Many physical systems break symmetry in their evolution. Biophysical systems, such as cells, developing organisms, and even entire populations, are no exception. Developing neurons represent a striking example of a biophysical system that breaks symmetry: neurons cultured in vitro begin as cell bodies with several tendrils (“neurites”) growing outward. A few days later, these same neurons invariably have the same new morphology: exactly one of the neurites (the “axon”) has grown hundreds of microns in length, while the others (the “dendrites”) are much shorter and are more branched. Previous work has shown that any of the neurites can become the axon, and so neurons must break symmetry during their development. The mechanisms underlying neuronal symmetry breaking and axon specification have recently attracted attention, with multiple groups proposing biophysical models to explain the phenomena. In this thesis, we perform the first analytical comparisons of these models by conducting multiple phenotypic and morphological studies of neurite growth in developing neurons. Studying neurite dynamics is technically challenging because neurites have unpredictable morphologies. In Chapter 4, we study neurite competition and neuronal symmetry breaking in hundreds of neurons by optically patterning micron-wide stripes to which the neurons adhere, and on which they grow exactly two neurites. We then use our measurements to test the accuracy of the models in the simple case when a neuron has exactly two neurites. In Chapter 5, we no longer constrain neuronal morphology. One characteristic of symmetry breaking systems is how the system’s complexity affects the symmetry breaking. We find that a majority of the models predict that neurons with more neurites break symmetry much slower than neurons with fewer neurites. Experimentally, we find that neurons with different neurite counts break symmetry at the same rate, consistent with previous observations. We then determine why the models disagree in their predictions, and rectify the models using our own experimental data. In particular, we find that neurons with higher neurite counts have higher concentrations of key proteins involved in symmetry breaking, so that neurons, regardless of neurite count, can break symmetry at the same rate. / Physics