Interactions between neurofilament side arms may modulate axon caliber. To investigate this hypothesis, we derived transgenic mice expressing a fusion protein in which the carboxyl terminus of the high molecular weight neurofilament protein (NFH) was replaced by β-galactosidase. The transgene, regulated by NFH sequences, was expressed in projection neurons. However, the fusion protein remained in perikarya precipitating large filamentous aggregates. Axons were not invested with neurofilaments and developed only small calibers. Perikaryal aggregates, with similar structural features, are associated with neurodegenerative diseases, but these mice showed few ill effects and their neurons rarely degenerated. We conclude that an organized neurofilament cytoskeleton is required by axons to achieve large calibers but is not essential for neuronal function or extended survival.