Lower back pain is an often chronic and debilitating condition that poses an economic burden of 100 billion annually in the United States alone. Based on recent projections made by the Global Burden of Disease Study, lower back pain is now predicted to be the leading cause of disability worldwide. Degenerative disc disease is a known contributor to lower back pain. Despite its high lifetime prevalence and economic burden, few effective therapies exist to reverse discogenic back pain. One critical obstacle to disc regenerative therapy, which aims to restore disc function by replacing damaged cells with healthy cells, is that the ontogeny of intervertebral disc cells and the mechanisms by which the discs are formed are not understood. Using the mouse model system we identified potential roles for ROBO-SLIT signaling during development of the intervertebral discs. ROBO-SLIT signaling is known to mediate organogenesis of the brain, kidneys, and foregut through a chemotactic mechanism. The role of ROBO-SLIT signaling during vertebral column and intervertebral disc morphogenesis is unknown. Using molecular approaches, we have determined that Robo and Slit mRNAs are expressed in complementary patterns during mouse disc development. We found that in the absence of ROBO1 and ROBO2 receptors the vertebral column forms with aberrantly shaped intervertebral discs that contain enlarged annulus fibrosus cells. Lineage tracing analysis using the Gt(ROSA)26Sortm4(ACTB-tdTomato,-EGFP)Luo (Rosa-mTmG) dual fluorescence reporter mouse showed that the disc phenotype was not caused by aberrant cell migration and/or cell mixing of nucleus pulposus an annulus fibrosis cells in the disc. Based on the dysregulated presence of Ihh and Col10a1 mRNA in the vertebral growth plates of double Robo mutant mice, we propose that the observed disc morphology may result from disruption of ROBO-mediated growth plate dynamics in vertebrae adjacent to the forming discs.