As the spinal cord develops, many neural populations are established, each with characteristics critical for nervous system function. V2a interneurons are a key population of neurons involved in central pattern generation (CPG) and motor control within the spinal cord. Two major types of V2a neurons have been identified, which display marked shifts in their proportional abundance along the rostral-caudal axis of the mouse spinal cord, but the mechanisms that lead to this patterning remain to be molecularly described or controlled. To examine V2a neurons in vitro, I have established a lineage-traced human stem cell line based on expression of FOXN4 in V2a progenitors. My objective is to elucidate the molecular mechanisms that contribute to human V2a diversification and their impacts on neuron function. I hypothesize that axial identity, conferred by HOX transcription factors 1) regulate the generation of Type I and Type II V2a neurons though specific enhancers and 2) result in functional differences in CPG responsiveness and automaticity between axially distinct populations of V2a neurons.