Two techniques have been used to examine the organization of spinothalamic tract axons within the spinal cord of the rat. In the initial experiments, the thalamus was filled on one side with horseradish peroxidase (HRP) using a series of small injections. The injections were preceded by lesions of various areas of the ventral quadrant. These studies indicated that the cells of origin of STT axons ascending within the ventral funiculus (VF) are located primarily in the ventral-most areas of the dorsal horn and the intermediate gray zone. The cells of origin of STT axons projecting within the ventrolateral funiculus (VLF) are located not only deep within the gray matter but in addition within the dorsal-most two thirds of the dorsal horn, the area of the spinal cord gray matter shown in previous studies to contain the vast majority of cells with cutaneous tactile and nociceptive input. To examine these projections directly, rats received either a series of HRP injections that filled the thalamus on one side or a small injection into either medial or lateral thalamus. Examination of the labeled axons in horizontal sections through the cervical cord indicated that STT axons ascending to lateral thalamus do so in the VLF. In contrast, axons terminating in medial thalamus ascend in the VF. Additional experiments have shown that axons ascending to the lateral thalamus are distributed throughout the VLF at lumbar levels. Within the thoracic cord, lateral projecting SST axons are distributed throughout much of the VLF but are not found in close proximity to the ventral horn. At cervical levels all lateral-projecting STT axons have assumed a position on the lateral rim of the VLF. These and previously published data have demonstrated that the rat spinothalamic tract is composed of two components that differ in the distribution of their cells of origin, the area of the cord in which they ascend, and the thalamic nuclei in which they terminate.