In an earlier study (Giesler et al., 1976), antidromically identified cells of origin of the rat spinothalamic tract (STT) were found to be widely distributed throughout the lumbar dorsal horn and intermediate gray zone (IGZ). Interestingly, it appeared that STT neurons located within the ventral dorsal horn and IGZ, which appeared to respond to stimuli delivered to subcutaneous tissue, tended to be activated only from midline thalamic structures. In contrast, STT neurons within nucleus proprius and the marginal zone, which responded to noxious and innocuous cutaneous stimuli, were activated from lateral thalamic structures. The present study has investigated this apparent dichotomy in thalamic afferents using the technique of retrograde transport of horseradish peroxidase (HRP). Single and multiple small injections of HRP were made in various thalamic regions. Tissue was reacted with diaminobenzidine (DAB) or o‐dianisidine (OD). Our data provide evidence that DAB histochemistry yields a more accurate reflection of the HRP concentrations surrounding an injection site which are necessary for visible retrograde transport. On the other hand, OD was found to be the more useful technique for examining labeled spinal cord neurons. With this method, 3 to 5.3 times more STT neurons were seen and far greater morphological detail was evident. Multiple HRP infusions which filled an entire hemi‐thalamus labeled large numbers of cells within the spinal extensions of the dorsal column nuclei, the lateral cervical nucleus, the ventral horns and IGZ bilaterally within C1 and C2, and the nucleus proprius and IGZ at all levels. STT neurons were also seen, but in lesser numbers within the marginal zone. Labeling in spinal segments below C3 was, with a single exception, exclusively contralateral. The lumbar enlargement was seen to contribute a greater number of STT neurons than the cervical enlargement. Injections into posterior medial thalamic structures produced bilateral labeling in the ventral horns and IGZ in upper cervical segments. Within the cervical and lumbar enlargements, labeled neurons were restricted to ventral dorsal horn and to IGZ contralaterally. With the exception of the ventral horn cells of C1 and C2 where a marked reduction in labeling occurred, more anterior medial thalamic injections produced labeling within the identical areas of the gray matter and in numbers comparable to those produced by more posterior medial thalamic injections. In contrast, injections into lateral thalamic areas yielded dense labeling within the caudal extensions of the dorsal column nuclei and lateral cervical nucleus. Labeled cells were seen in the marginal zone and nucleus proprius at all levels. Together with our earlier findings, these data suggest that the rat STT is composed of two components which vary in their cells of origin, their terminations and, very likely, their functions.