We report a specific tracing technique for studying projections of noradrenergic neurons that contain other transmitters. Autoradiography after retrograde axonal transport of tritiated-noradrenaline ([3H]NA) was combined with immunocytochemical detection of endogenous NA or neuropeptide Y (NPY). The specificity of [3H]NA retrograde labeling was dependent on the concentration of [3H]NA injected at the terminal region. At 2 x 10(-3) M, the specificity of [3H]NA retrograde labeling was confirmed by immunodetection of endogenous NA in radiolabeled cell bodies of the locus ceruleus. Combination of autoradiography of [3H]NA retrograde labeling with immunodetection of NPY demonstrated that a some scattered locus ceruleus noradrenergic neurons (about 26%) projecting to the olfactory bulb do contain NPY.
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