directly into the peritoneal cavity consistently labeled the medullary vagal nuclei including the dorsal motor nucleus (DMN), nucleus ambiguus (NA), and retrofacial nucleus (RFN). Quantitative analysis of cell labeling after 30 microliters infusions of a large range of concentrations of TB (0.001-20.0%) showed that: (1) as little as 0.0075 mg of TB was sufficient to label DMN cells distinctly while only 0.075 mg (or larger) doses of TB were adequate for labeling cells of the NA and the RFN, and (2) doses of 3.0 mg (or greater) labeled the maximum number of cells in the DMN, NA, and RFN. Qualitative analysis suggested that medium range doses (0.075-0.75 mg) were optimal for discerning cell size and type throughout each of these nuclei.