The actin cytoskeleton contributes directly or indirectly to nearly every aspect of neuronal development and function. This diversity of functions is often attributed to actin regulatory proteins, although how the composition of the actin cytoskeleton itself may influence its function is often overlooked. In neurons, the actin cytoskeleton is composed of two distinct isoforms, β- and γ-actin. Functions for β-actin have been investigated in axon guidance, synaptogenesis, and disease. Insight from loss-of-function in vivo studies has also revealed novel roles for β-actin in select brain structures and behaviors. Conversely, very little is known regarding functions of γ-actin in neurons. The dysregulation or mutation of both β- and γ-actin has been implicated in multiple human neurological disorders, however, demonstrating the critical importance of these still poorly understood proteins. This chapter highlights what is currently known regarding potential distinct functions for β- and γ-actin in neurons as well as the significant areas that remain unexplored.